TW202335674A - Lipid conjugation for targeting astrocytes of the central nervous system - Google Patents

Abstract

Oligonucleotide conjugates are provided herein that inhibit or reduce expression of target genes in the astrocytes of the central nervous system. Also provided are compositions including the same and uses thereof, particularly uses relating to treating diseases, disorders and/or conditions associated with expression of an astrocyte target gene in the CNS.

Description

Lipid conjugates targeting stellate cells in the central nervous system

The present disclosure relates to oligonucleotides linked to lipid moieties that can be used to inhibit target genes in stellate cells of the central nervous system. Specifically, the present disclosure relates to oligonucleotide-lipid conjugates, methods of making them, their chemical configuration, and modulation using nucleic acids and oligonucleotides according to the conjugation described herein. A method of inhibiting (e.g., reducing) the expression of a target gene in stellate cells of the central nervous system (hereinafter referred to as "CNS") (e.g., a tissue, or region of the CNS). The present disclosure also provides pharmaceutically acceptable compositions containing the combinations of the present specification and methods of using the compositions to treat various diseases or conditions. [ Cross-reference to related applications ] This application claims priority to U.S. Provisional Patent Application No. 63/276,406, filed on November 5, 2021, which is incorporated herein by reference in its entirety. [ Reference Electronic Sequence Listing ] The contents of the electronic sequence listing (DICN_013_001TW_SeqList_ST26.xml; size: 344,190 bits; and creation date: November 2, 2022) are incorporated herein by reference in their entirety.

Modulating gene expression through modified nucleic acids shows great potential both as a laboratory research tool and as a clinical treatment method. Several types of oligonucleotide or nucleic acid-based treatments are already in clinical research, including antisense oligonucleotides (ASO), short interfering RNA (siRNA), and double-stranded nucleic acids. nucleic acid (dsNA), aptamers, ribozymes, exon-skipping and splice-altering oligonucleotides, immunomodulatory oligonucleotides, mRNA, and CRISPR. Chemical modification of relevant molecules to function in various tissues, organs, and/or cell types in overcoming the challenges of oligonucleotide therapeutics, including improving nuclease stability, RNA binding affinity, and pharmacokinetics Sex plays a key role. Over the past three decades, various chemical modification strategies for oligonucleotides (including modifications of sugars, nucleobases, and phosphodiester backbones) have been developed to improve and optimize performance and therapeutic efficacy (Deleavey and Darma, CHEM. BIOL.2012, 19(8):937-54; Wan and Seth, J. MED.CHEM.2016, 59(21): 9645-67; and Egli and Manoharan, ACC. CHEM.RES.2019, 54(4 ):1036-47). Therapeutic gene silencing mediated by RNAi oligonucleotide-based therapies including siRNA and double-stranded nucleic acids (dsNA) offers the potential to significantly expand the druggable target space and provide new opportunities for therapeutics This offers possibilities for orphan diseases that may not be treatable by other drug modalities (e.g., antibodies and/or small molecules). RNAi oligonucleotide-based therapies that inhibit or reduce the expression of specific target genes in the liver have been developed and are currently in clinical use (Sehgal et al., (2013) JOURNAL OF HEPATOLOGY 59:1354-59). There are still technical obstacles to the development and clinical use of RNAi oligonucleotides in extrahepatic cells, tissues, and organs (eg, CNS). Therapeutic gene silencing in the CNS mediated by RNAi oligonucleotide-based therapies is of particular interest for the treatment of neurological diseases (Boudreau & Davidson (2010) BRAIN RESEARCH 1338:112-21). Accordingly, there is a continuing need in the art to successfully develop novel and effective RNAi oligonucleotides to modulate the expression of target genes in extrahepatic cells, tissues, and/or organs (eg, CNS). This is complicated by the variable nature of extrahepatic cell types and considerations of circulation patterns and cell membrane components (such as receptor types).

The mammalian CNS is a complex system of cells, fluids, and chemicals that interact together to achieve a wide variety of functions, including movement, navigation, cognition, speech, vision, and emotion. Unfortunately, there are a variety of diseases and conditions of the CNS (eg, neurological disorders) that are known to affect or disrupt some or all of these functions. In general, treatment of CNS diseases and disorders is limited to small molecule drugs, antibodies, and/or adaptive or behavioral therapies. There is an ongoing need to develop treatments for diseases and conditions of the CNS associated with inappropriate gene expression. The present disclosure is based at least in part on the discovery that lipid-conjugated RNAi oligonucleotides effectively reduce target gene expression in CNS stellate cells. Exemplary lipid-conjugated RNAi oligonucleotides provided herein have been shown to reduce target gene expression of stellate cell-specific mRNA in the CNS after a single administration. Furthermore, the exemplary lipid-conjugated RNAi oligonucleotides provided herein have demonstrated pharmacological activity in multiple regions throughout the CNS, including hard-to-reach regions such as the hippocampus and frontal cortex. Without being bound by theory, hydrophobic moieties (eg, lipids) facilitate the delivery and distribution of lipid-bound RNAi oligonucleotides into the CNS, thereby increasing the efficacy and persistence of gene attenuation in stellate cells. Accordingly, the present disclosure provides methods of treating a disease or disorder by modulating the expression of stellate cell genes in the CNS using the lipid-conjugated RNAi oligonucleotides described herein, and pharmaceutically acceptable compositions thereof. The present disclosure further provides methods of using lipid-conjugated RNAi oligonucleotides in the manufacture of agents for treating diseases or disorders by modulating the expression of stellate cell genes in the CNS. Accordingly, in some aspects, the present disclosure provides double-stranded oligonucleotides comprising an antisense strand that is 15 to 30 nucleotides long and a sense strand that is 15 to 50 nucleotides long, wherein the antisense strand and The sense strand forms a double-stranded region of 15 to 30 base pairs, wherein the antisense strand contains a region complementary to the stellate cell mRNA target sequence, and wherein the sense strand contains at least one lipid moiety that binds to the nucleotides of the sense strand. In some aspects, the lipid moiety is selected from In some aspects, the lipid moiety is a hydrocarbon chain. In some aspects, the hydrocarbon chain is a C8 to C30 hydrocarbon chain. In some aspects, the hydrocarbon chain is a C16 hydrocarbon chain. In some forms, the C16 hydrocarbon chain consists of represented. In any of the foregoing or related aspects, the lipid moiety is bound to the 2' carbon of the ribose ring of the nucleotide. In any of the foregoing or related aspects, the oligonucleotide is blunt-ended. In some aspects, the oligonucleotide is blunt-ended at the 3' end of the oligonucleotide. In some aspects, the oligonucleotide contains blunt ends. In some aspects, the blunt end includes the 3' end of the positive strand. In some forms, the sense strand is 20 to 22 nucleotides. In some aspects, at least one lipid moiety binds to a nucleotide at position 1, position 4, position 8, position 12, position 13, position 18, or position 20 of the sense strand, wherein the position is from 5' to 3' number. In some aspects, the stellate cell mRNA target is expressed in the spinal cord, wherein the at least one lipid moiety is associated with position 1, position 4, position 8, position 12, position 13, position 18, or position 20 of the sense strand The nucleotides are bound at and the positions are numbered from 5' to 3'. In other aspects, the stellate cell mRNA target is expressed in the medulla oblongata, wherein the at least one lipid moiety is associated with position 1, position 4, position 8, position 12, position 13, position 18, or position 20 of the sense strand The nucleotides are bound at and the positions are numbered from 5' to 3'. In a further aspect, the stellate cell mRNA target is expressed in the cerebellum, wherein the at least one lipid moiety binds to a nucleotide at position 4, position 12, position 13, position 18, or position 20 of the sense strand , and the positions are numbered from 5' to 3'. In some aspects, the stellate cell mRNA target is expressed in the hypothalamus, wherein the at least one lipid moiety is associated with position 1, position 4, position 12, position 13, position 18, or position 20 of the sense strand. Nucleotides are bound, and positions therein are numbered from 5' to 3'. In other aspects, the stellate cell mRNA target is expressed in the frontal cortex, wherein the at least one lipid moiety binds to nucleotide at position 4 of the sense strand, and wherein the position is from 5' to 3' number. In any of the foregoing or related aspects, the sense strand includes a backbone-loop at the 3' end, wherein the backbone-loop includes a nucleotide sequence represented by the formula: 5'-S1-L-S2-3', Where S1 is complementary to S2, and L forms a loop between S1 and S2. In some forms, the sense strand is 36 to 38 nucleotides long. In some aspects, the stellate cell mRNA target is expressed in the spinal cord, wherein the at least one lipid moiety is associated with position 1, position 4, position 8, position 12, position 13, position 18, position 20, The nucleotide at position 23, position 28, position 29, or position 30 binds, and the positions are numbered from 5' to 3'. In other aspects, the stellate cell mRNA target is expressed in the medulla oblongata, wherein the at least one lipid moiety is associated with position 1, position 4, position 18, position 20, position 23, position 28, position 29, Or the nucleotide at position 30 binds, and the positions are numbered from 5' to 3'. In a further aspect, the stellate cell mRNA target is expressed in the cerebellum, wherein the at least one lipid moiety binds to a nucleotide at position 1, position 4, position 23, position 28, or position 29 of the sense strand , and the positions are numbered from 5' to 3'. In some aspects, the stellate cell mRNA target is expressed in the hypothalamus, wherein the at least one lipid moiety is associated with position 1, position 4, position 12, position 13, position 18, position 20, position 23 of the sense strand , the nucleotide at position 28, position 29, or position 30 binds, and the positions are numbered from 5' to 3'. In other aspects, the stellate cell mRNA target is expressed in the frontal cortex, wherein the at least one lipid moiety binds to nucleotide at position 23 of the sense strand, and wherein the position is from 5' to 3' number. In any of the foregoing or related aspects, the antisense strand is 22 to 24 nucleotides. In some aspects, the double-stranded region is 20 to 22 base pairs. In any of the foregoing or related aspects, the antisense strand comprises 1 to 4 nucleotide overhangs at the 3' end. In some aspects, the overhangs comprise purine nucleotides. In some aspects, the overhang sequence is 2 nucleotides long. In some aspects, the overhang is selected from AA, GG, AG, and GA. In some aspects, the overhang is GG or AA. In some versions, the overhang is GG. In any of the foregoing or related aspects, the region of complementarity is complementary to at least 15 contiguous nucleotides of the stellate cell mRNA target sequence. In some aspects, the complementary region is complementary to 19 contiguous nucleotides of the stellate cell mRNA target sequence. In some aspects, the complementary region is completely complementary to the stellate cell mRNA target sequence. In some aspects, the complementary region is partially complementary to the stellate cell mRNA target sequence. In some aspects, the complementary region contains no more than four mismatches to the stellate cell mRNA target sequence. In some aspects, the complementary region contains up to four mismatches to the stellate cell mRNA target sequence. In any of the preceding or related aspects, the oligonucleotide comprises at least one modified nucleotide. In some aspects, the modified nucleotides comprise 2'-modifications. In some aspects, in addition to the nucleotides of the sense strand bound to the at least one lipid moiety, each of the nucleotides of the sense strand and the antisense strand comprise 2'-modifications. In some aspects, the 2'-modification is selected from 2'-aminoethyl, 2'-fluoro, 2'-O-methyl, 2'-O-methoxyethyl, and 2'-des Modification of oxy-2'-fluoro-β-d-arabinose nucleic acid. In some aspects, the sense strand consists of about 10 to 20%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% contained 2'-fluoro modifications. In some forms, the antisense strand contains approximately 25 to 35%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, or 35% containing 2'-fluoro modifications. In some aspects, the oligonucleotide is about 25 to 35%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34% , or 35% contains 2'-fluorine modification. In some aspects, the sense strand includes 20 nucleotides from 5' to 3' positions 1 to 20, wherein each of positions 8 to 11 includes a 2'-fluoro modification. In some aspects, the sense strand includes 20 nucleotides from 5' to 3' positions 1 to 20, wherein each of positions 9 to 11 includes a 2'-fluoro modification. In some aspects, the sense strand includes 36 nucleotides from 5' to 3' positions 1 to 36, wherein each of positions 8 to 11 includes a 2'-fluoro modification. In some aspects, the sense strand includes 36 nucleotides from 5' to 3' positions 1 to 36, wherein each of positions 9 to 11 includes a 2'-fluoro modification. In some aspects, the antisense strand includes 22 nucleotides having positions 1 to 22 from 5' to 3', and wherein each of positions 2, 3, 4, 5, 7, 10, and 14 includes 2' -Fluorine modification. In some aspects, except for the nucleotide of the sense strand bound to the at least one lipid moiety, the remaining nucleotides comprise a 2'-O-methyl modification. In any of the preceding or related aspects, the oligonucleotide comprises at least one modified internucleotide linkage. In some aspects, at least one modified internucleotide linkage is linked to a phosphorothioate linkage. In some aspects, the antisense strand includes (i) between positions 1 and 2, and between positions 2 and 3; or (ii) between positions 1 and 2, between positions 2 and 3, and a phosphorothioate linkage between positions 3 and 4, where positions are numbered 1 to 4 from 5' to 3'. In some aspects, the antisense strand is 22 nucleotides long, and wherein the antisense strand includes a phosphorothioate linkage between positions 20 and 21 and between positions 21 and 22, wherein positions from 5' to 3' are numbered 1 to 22. In some aspects, the sense strand includes a phosphorothioate linkage between positions 1 and 2, with positions numbered 1 to 2 from 5' to 3'. In some aspects, the sense strand is 20 nucleotides long, and wherein the sense strand includes a phosphorothioate linkage between positions 18 and 19, and between positions 19 and 20, wherein positions 5' to 3' are numbered from 1 to 22. In any of the foregoing or related aspects, the antisense strand comprises a phosphorylated nucleotide at the 5' end, wherein the phosphorylated nucleotide is selected from uridine and adenosine. In some aspects, the phosphorylated nucleotide is uridine. In some aspects, the 4'-carbon of the sugar of the 5'-nucleotide of the antisense strand contains a phosphate analog. In some aspects, the phosphate analog is oxymethylphosphonate, vinylphosphonate, or malonylphosphonate. In any of the foregoing or related aspects, the region of complementarity is completely complementary to the stellate cell mRNA target sequence at nucleotide positions 2 to 8 of the antisense strand, where the nucleotide positions are numbered from 5' to 3'. In some aspects, the region of complementarity is completely complementary to the stellate cell mRNA target sequence at nucleotide positions 2 to 11 of the antisense strand, where the nucleotide positions are numbered from 5' to 3'. In any of the foregoing or related aspects, the oligonucleotide is a Dicer substrate. In some forms, the oligonucleotide is a Dicer substrate that, upon processing by endogenous Dicer, produces a 19 to 21 nucleotide long double strand that is capable of reducing stellate cell mRNA expression in mammalian cells. nucleic acids. In any of the foregoing or related aspects, the stellate cell mRNA target sequence is located in a region of the central nervous system (CNS). In some aspects, the region of the CNS is selected from the group consisting of spinal cord, lumbar spinal cord, thoracic spinal cord, cervical spinal cord, medulla oblongata, hippocampus, cerebellum, hypothalamus, frontal cortex, and combinations thereof. In any of the foregoing or related aspects, the oligonucleotide reduces the expression of target mRNA in stellate cells or stellate cell populations in vitro and/or in vivo. In some aspects, the present disclosure provides pharmaceutical compositions comprising an oligonucleotide described herein, and a pharmaceutically acceptable carrier, delivery agent, or excipient. In other aspects, the present disclosure provides methods of treating an individual suffering from a disease, disorder, or condition associated with expression of stellate cell mRNA, the method comprising administering to the individual a therapeutically effective amount of an oligonucleotide described herein. glycosides or pharmaceutical compositions to treat an individual. In a further aspect, the present disclosure provides methods of delivering oligonucleotides to stellate cells or populations of stellate cells in an individual, the method comprising administering to the individual a pharmaceutical composition described herein. In some forms, stellate cells or populations of stellate cells are located in regions of the CNS. In some aspects, the region of the CNS is selected from the group consisting of spinal cord, lumbar spinal cord, thoracic spinal cord, cervical spinal cord, medulla oblongata, hippocampus, cerebellum, hypothalamus, frontal cortex, and combinations thereof. In other aspects, the present disclosure provides methods of reducing the expression of stellate cell mRNA in a cell, cell population, or individual, the method comprising the steps of: i. contacting the cell or cell population with an oligonucleotide described herein or contacting the pharmaceutical composition, optionally wherein the cells or cell populations are stellate cells or populations of stellate cells; or ii. administering to the individual an oligonucleotide or pharmaceutical composition described herein. In some aspects, reducing the expression of stellate cell mRNA includes reducing the amount or level of mRNA, the amount or level of protein, or both. In some aspects, the individual suffers from a disease, disorder, or condition associated with expression of stellate cell mRNA. In some aspects, cells or cell populations are located in regions of the CNS. In some aspects, the region of the CNS is selected from the group consisting of spinal cord, lumbar spinal cord, thoracic spinal cord, cervical spinal cord, medulla oblongata, hippocampus, cerebellum, hypothalamus, frontal cortex, and combinations thereof. In any of the foregoing or related aspects, the method includes administering via intrathecal administration. In some aspects, the present disclosure provides methods of reducing the expression of a target mRNA expressed in stellate cells in tissues of the CNS of an individual, comprising administering to the individual a double-stranded oligonucleotide, the double-stranded oligonucleotide The acid contains an antisense strand of 15 to 30 nucleotides long and a sense strand of 15 to 50 nucleotides long, wherein the antisense strand and the sense strand form a double-stranded region of 15 to 30 base pairs, in which the antisense strand The strand contains a region complementary to the target sequence in the target mRNA, and wherein the sense strand contains at least one lipid moiety that binds to the nucleotides of the sense strand. In some aspects, the lipid moiety is a C16 hydrocarbon. In any of the preceding or related aspects of the methods described herein, the oligonucleotide is blunt-ended at the 3' end of the oligonucleotide. In some aspects, the oligonucleotide contains blunt ends. In some aspects, the blunt end includes the 3' end of the positive strand. In some forms, the sense strand is 22 to 24 nucleotides in length. In some aspects, the tissue is spinal cord, wherein the at least one lipid moiety binds to a nucleotide at position 1, position 4, position 8, position 12, position 13, position 18, or position 20 of the sense strand, And the positions are numbered from 5' to 3'. In other aspects, the tissue is the medulla oblongata, wherein the at least one lipid moiety binds to a nucleotide at position 1, position 4, position 8, position 12, position 13, position 18, or position 20 of the sense strand, And the positions are numbered from 5' to 3'. In a further aspect, the tissue is cerebellum, wherein the at least one lipid moiety binds to a nucleotide at position 4, position 12, position 13, position 18, or position 20 of the sense strand, and wherein the position is from 5 'To 3' number. In some aspects, the tissue is the hypothalamus, wherein the at least one lipid moiety binds to a nucleotide at position 1, position 4, position 12, position 13, position 18, or position 20 of the sense strand, and The positions are numbered from 5' to 3'. In other aspects, the tissue is frontal cortex, wherein the at least one lipid moiety binds to a nucleotide at position 4 of the sense strand, and wherein the positions are numbered from 5' to 3'. In any of the foregoing or related aspects of the methods described herein, the righteous strand includes a backbone-loop at the 3' end, wherein the backbone-loop includes the formula: 5'-S1-L-S2-3' Represents a nucleotide sequence in which S1 is complementary to S2 and in which L forms a loop between S1 and S2. In some forms, the sense strand is 36 to 38 nucleotides long. In some aspects, the tissue is the spinal cord, wherein the at least one lipid moiety is associated with position 1, position 4, position 8, position 12, position 13, position 18, position 20, position 23, position 28, position 29, or the nucleotide at position 30 is bound, and the positions are numbered from 5' to 3'. In other aspects, the tissue is the medulla oblongata, wherein the at least one lipid moiety is associated with a nucleoside at position 1, position 4, position 18, position 20, position 23, position 28, position 29, or position 30 of the sense strand Acid binds and the positions are numbered from 5' to 3'. In a further aspect, the tissue is cerebellum, wherein the at least one lipid moiety binds to a nucleotide at position 1, position 4, position 23, position 28, or position 29 of the sense strand, and wherein the position is from 5 'To 3' number. In some aspects, the tissue is the hypothalamus, wherein the at least one lipid moiety is associated with position 1, position 4, position 12, position 13, position 18, position 20, position 23, position 28, position 29 of the sciatic strand , or the nucleotide at position 30 binds, and wherein the positions are numbered from 5' to 3'. In other aspects, the tissue is frontal cortex, wherein the at least one lipid moiety binds to nucleotide at position 23 of the sense strand, and wherein the positions are numbered from 5' to 3'. In any of the foregoing or related aspects of the methods described herein, a single dose of the oligonucleotide or pharmaceutical composition reduces the expression of stellate cell mRNA for at least 4 weeks, at least 8 weeks, at least 12 weeks, at least 23 weeks weeks, at least 26 weeks, or at least 29 weeks. In some aspects, a single dose of an oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for at least 4 weeks. In some aspects, a single dose of an oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for at least 8 weeks. In some aspects, a single dose of an oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for at least 12 weeks. In some aspects, a single dose of an oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for at least 23 weeks. In some aspects, a single dose of an oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for at least 26 weeks. In some aspects, a single dose of an oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for at least 29 weeks. In any of the foregoing or related aspects of the methods described herein, a single dose of the oligonucleotide or pharmaceutical composition reduces the expression of stellate cell mRNA by up to 4 weeks, by 8 weeks, by 12 weeks , up to 23 weeks, up to 26 weeks, or up to 29 weeks. In some aspects, a single dose of oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for up to 4 weeks. In some aspects, a single dose of oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for up to 8 weeks. In some aspects, a single dose of oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for up to 12 weeks. In some aspects, a single dose of oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for up to 23 weeks. In some aspects, a single dose of oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for up to 26 weeks. In some aspects, a single dose of oligonucleotide or pharmaceutical composition reduces stellate cell mRNA expression for up to 29 weeks. In any of the foregoing or related aspects of the methods described herein, a single dose of an oligonucleotide or pharmaceutical composition reduces the expression of stellate cell mRNA for up to one year. In some aspects, the present disclosure provides kits comprising an oligonucleotide described herein, optionally a pharmaceutically acceptable carrier, and a protein for administration to a patient with stellate cell mRNA. Drug instructions for individuals exhibiting relevant diseases, illnesses, or conditions. In some forms, the drug package insert includes instructions for intrathecal administration. In other aspects, the present disclosure provides use of an oligonucleotide or pharmaceutical composition described herein for the manufacture of a medicament for the treatment of a disease, disorder, or condition associated with the expression of stellate cell mRNA. In a further aspect, the present disclosure provides oligonucleotides or pharmaceutical compositions described herein for use in, or may be suitable for use in, a medicament for the treatment of diseases, disorders, or conditions associated with the expression of stellate cell mRNA purpose.

In some aspects, the present disclosure provides oligonucleotide-lipid conjugates (e.g., RNAi oligonucleotide-lipid conjugates) that reduce expression of a target gene expressed in stellate cells in the central nervous system (CNS) . In other aspects, the present disclosure provides methods of treating diseases or conditions associated with expression of stellate cell mRNA (eg, diseases of the CNS). In other aspects, the present disclosure provides use of lipid-conjugated RNAi oligonucleotides described herein, or pharmaceutically acceptable compositions thereof, to treat diseases or conditions associated with expression of stellate cell mRNA (e.g., neurological disease and/or inappropriate genetic expression). In other aspects, the present disclosure provides methods of using lipid-conjugated RNAi oligonucleotides described herein in the manufacture of medicaments for treating diseases or conditions associated with expression of stellate cell mRNA. In other aspects, lipid-conjugated RNAi oligonucleotides provided herein are used to modulate (e.g., inhibit or reduce) stellate cell target genes associated with neurological diseases or disorders in the CNS. Manifested to treat neurological diseases or conditions. In some aspects, the present disclosure provides methods of treating neurological diseases or disorders by reducing expression of stellate cell target genes associated with the neurological disease or disorder in the CNS (e.g., in cells, tissues, or organs of the CNS) . Lipids - Combined RNAi OligonucleotidesIn particular, the present disclosure provides lipid-conjugated RNAi oligonucleotides (eg, RNAi oligonucleotide-lipid conjugates) that reduce expression of stellate cell target genes in the CNS. In some embodiments, lipid-conjugated RNAi oligonucleotides provided by the present disclosure are targeted to mRNA encoding a target gene. The messenger RNA (mRNA) encoding the target gene and targeted by the lipid-conjugated RNAi oligonucleotides of the present disclosure is referred to herein as "target mRNA". mRNA target sequence In some embodiments, lipid-conjugated RNAi oligonucleotides target target sequences comprising target stellate cell mRNA. In some embodiments, lipid-conjugated RNAi oligonucleotides target target sequences within target stellate cell mRNA. In some embodiments, a lipid-conjugated RNAi oligonucleotide, or a portion, fragment, or strand thereof (e.g., an antisense or guide strand of a double-stranded oligonucleotide) is combined with a stellate cell mRNA of interest. Target sequences are spliced or glued, thereby reducing target gene expression. In some embodiments, lipid-conjugated RNAi oligonucleotides target a target sequence comprising target stellate cell mRNA for the purpose of reducing expression of the stellate cell target gene in vivo. In some embodiments, the amount or degree of reduction in target gene expression by a lipid-conjugated RNAi oligonucleotide that targets a specific stellate cell target sequence correlates with the efficacy of the lipid-conjugated RNAi oligonucleotide. Union. In some embodiments, reducing the amount or extent of target gene expression by lipid-conjugated RNAi oligonucleotides targeting specific stellate cell target sequences is consistent with treatment with lipid-conjugated RNAi oligonucleotides. Correlates the amount or degree of therapeutic benefit to an individual or patient suffering from a disease, disorder, or condition associated with expression of a target gene. Certain nucleosides have been discovered by examining the nucleotide sequences of mRNAs encoding target genes, including those from a variety of different species (e.g., humans, Malay monkeys, mice, and rats) and the results of in vitro and in vivo tests Acid sequences and certain systematic modifications to those oligonucleotides are more suitable than other oligonucleotide sequences for the reduction of RNAi oligonucleotide mediators, and therefore they can be used as additional oligos targeting specific gene target sequences. part of a nucleotide. In some embodiments, the sense strand of a lipid-conjugated RNAi oligonucleotide described herein, or a portion or fragment thereof, comprises a target sequence similar to that comprising a stellate cell target mRNA (e.g., having no more than 4 mismatch) or identical nucleotide sequences. In some embodiments, a portion or region of the sense strand of a double-stranded oligonucleotide described herein includes a target sequence including a stellate cell target mRNA. In some embodiments, the stellate cell mRNA target sequence is associated with acute or chronic pain. In some embodiments, stellate cell mRNA target sequences are associated with neurological disorders. In some embodiments, the stellate cell mRNA target sequence is an mRNA expressed in stellate cells in at least one region of the CNS. In some embodiments, the stellate cell mRNA target sequence is an mRNA expressed in stellate cells of the spinal cord. In some embodiments, the stellate cell mRNA target sequence is an mRNA expressed in stellate cells of the lumbar spinal cord. In some embodiments, the stellate cell mRNA target sequence is an mRNA expressed in stellate cells of the thoracic spinal cord. In some embodiments, the stellate cell mRNA target sequence is an mRNA expressed in stellate cells of the cervical spinal cord. In some embodiments, the stellate cell mRNA target sequence is an mRNA expressed in stellate cells of the hypothalamus. In some embodiments, the stellate cell mRNA target sequence is an mRNA expressed in stellate cells in the medulla oblongata. In some embodiments, the stellate cell mRNA target sequence is an mRNA expressed in stellate cells of the hippocampus. In some embodiments, the stellate cell mRNA target sequence is an mRNA expressed in cerebellar stellate cells. In some embodiments, the stellate cell mRNA target sequence is an mRNA expressed in stellate cells of the frontal cortex. In some embodiments, the stellate cell mRNA target sequence is an mRNA associated with a disease, disorder, or condition of the CNS. In some embodiments, the oligonucleotides herein have GFAPRegions of mRNA complementarity (e.g., stellate cell mRNA target sequences are GFAPsequence. In some embodiments, the target sequence is tethered to GFAPwithin the target sequence of the mRNA) to achieve the purpose of targeting the mRNA in cells and inhibiting its expression. In some embodiments, the oligonucleotides herein comprise GFAPTargeting sequences (e.g., antisense or leader strands of double-stranded oligonucleotides) that have the ability to bind to GFAPThe complementary region to which a target sequence binds or adheres. In some embodiments, a portion or region of the sense strand of a double-stranded oligonucleotide described herein comprises GFAPtarget sequence. In some specific embodiments, GFAPThe target sequence includes or consists of the nucleotide sequence of any one of SEQ ID NOs: 28 to 31. RNAi Oligonucleotide targeting sequence In some embodiments, lipid-bound RNAi oligonucleotides provided by the present disclosure comprise targeting sequences. As used herein, the term "targeting sequence" refers to a nucleotide sequence having a region complementary to a specific nucleotide sequence comprising an mRNA (eg, a stellate cell target mRNA). In some embodiments, lipid-binding RNAi oligonucleotides provided by the present disclosure comprise a gene-targeting sequence having a region complementary to a nucleotide sequence comprising a target sequence of target mRNA. In some embodiments, the targeting sequence is a stellate cell mRNA target sequence. The targeting sequence confers the ability of the lipid-conjugated RNAi oligonucleotide to specifically target the mRNA by joining or adhering to the target sequence containing the target mRNA via complementary (Wasen-Click) base pairing. In some embodiments, lipid-conjugated RNAi oligonucleotides (or strands thereof, e.g., antisense strands or guide strands of double-stranded oligonucleotides) herein comprise compounds that possess the properties of (Rick) base-pairing to a complementary region of the targeting sequence that joins or binds to the target sequence containing the stellate cell target mRNA. In some embodiments, lipid-conjugated RNAi oligonucleotides (or strands thereof, e.g., antisense strands or guide strands of double-stranded oligonucleotides) herein comprise compounds that possess the properties of (Rick) target sequence of the complementary region that base-pairs with or binds to the target sequence in the stellate cell target mRNA. The targeting sequence is typically of appropriate length and base content to enable the lipid-conjugated RNAi oligonucleotide (or strand thereof) to engage or bind to the specific target mRNA (e.g., stellate cell mRNA) to achieve the inhibitory target The purpose of gene expression. In some embodiments, the targeting sequence is at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21 , at least about 22, at least about 23, at least about 24, at least about 25, at least about 26, at least about 27, at least about 28, at least about 29, or at least about 30 nucleotides long. In some embodiments, the targeting sequence is at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 nucleotides. In some embodiments, the targeting sequences are about 12 to about 30 (eg, 12 to 30, 12 to 22, 15 to 25, 17 to 21, 18 to 27, 19 to 27, or 15 to 30) Nucleotide length. In some embodiments, the targeting sequence is about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides long. In some embodiments, the targeting sequence is 18 nucleotides long. In some embodiments, the targeting sequence is 19 nucleotides long. In some embodiments, the targeting sequence is 20 nucleotides long. In some embodiments, the targeting sequence is 21 nucleotides long. In some embodiments, the targeting sequence is 22 nucleotides long. In some embodiments, the targeting sequence is 23 nucleotides long. In some embodiments, the targeting sequence is 24 nucleotides long. In some embodiments, lipid-conjugated RNAi oligonucleotides herein comprise a targeting sequence that is fully complementary to a target sequence comprising a stellate cell target mRNA. In some embodiments, lipid-conjugated RNAi oligonucleotides herein comprise a targeting sequence that is fully complementary to a target sequence within a stellate cell target mRNA. In some embodiments, the targeting sequence is partially complementary to the target sequence comprising the target mRNA. In some embodiments, the targeting sequence is partially complementary to the target sequence within the stellate cell target mRNA. In some embodiments, the targeting sequence includes a region of contiguous nucleotides that includes the antisense strand. In some embodiments, lipid-conjugated RNAi oligonucleotides herein comprise a targeting sequence complementary to a contiguous sequence of nucleotides comprising a stellate cell target mRNA, wherein the contiguous sequence of nucleotides is about 12 to about 30 nucleotides long (e.g., 12 to 30, 12 to 28, 12 to 26, 12 to 24, 12 to 20, 12 to 18, 12 to 16, 14 to 22, 16 to 20, 18 to 20 , or 18 to 19 nucleotides long). In some embodiments, the lipid-bound RNAi oligonucleotide comprises a targeting sequence complementary to a contiguous sequence of nucleotides comprising a stellate cell target mRNA, wherein the contiguous sequence of nucleotides is 10, 11, 12 , 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides long. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a targeting sequence that is complementary to a contiguous sequence of nucleotides comprising the target mRNA, wherein the contiguous sequence of nucleotides is 15 nucleotides long. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a targeting sequence that is complementary to a contiguous sequence of nucleotides comprising the target mRNA, wherein the contiguous sequence of nucleotides is 19 nucleotides long. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a targeting sequence complementary to a contiguous sequence of nucleotides comprising a stellate cell target mRNA, wherein the contiguous sequence of nucleotides is 15 nucleotides long. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a targeting sequence complementary to a contiguous sequence of nucleotides comprising a stellate cell target mRNA, wherein the contiguous sequence of nucleotides is 19 nucleotides long. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is completely complementary (e.g., without mismatches) to the target sequence comprising the stellate cell target mRNA and includes the full length of the antisense strand. . In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is completely complementary (e.g., without mismatches) to the target sequence comprising the stellate cell target mRNA and includes the full length of the antisense strand. part of it. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is completely complementary (e.g., no mismatch) to the target sequence comprising the stellate cell target mRNA and includes 10 of the antisense strands. to 20 nucleotides. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is completely complementary (e.g., no mismatch) to the target sequence comprising the stellate cell target mRNA and includes the antisense strand. to 19 nucleotides. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is completely complementary (e.g., no mismatch) to the target sequence comprising the stellate cell target mRNA and includes the antisense strand. nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, or 22 nucleotides. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is completely complementary (e.g., no mismatch) to the target sequence comprising the stellate cell target mRNA and includes the antisense strand. nucleotides. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is completely complementary (e.g., no mismatch) to the target sequence comprising the stellate cell target mRNA and includes 20 of the antisense strand. nucleotides. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is partially complementary (e.g., with no more than 4 mismatches) to a target sequence comprising a stellate cell target mRNA and includes an anti- The total length of the stock. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is partially complementary (e.g., with no more than 4 mismatches) to a target sequence comprising a stellate cell target mRNA and includes an anti- Part of the total length of the stock. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is partially complementary (e.g., with no more than 4 mismatches) to a target sequence comprising a stellate cell target mRNA and includes an anti- 10 to 20 nucleotides of the strand. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is partially complementary (e.g., with no more than 4 mismatches) to a target sequence comprising a stellate cell target mRNA and includes an anti- 15 to 19 nucleotides of the strand. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is partially complementary (e.g., with no more than 4 mismatches) to a target sequence comprising a stellate cell target mRNA and includes an anti- 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, or 22 nucleosides acid. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is partially complementary (e.g., with no more than 4 mismatches) to a target sequence comprising a stellate cell target mRNA and includes an anti- The 19 nucleotides of the righteous stock. In some embodiments, the targeting sequence of the lipid-binding RNAi oligonucleotides herein is partially complementary (e.g., with no more than 4 mismatches) to a target sequence comprising a stellate cell target mRNA and includes an anti- The 20 nucleotides of the righteous stock. In some specific embodiments, lipid-binding RNAi oligonucleotides herein comprise a targeting sequence that has one or more base pairing (bp) mismatches with a corresponding target sequence comprising a stellate cell target mRNA. In some embodiments, the targeting sequence has a 1 bp mismatch, a 2 bp mismatch, a 3 bp mismatch, a 4 bp mismatch, or a 5 bp mismatch with a corresponding target sequence comprising a stellate cell target mRNA. Mismatch, provided that the ability of the targeting sequence to engage or bind to the target sequence and/or the ability of the lipid-bound RNAi oligonucleotide to inhibit or reduce expression of the target gene is maintained under appropriate hybridization conditions (e.g., under physiological conditions ). Alternatively, in some embodiments, the targeting sequence and the corresponding target sequence comprising the stellate cell target mRNA comprise no more than 1, no more than 2, no more than 3, no more than 4, or no more than 5 bp mismatch, provided that the ability of the targeting sequence to engage or adhere to the target sequence and/or the ability of the lipid-bound RNAi oligonucleotide to inhibit or reduce the expression of the target gene is maintained under appropriate hybridization conditions. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a targeting sequence that has 1 mismatch to the corresponding target sequence. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a targeting sequence that has 2 mismatches with the corresponding target sequence. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a targeting sequence that has 3 mismatches to the corresponding target sequence. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a targeting sequence that has 4 mismatches to the corresponding target sequence. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a targeting sequence that has 5 mismatches to the corresponding target sequence. In some embodiments, the lipid-binding RNAi oligonucleotide comprises a targeting sequence that has more than one mismatch (e.g., 2, 3, 4, 5 or more mismatches) with a corresponding target sequence, wherein the At least 2 (e.g., all) of the mismatches are located contiguously (e.g., 2, 3, 4, 5 or more mismatches in a row), or wherein the mismatches are interspersed throughout the targeted sequence. in any position. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a targeting sequence that has more than one mismatch (e.g., 2, 3, 4, 5 or more mismatches) with a corresponding target sequence, wherein At least 2 (e.g., all) of the mismatches are located contiguously (e.g., 2, 3, 4, 5 or more mismatches in succession), or at least one or more of the non-mismatched base pairs The system is located between mismatches, or a combination thereof. Type of oligonucleotide In the methods herein, a variety of RNAi oligonucleotide types and/or structures can be used to reduce target gene expression (eg, reduce expression of a target gene expressed in stellate cells). Any RNAi oligonucleotide type described herein or elsewhere is contemplated for use as a framework for incorporation into targeting sequences herein for the purpose of inhibiting or reducing expression of the corresponding target gene in stellate cells in the CNS. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein inhibit target gene expression by participating in the upstream or downstream RNA interference (RNAi) pathway of Dicer. For example, RNAi oligonucleotides have been developed in which each strand is approximately 19 to 25 nucleotides in size and has at least one 3' overhang of 1 to 5 nucleotides (see, eg, U.S. Patent No. 8,372,968). Longer oligonucleotides that are processed by Dicer to generate active RNAi products have also been developed (see, eg, US Pat. No. 8,883,996). Further work resulted in extended double-stranded oligonucleotides in which at least one end of at least one strand extends beyond the double-stranded targeting region, including wherein one of the strands includes a thermodynamically stable The structure of the four-ring structure (see, for example, U.S. Patent Nos. 8,513,207 and 8,927,705, and International Patent Application Publication No. WO 2010/033225). Such structures may include single-stranded extensions (on one or both sides of the molecule) as well as double-stranded extensions. In some embodiments, the RNAi oligonucleotide conjugates herein participate in the RNAi pathway downstream of Dicer engagement (eg, Dicer cleavage). In some embodiments, the oligonucleotides described herein are Dicer substrates. In some embodiments, after endogenous Dicer processing, a 19 to 23 nucleotide long double-stranded nucleic acid is produced that is capable of reducing expression of the stellate cell target mRNA. In some embodiments, the lipid-bound RNAi oligonucleotide has an overhang in the 3' end of the sense strand (eg, is 1, 2, or 3 nucleotides long). In some embodiments, lipid-conjugated RNAi oligonucleotides (e.g., siRNA conjugates) comprise a 21 nucleotide leader strand antisense to a stellate cell target mRNA and a complementary follower strand, where the two strands are bonded to A 19-bp duplex is formed with 2 nucleotide overhangs at either or both 3' ends. Longer oligonucleotide designs may also be considered, including those with a 23-nt leader strand and a 21-nt follower strand, where the oligonucleotide on the right side of the molecule (the 3' end of the follower strand/ There is a blunt end on the 5' end of the leader strand) and a two-nucleotide 3' leader overhang on the left side of the molecule (5' end of the follower strand/3' end of the leader strand). In this type of molecule, a 21 bp double-stranded region is present. See, for example, U.S. Patent Nos. 9,012,138; 9,012,621; and 9,193,753. In some embodiments, the RNAi oligonucleotide conjugates disclosed herein comprise each are about 17 to 26 (e.g., 17 to 26, 20 to 25, or 21 to 23) nucleotides in length. Just shares and anti-righteous shares within the scope. In some embodiments, lipid-conjugated RNAi oligonucleotides disclosed herein comprise a sense strand and an antisense strand each ranging from about 19 to 22 nucleotides in length. In some embodiments, the sense strand and antisense strand are of equal length. In some embodiments, lipid-conjugated RNAi oligonucleotides disclosed herein comprise a sense strand and an antisense strand, such that there is 3 on the sense strand or the antisense strand, or both the sense strand and the antisense strand. 'Protruding end. In some embodiments, for lipid-bound RNAi oligonucleotides having a sense strand and an antisense strand both in the range of about 21 to 23 nucleotides in length, the sense strand, the antisense strand , or the 3' overhangs on both the sense and antisense strands are 1 or 2 nucleotides long. In some embodiments, a lipid-conjugated RNAi oligonucleotide has a 22 nucleotide leader strand and a 20 nucleotide follower strand, where on the right side of the molecule (the 3' end of the follower/leader There is a blunt end on the 5' end of the molecule, and a 2-nucleotide 3'-leader overhang on the left side of the molecule (5' end of the follower strand/3' end of the leader strand). In this type of molecule, a 20 bp double-stranded region is present. Other RNAi oligonucleotide designs for use with the compositions and methods herein include: 16-mer siRNA (see, e.g., Nucleic Acids in Chemistry and Biology, Blackburn(ed.), ROYAL SOCIETY OF CHEMISTRY, 2006), shRNA (e.g., with a backbone of 19 bp or less; see e.g., Moore et al.(2010) METHODS MOL. BIOL. 629:141-58), blunt siRNA (e.g., 19 bp long; see e.g. Kraynack & Baker (2006) RNA 12:163-76), asymmetric siRNA (aiRNA; see e.g., Sun et al.(2008) NAT. BIOTECHNOL .26: 1379-82), asymmetric shorter duplex siRNA (see e.g., Chang et al .(2009) MOL. Ther. 17:725-32), forked siRNA (see e.g., Hohjoh (2004) FEBS Lett. 557:193-98), and small internal segmented interfering RNA (siRNA; see e.g., Bramsen et al.(2007) NUCLEIC ACIDS RES. 35:5886-97). Further non-limiting examples of oligonucleotide structures that may be used in some embodiments to reduce or inhibit the expression of a gene of interest are microRNAs (miRNAs), short hairpin RNAs (shRNAs), and short siRNAs (see e.g. ,Hamilton et al.(2002) EMBO J. 21:4671-79; see also, U.S. Patent Application Publication No. 2009/0099115). Antisense stocks In some embodiments, the antisense strand of the lipid-conjugated RNAi oligonucleotide is referred to as the "lead strand." For example, the antisense strand engages the RNA-induced silencing complex (RISC) and interacts with Argu ( Argonaute) protein such as Ago2 engages, or engages or engages with one or more similar factors, and directs the silencing of the target gene, so the antisense strand is called a guide strand. In some embodiments, the justice strands that are complementary to the leading strands are called "follower strands." In some specific embodiments, lipid-conjugated RNAi oligonucleotides herein comprise up to about 50 nucleotides in length (e.g., up to 50, up to 40, up to 35, up to 30, up to 27, up to 25, up to 21. Antisense strands at most 19, at most 17, at most 15, or at most 12 nucleotides long). In some embodiments, the lipid-bound RNAi oligonucleotide comprises at least about 12 nucleotides long (e.g., at least 12, at least 15, at least 19, at least 21, at least 22, at least 25, at least 27, at least 30. Antisense strands at least 35, or at least 38 nucleotides long). In some embodiments, between about 12 to about 40 (e.g., 12 to 40, 12 to 36, 12 to 32, 12 to 28, 15 to 40, 15 to 36, 15 to 32, 15 to Antisense in the range of 30, 15 to 28, 17 to 22, 17 to 25, 19 to 27, 19 to 30, 20 to 40, 22 to 40, 25 to 40, or 32 to 40) nucleotides long share. In some embodiments, lipid-conjugated RNAi oligonucleotides herein comprise antisense strands that are 15 to 30 nucleotides long. In some specific embodiments, the antisense strands of any of the lipid-conjugated RNAi oligonucleotides disclosed herein have 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 nucleotides long. In some embodiments, lipid-conjugated RNAi oligonucleotides comprise antisense strands that are 19 to 23 nucleotides long. In some embodiments, the lipid-conjugated RNAi oligonucleotide comprises a 19 nucleotide long antisense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotide comprises a 20 nucleotide long antisense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotide comprises a 21 nucleotide long antisense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotide comprises a 22 nucleotide long antisense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotide comprises a 23 nucleotide long antisense strand. justice unit In some specific embodiments, lipid-conjugated RNAi oligonucleotides disclosed herein comprise up to about 50 nucleotides long (e.g., up to 50, up to 40, up to 36, up to 30, up to 27, up to 25 , up to 21, up to 19, up to 17, or up to 12 nucleotides long) sense strand (or follower strand). In some specific embodiments, lipid-conjugated RNAi oligonucleotides herein comprise at least about 12 nucleotides in length (e.g., at least 12, at least 15, at least 19, at least 21, at least 25, at least 27, at least 30. A sense strand at least 36, or at least 38 nucleotides long). In some embodiments, the lipid-binding RNAi oligonucleotides herein comprise from about 12 to about 50 (e.g., 12 to 50, 12 to 40, 12 to 36, 12 to 32, 12 to 28, 15 to 40, 15 to 36, 15 to 32, 15 to 28, 17 to 21, 17 to 25, 19 to 27, 19 to 30, 20 to 40, 22 to 40, 25 to 40, or 32 to 40) nucleotide length range of the sense strand. In some embodiments, lipid-conjugated RNAi oligonucleotides herein comprise a sense strand that is 15 to 50 nucleotides long. In some embodiments, lipid-conjugated RNAi oligonucleotides herein comprise a sense strand that is 18 to 36 nucleotides long. In some embodiments, lipid-conjugated RNAi oligonucleotides herein include 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 cores The long righteous stock of glycosides. In some embodiments, lipid-conjugated RNAi oligonucleotides herein comprise a sense strand that is 17 to 21 nucleotides long. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 17 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a sense strand that is 18 nucleotides long. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 19 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 20 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 21 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 22 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 23 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 24 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 25 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 26 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 27 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 28 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 29 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 30 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 31 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 32 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 33 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 34 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 35 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a 36 nucleotide long sense strand. In some embodiments, the sense strand contains a backbone-loop structure at its 3' end. In some embodiments, the backbone-loops are formed by intrastrand base pairing. In some embodiments, the sense strand contains a backbone-loop structure at its 5' end. In some embodiments, the backbone is a duplex that is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 nucleotides long. In some embodiments, the backbone of the backbone-loop includes a duplex that is 2 nucleotides long. In some embodiments, the backbone of the backbone-loop includes a 3 nucleotide long duplex. In some embodiments, the backbone of the backbone-loop includes a duplex that is 4 nucleotides long. In some embodiments, the backbone of the backbone-loop includes a 5 nucleotide long duplex. In some embodiments, the backbone of the backbone-loop includes a duplex that is 6 nucleotides long. In some embodiments, the backbone of the backbone-loop includes a 7 nucleotide long duplex. In some embodiments, the backbone of the backbone-loop includes an 8 nucleotide long duplex. In some embodiments, the backbone of the backbone-loop includes a 9 nucleotide long duplex. In some embodiments, the backbone of the backbone-loop includes a duplex that is 10 nucleotides long. In some embodiments, the backbone of the backbone-loop includes a duplex that is 11 nucleotides long. In some embodiments, the backbone of the backbone-loop includes a duplex that is 12 nucleotides long. In some embodiments, the stem-loop stem includes a duplex that is 13 nucleotides long. In some embodiments, the backbone of the backbone-loop includes a duplex that is 14 nucleotides long. In some embodiments, the backbone-loop provides lipid-bound RNAi oligonucleotides with protection against degradation (e.g., enzymatic degradation), promotes or improves targeting and/or delivery to target cells, tissues, or organs, Or both. For example, in some embodiments, loops of backbone-loops provide nucleotides that include one or more modifications that promote, improve, or increase response to a target mRNA (e.g., a target mRNA expressed in the CNS) Targeting, inhibition of target gene expression, and/or delivery to target cells, tissues, or organs (e.g., CNS), or combinations thereof. In some embodiments, the backbone-loop itself or the modification(s) to the backbone loop does not substantially affect the inherent gene expression suppressing activity of the lipid-bound RNAi oligonucleotide, but promotes, improves, or increases Stability (eg, providing protection against degradation) and/or delivery of lipid-bound RNAi oligonucleotides to target cells, tissues, or organs (eg, CNS). In certain embodiments, lipid-conjugated RNAi oligonucleotides herein comprise a sense strand comprising (e.g., at its 3' end) a backbone-loop as shown: S1-L-S2 , wherein S1 is complementary to S2, and wherein L is up to about 10 nucleotides long (e.g., 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides long) between S1 and S2 ) of single-stranded loops. In some embodiments, loop (L) is 3 nucleotides long. In some embodiments, loop (L) is 4 nucleotides long. In some specific embodiments, the tetraloop contains the sequence 5'-GAAA-3'. In some embodiments, the backbone loop includes the sequence 5'-GCAGCCGAAA GGCUGC-3' (SEQ ID NO: 32). In some embodiments, the loop (L) having the backbone-loop structure S1-L-S2 as described above is a tricyclic ring. In some embodiments, tricycles include ribonucleotides, deoxyribonucleotides, modified nucleotides, delivery ligands, and combinations thereof. In some embodiments, the loop (L) of the backbone-loop having the structure S1-L-S2 as described above is a four-ring (eg, within a notched four-ring structure). In some embodiments, the tetracycle includes ribonucleotides, deoxyribonucleotides, modified nucleotides, delivery ligands, and combinations thereof. In some embodiments, the loop (L) of the backbone-loop having the structure S1-L-S2 as described above is a tetracyclic loop as described in U.S. Patent No. 10,131,912, which is incorporated by reference ( For example, within a gapped four-ring structure). Duplex length In some embodiments, there are at least 12 double-stranded systems formed between the sense strand and the antisense strand (eg, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, or at least 21) Nucleotide length. In some embodiments, the duplex system formed between the sense strand and the antisense strand ranges from 12 to 30 nucleotides in length (e.g., 12 to 30, 12 to 27, 12 to 22, 15 to 25, 18 to 30, 18 to 22, 18 to 25, 18 to 27, 18 to 30, 19 to 30, or 21 to 30 nucleotides long). In some specific embodiments, a double-stranded system 12, 13, 14, 15, 16, 17, 18, 19, 29, 21, 22, 23, 24, 25, 26 is formed between the sense strand and the antisense strand. , 27, 28, 29 or 30 nucleotides long. In some embodiments, the duplex formed between the sense strand and the antisense strand is 15 to 30 base pairs long. In some embodiments, the duplex formed between the sense strand and the antisense strand is 17 to 21 base pairs long. In some embodiments, the duplex formed between the sense strand and the antisense strand is 17 base pairs long. In some embodiments, the duplex formed between the sense strand and the antisense strand is 18 base pairs long. In some embodiments, the duplex formed between the sense strand and the antisense strand is 19 base pairs long. In some embodiments, the duplex formed between the sense strand and the antisense strand is 20 base pairs long. In some embodiments, the duplex formed between the sense strand and the antisense strand is 21 base pairs long. In some embodiments, the duplex formed between the sense strand and the antisense strand does not span the entire length of the sense strand and/or the antisense strand. In some embodiments, the duplex between the sense strand and the antisense strand spans the entire length of the sense strand or antisense strand. In some embodiments, the duplex between the sense strand and the antisense strand spans the entire length of both the sense strand and the antisense strand. oligonucleotide end In some embodiments, lipid-conjugated RNAi oligonucleotides disclosed herein comprise a sense strand and an antisense strand, such that there is 3 on the sense strand or the antisense strand, or both the sense strand and the antisense strand. 'Protruding end. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein have a 5' end that is thermodynamically less stable than the other 5' end. In some embodiments, asymmetric lipid-bound RNAi oligonucleotides are provided that include a blunt end at the 3' end of the sense strand and an overhang at the 3' end of the antisense strand. In some embodiments, the 3' overhang on the antisense strand is 1 to 4 nucleotides long (eg, 1, 2, 3, or 4 nucleotides long). In some specific embodiments, the 3'-overhang is about one (1) to twenty (20) nucleotides long (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9 , 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or about 20 nucleotides long). In some embodiments, the 3' overhangs range from about one (1) to nineteen (19), from one (1) to eighteen (18), from one (1) to seventeen (17), (1) to sixteen (16), one (1) to fifteen (15), one (1) to fourteen (14), one (1) to thirteen (13), one (1) ) to twelve (12), one (1) to eleven (11), one (1) to ten (10), one (1) to nine (9), one (1) to eight (8), one (1) to seven (7), one (1) to six (6), one (1) to five (5), one (1) to four (4), one ( 1) to three (3), or about one (1) to two (2) nucleotides in length. In some embodiments, the 3'-overhang is (1) nucleotide long. In some embodiments, the 3'-overhang is two (2) nucleotides long. In some embodiments, the 3'-overhang is three (3) nucleotides long. In some embodiments, the 3'-overhang is four (4) nucleotides long. In some embodiments, the 3'-overhang is five (5) nucleotides long. In some embodiments, the 3'-overhang is six (6) nucleotides long. In some embodiments, the 3'-overhang is seven (7) nucleotides long. In some embodiments, the 3'-overhang is eight (8) nucleotides long. In some embodiments, the 3'-overhang is nine (9) nucleotides long. In some embodiments, the 3'-overhang is ten (10) nucleotides long. In some embodiments, the 3'-overhang is eleven (11) nucleotides long. In some embodiments, the 3'-overhang is twelve (12) nucleotides long. In some embodiments, the 3'-overhang is thirteen (13) nucleotides long. In some embodiments, the 3'-overhang is fourteen (14) nucleotides long. In some embodiments, the 3'-overhang is fifteen (15) nucleotides long. In some embodiments, the 3'-overhang is sixteen (16) nucleotides long. In some embodiments, the 3'-overhang is seventeen (17) nucleotides long. In some embodiments, the 3'-overhang is eighteen (18) nucleotides long. In some embodiments, the 3'-overhang is nineteen (19) nucleotides long. In some embodiments, the 3'-overhang is twenty (20) nucleotides long. Generally, oligonucleotides used for RNAi have a two (2) nucleotide overhang on the 3' end of the antisense (leader) strand. However, other protrusions are also possible. In some embodiments, the overhang is a 3' overhang, which is comprised between one and four nucleotides in length, optionally one to four, one to three, one to two, two to four, two to three, or one, two, three, or four nucleotides. In some embodiments, the overhang is a 5' overhang, which is comprised between one and four nucleotides in length, optionally one to four, one to three, one to two, two to four, two to three, or one, two, three, or four nucleotides. In some embodiments, the oligonucleotides herein comprise a sense strand and an antisense strand, wherein the 5' end of either or both strands includes a 5'-overhang comprising one or more nucleotides. In some embodiments, the oligonucleotides herein comprise a sense strand and an antisense strand, wherein the sense strand comprises a 5'-overhang comprising one or more nucleotides. In some embodiments, the oligonucleotides herein comprise a sense strand and an antisense strand, wherein the antisense strand comprises a 5'-overhang comprising one or more nucleotides. In some embodiments, oligonucleotides herein comprise a sense strand and an antisense strand, wherein both the sense strand and the antisense strand comprise a 5'-overhang comprising one or more nucleotides. In some embodiments, the 5'-overhang is about one (1) to twenty (20) nucleotides long (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9 , 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or about 20 nucleotides long). In some embodiments, the 5' overhangs range from about one (1) to nineteen (19), from one (1) to eighteen (18), from one (1) to seventeen (17), (1) to sixteen (16), one (1) to fifteen (15), one (1) to fourteen (14), one (1) to thirteen (13), one (1) ) to twelve (12), one (1) to eleven (11), one (1) to ten (10), one (1) to nine (9), one (1) to eight (8), one (1) to seven (7), one (1) to six (6), one (1) to five (5), one (1) to four (4), one ( 1) to three (3), or about one (1) to two (2) nucleotides in length. In some embodiments, the 5'-overhang is (1) nucleotide long. In some embodiments, the 5'-overhang is two (2) nucleotides long. In some embodiments, the 5'-overhang is three (3) nucleotides long. In some embodiments, the 5'-overhang is four (4) nucleotides long. In some embodiments, the 5'-overhang is five (5) nucleotides long. In some embodiments, the 5'-overhang is six (6) nucleotides long. In some embodiments, the 5'-overhang is seven (7) nucleotides long. In some embodiments, the 5'-overhang is eight (8) nucleotides long. In some embodiments, the 5'-overhang is nine (9) nucleotides long. In some embodiments, the 5'-overhang is ten (10) nucleotides long. In some embodiments, the 5'-overhang is eleven (11) nucleotides long. In some embodiments, the 5'-overhang is twelve (12) nucleotides long. In some embodiments, the 5'-overhang is thirteen (13) nucleotides long. In some embodiments, the 5'-overhang is fourteen (14) nucleotides long. In some embodiments, the 5'-overhang is fifteen (15) nucleotides long. In some embodiments, the 5'-overhang is sixteen (16) nucleotides long. In some embodiments, the 5'-overhang is seventeen (17) nucleotides long. In some embodiments, the 5'-overhang is eighteen (18) nucleotides long. In some embodiments, the 5'-overhang is nineteen (19) nucleotides long. In some embodiments, the 5'-overhang is twenty (20) nucleotides long. In some embodiments, one or more (eg, 2, 3, or 4) terminal nucleotides at the 3' end or the 5' end of the sense strand and/or the antisense strand are modified. For example, in some embodiments, one or both terminal nucleotides at the 3' end of the antisense strand are modified. In some embodiments, the last nucleotide at the 3' end of the antisense strand is modified, e.g., includes a 2' modification, e.g., 2'-O-methoxyethyl. In some embodiments, the last one or two terminal nucleotides at the 3' end of the antisense strand are complementary to the target. In some embodiments, the last one or two nucleotides at the 3' end of the antisense strand are not complementary to the target. In some embodiments, the RNAi oligonucleotide conjugates disclosed herein comprise a backbone-loop structure at the 3' end of the sense strand and two terminal overhang cores at the 3' end of the antisense strand. glycosides. In some embodiments, the RNAi oligonucleotide conjugates herein comprise a gapped tetracyclic structure, wherein the 3' end of the sense strand contains a backbone tetracyclic structure and the 3' end of the antisense strand contains two Terminal overhang nucleotide. In some embodiments, the overhang is selected from AA, GG, AG, and GA. In some embodiments, the overhang is AA. In some embodiments, the overhang is AG. In some embodiments, the overhang is GA. In some embodiments, the two terminal overhang nucleotides are GG. Generally, one or both of the two terminal GG nucleotides of the antisense strand are not complementary to the target. In some embodiments, the 5' end and/or 3' end of the sense strand or antisense strand has an inverted cap nucleotide. In some embodiments, one or more (e.g., 2, 3, 4, 5, 6) are provided between the terminal nucleotides at the 3' end or 5' end of the sense strand and/or the antisense strand. Modified inter-nucleotide linkages. In some embodiments, modified internucleotide linkages are provided between overhangs at the 3' or 5' ends of the sense strand and/or antisense strand. Oligonucleotide modification In some embodiments, RNAi oligonucleotide conjugates disclosed herein comprise one or more modifications. Oligonucleotides (e.g., RNAi oligonucleotides) can be modified in various ways to improve or control specificity, stability, delivery, bioavailability, resistance to nuclease degradation, immunogenicity, base-pairing properties, RNA distribution and cellular uptake and other characteristics relevant to therapeutic research use. In some embodiments, the modification is a modified sugar. In some embodiments, the modification is a 5'-terminal phosphate group. In some embodiments, the modification is a modified internucleoside linkage. In some embodiments, the modification is a modified base. In some specific embodiments, the oligonucleotides described herein can comprise any one or any combination of the modifications described herein. For example, in some embodiments, oligonucleotides described herein comprise at least one modified sugar, a 5'-terminal phosphate group, at least one modified internucleoside linkage, and at least one modified sugar. Modified bases. The number of modifications on an oligonucleotide (eg, an RNAi oligonucleotide) and the location of those nucleotide modifications may affect the properties of the oligonucleotide. For example, oligonucleotides can be delivered in vivo by conjugating them to or including them in lipid nanoparticles (LNPs) or similar carriers. However, when the oligonucleotides are not protected by LNP or similar carriers, it may be advantageous to modify at least some of these nucleotides. Thus, in some embodiments, all or substantially all of the nucleotides of the oligonucleotide are modified. In some embodiments, more than half of the nucleotides are modified. In some embodiments, less than half of the nucleotides are modified. In some embodiments, the sugar moiety of all nucleotides comprising the oligonucleotide is modified at the 2' position. In some embodiments, the sugar moiety of all nucleotides of the oligonucleotide is at the 2' position, except for the nucleotide bound to the lipid (e.g., the 5'-terminal nucleotide of the sense strand). All have been modified. Modifications can be reversible or irreversible. In some embodiments, oligonucleotides as described herein possess sufficient properties to elicit desired properties (e.g., protection from enzymatic degradation, ability to target desired cells following in vivo administration, and/or thermodynamic stability) A certain number and type of modified nucleotides. Sugar modification In some embodiments, the nucleotide modifications in the sugar comprise 2'-modifications. In some specific embodiments, the 2'-modification can be 2'-O-propynyl, 2'-O-propylamino, 2'-amino, 2'-ethyl, 2'-fluoro(2 '-F), 2'-aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methoxyethyl (2'-MOE), 2'- O-[2-(methylamino)-2-side oxyethyl](2'-O-NMA), or 2'-deoxy-2'-fluoro-β-d-arabinose nucleic acid (2 '-FANA). In some embodiments, the modification is 2'-F, 2'-OMe or 2'-MOE. In some embodiments, the modification in the sugar includes modification of the sugar ring, which may include modification of one or more carbons of the sugar ring. For example, modification of the sugar of the nucleotide may include linking the 2'-oxygen of the sugar to the 1'-carbon or 4'-carbon of the sugar, or linking the 2'-oxygen to the 1' carbon via an ethyl or methylene bridge. -carbon or 4'-carbon linkage. In some embodiments, modified nucleotides have acyclic sugars that lack a 2'-carbon to 3'-carbon bond. In some embodiments, the modified nucleotide has, for example, a thiol group in the 4' position of the sugar. In some embodiments, lipid-bound RNAi oligonucleotides described herein comprise at least about 1 modified nucleotide (e.g., at least 1, at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60, or more). In some embodiments, the sense strand of a lipid-bound RNAi oligonucleotide comprises at least about 1 modified nucleotide (e.g., at least 1, at least 5, at least 10, at least 15, at least 20, at least 25 , at least 30, at least 35, or more). In some embodiments, the antisense strand of a lipid-bound RNAi oligonucleotide comprises at least about 1 modified nucleotide (e.g., at least 1, at least 5, at least 10, at least 15, at least 20, or more). In some embodiments, all nucleotides of the sense strand of the lipid-conjugated RNAi oligonucleotide are modified. In some embodiments, all nucleotides of the antisense strand of the lipid-conjugated RNAi oligonucleotide are modified. In some embodiments, all nucleotides of the lipid-bound RNAi oligonucleotide (ie, both the sense and antisense strands) are modified. In some embodiments, the modified nucleotides include 2'-modifications (e.g., 2'-F, or 2'-OMe, 2'-MOE, and 2'-deoxy-2'-fluoro-β -d-arabinose nucleic acid). In some embodiments, the present disclosure provides lipid-bound RNAi oligonucleotides with different modification patterns. In some embodiments, modified lipid-binding RNAi oligonucleotides comprise a sense strand sequence having a modification pattern as shown in the Examples and Sequence Listing and a modification pattern as shown in the Examples and Sequence Listing antisense sequence. In some embodiments, lipid-bound RNAi oligonucleotides disclosed herein comprise antisense strands having 2'-F modified nucleotides. In some embodiments, lipid-bound RNAi oligonucleotides disclosed herein comprise antisense strands comprising 2'-F and 2'-OMe modified nucleotides. In some embodiments, lipid-conjugated RNAi oligonucleotides disclosed herein comprise a sense strand having a 2'-F modified nucleotide. In some embodiments, lipid-bound RNAi oligonucleotides disclosed herein comprise a sense strand comprising nucleotides modified with 2'-F and 2'-OMe. In some embodiments, the oligonucleotides described herein comprise sense strands, wherein about 10 to 25%, 10%, 11%, 12%, 13%, 14%, 15%, 16% of the sense strands , 17%, 18%, 19%, or 20% of the nucleotides contained 2'-fluoro modifications. In some embodiments, about 11% of the nucleotides in the sense strand comprise 2-fluoro modifications. In some embodiments, about 20% of the nucleotides in the sense strand comprise 2-fluoro modifications. In some embodiments, the oligonucleotides described herein comprise antisense strands, wherein about 25 to 35%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, or 35% of the nucleotides contained 2'-fluoro modifications. In some embodiments, about 32% of the nucleotides of the antisense strand comprise 2'-fluoro modifications. In some specific embodiments, the oligonucleotide has about 15 to 25%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, or 25% % of its nucleotides contain 2'-fluoro modifications. In some embodiments, about 19% of the nucleotides in the oligonucleotide comprise 2'-fluoro modifications. In some embodiments, about 26% of the nucleotides in the oligonucleotide comprise 2'-fluoro modifications. In some embodiments, for such oligonucleotides, one or more of positions 8, 9, 10, or 11 of the sense strand is modified with a 2'-F group. In some embodiments, for such oligonucleotides, the sugar moiety at each nucleotide in the sense strand that is not modified with a 2'-F group or not bound to a lipid is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, and 12 to 20 in the sense strand is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 2 to 7, and 12 to 20 in the sense strand is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 6, and 12 to 20 in the sense strand is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 3, 5 to 7, and 12 to 20 in the sense strand is modified with 2'-OMe Grooming. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, and 13 to 20 in the sense strand is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, 12, and 14 to 20 in the sense strand is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, 12 to 17, and 19 to 20 in the sense strand is modified with 2'-OMe Grooming. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, and 12 to 19 in the sense strand is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, and 12 to 36 in the sense strand is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 2 to 7, and 12 to 36 in the sense strand is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 2 to 7, and 12 to 36 in the sense strand is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 3, 5 to 7, and 12 to 36 in the sense strand is modified with 2'-OMe Grooming. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, and 12 to 36 in the sense strand is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, and 13 to 36 in the sense strand is modified with 2'-OMe. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, and 12, and 14 to 36 in the sense strand is modified with 2'-OMe . In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, and 12 to 17, and 19 to 36 in the sense strand is via a 2'- OMe modified. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, and 12 to 19, and 21 to 36 in the sense strand is via a 2'- OMe modified. In some embodiments, for such oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, and 12 to 22, and 24 to 36 in the sense strand is via a 2'- OMe modified. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, 12 to 27, and 29 to 36 in the sense strand is modified with 2'-OMe Grooming. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, 12 to 28, and 30 to 36 in the sense strand is modified with 2'-OMe Grooming. In some embodiments, for these oligonucleotides, the sugar moiety at each nucleotide at positions 1 to 7, 12 to 29, and 31 to 36 in the sense strand is modified with 2'-OMe Grooming. In some embodiments, the sense strand comprises at least one 2'-F modified nucleotide, wherein the remaining nucleotides that are not modified with a 2'-F group or not bound to a lipid are modified with 2'-OMe. Grooming. In some embodiments, the antisense strand has 7 nucleotides modified with 2'-F at the 2' position of the sugar moiety. In some embodiments, the sugar moieties at positions 2, 3, 4, 5, 7, 10, and 14 of the antisense strand are modified with 2'-F. In some embodiments, the antisense strand has 14 nucleotides modified with 2'-OMe at the 2' position of the sugar moiety. In some embodiments, the sugar moieties at positions 6, 8, 9, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, and 22 of the antisense strand are via a 2'- OMe modified. In some embodiments, the sense strand has 4 nucleotides modified with 2'-F at the 2' position of the sugar moiety. In some embodiments, the sugar moieties at positions 2, 3, 8, 9, 10, and 11 of the sense strand are modified with 2'-F. In some embodiments, the sense strand has 15 nucleotides modified with 2'-OMe at the 2' position of the sugar moiety. In some embodiments, the sugar moieties at positions 6, 8, 9, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, and 22 of the antisense strand are via a 2'- OMe modified. In some embodiments, the antisense strand has 3 nucleotides modified with 2'-F at the 2'-position of the sugar moiety. In some embodiments, the sugar moieties of up to 3 nucleotides at positions 2, 5, and 14 of the antisense strand, and optionally at positions 1, 3, 7, and 10, are 2'-F modified . In some embodiments, the sugar moiety at each of positions 2, 5, and 14 of the antisense strand is modified with 2'-F. In other embodiments, the sugar moiety at each of positions 1, 2, 5, and 14 of the antisense strand is modified with 2'-F. In other embodiments, the sugar moiety at each of positions 2, 4, 5, and 14 of the antisense strand is modified with 2'-F. In some embodiments, the sugar moiety at each of positions 1, 2, 3, 5, 7, and 14 of the antisense strand is modified with 2'-F. In some embodiments, the sugar moiety at each of positions 2, 3, 4, 5, 7, and 14 of the antisense strand is modified with 2'-F. In some embodiments, the sugar moiety at each of positions 1, 2, 3, 5, 10, and 14 of the antisense strand is modified with 2'-F. In some embodiments, the sugar moiety at each of positions 2, 3, 4, 5, 10, and 14 of the antisense strand is modified with 2'-F. In some embodiments, the sugar moiety at each of positions 2, 3, 5, 7, 10, and 14 of the antisense strand is modified with 2'-F. In some embodiments, the sugar moiety at each of positions 2, 3, 4, 5, 7, 10, and 14 of the antisense strand is modified with 2'-F. In some embodiments, the antisense strand has 9 nucleotides modified with 2'-F at the 2'-position of the sugar moiety. In some embodiments, the sugar moiety at each of positions 2, 3, 4, 5, 7, 10, 14, 16, and 19 of the antisense strand is modified with 2'-F. In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise an antisense strand having a 2'- F-modified sugar moiety, and the sugar moiety of each remaining nucleotide of the antisense strand is modified by a modification selected from the group consisting of: 2'-O-propynyl, 2'-O-propylamino , 2'-amino, 2'-ethyl, 2'-aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methoxyethyl (2 '-MOE), 2'-O-[2-(methylamino)-2-side-oxyethyl](2'-O-NMA), and 2'-deoxy-2'-fluoro-β -d-arabinose nucleic acid (2'-FANA). In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise an antisense strand at positions 2, 3, 4, 5, 7, 10, 14, 16, and 19 of the antisense strand Each nucleotide at has a sugar moiety modified with 2'-F, and the sugar moiety of each remaining nucleotide of the antisense strand is modified with a modification selected from the group consisting of: 2'-O-propyne base, 2'-O-propylamino, 2'-amino, 2'-ethyl, 2'-aminoethyl (EA), 2'-O-methyl (2'-OMe), 2 '-O-methoxyethyl (2'-MOE), 2'-O-[2-(methylamino)-2-pentoxyethyl] (2'-O-NMA), and 2 '-Deoxy-2'-fluoro-β-d-arabinose nucleic acid (2'-FANA). In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise an antisense strand, each nucleotide at positions 1, 2, 5, and 14 of the antisense strand having a 2 '-F modified sugar moiety, and the sugar moiety of each remaining nucleotide of the antisense strand is modified by a modification selected from the group consisting of: 2'-O-propynyl, 2'-O-propyl Amino, 2'-amino, 2'-ethyl, 2'-aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methoxyethyl (2'-MOE), 2'-O-[2-(methylamino)-2-side-oxyethyl](2'-O-NMA), and 2'-deoxy-2'-fluoro -β-d-arabinose nucleic acid (2'-FANA). In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise an antisense strand with each nucleoside at positions 1, 2, 3, 5, 7, and 14 of the antisense strand. The acid has a sugar moiety modified with 2'-F, and the sugar moiety of each remaining nucleotide of the antisense strand is modified with a modification selected from the group consisting of: 2'-O-propynyl, 2'- O-propylamino, 2'-amino, 2'-ethyl, 2'-aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methyl Oxyethyl (2'-MOE), 2'-O-[2-(methylamino)-2-side oxyethyl] (2'-O-NMA), and 2'-deoxy- 2'-Fluoro-β-d-arabinose nucleic acid (2'-FANA). In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise an antisense strand with each nucleoside at positions 1, 2, 3, 5, 10, and 14 of the antisense strand. The acid has a sugar moiety modified with 2'-F, and the sugar moiety of each remaining nucleotide of the antisense strand is modified with a modification selected from the group consisting of: 2'-O-propynyl, 2'- O-propylamino, 2'-amino, 2'-ethyl, 2'-aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methyl Oxyethyl (2'-MOE), 2'-O-[2-(methylamino)-2-side oxyethyl] (2'-O-NMA), and 2'-deoxy- 2'-Fluoro-β-d-arabinose nucleic acid (2'-FANA). In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise an antisense strand with each nucleoside at positions 2, 3, 5, 7, 10, and 14 of the antisense strand. The acid has a sugar moiety modified with 2'-F, and the sugar moiety of each remaining nucleotide of the antisense strand is modified with a modification selected from the group consisting of: 2'-O-propynyl, 2'- O-propylamino, 2'-amino, 2'-ethyl, 2'-aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methyl Oxyethyl (2'-MOE), 2'-O-[2-(methylamino)-2-side oxyethyl] (2'-O-NMA), and 2'-deoxy- 2'-Fluoro-β-d-arabinose nucleic acid (2'-FANA). In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise an antisense strand at positions 2, 3, 4, 5, 7, 10, 14, 16, and Each nucleotide at position 19 has a sugar moiety modified with 2'-F, and the sugar moiety of each remaining nucleotide of the antisense strand is modified with a modification selected from the group consisting of: 2'-O-propanol Alkynyl, 2'-O-propylamino, 2'-amino, 2'-ethyl, 2'-aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methoxyethyl (2'-MOE), 2'-O-[2-(methylamino)-2-pentoxyethyl] (2'-O-NMA), and 2'-deoxy-2'-fluoro-β-d-arabinose nucleic acid (2'-FANA). In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise an antisense strand at each of positions 2, 3, 4, 5, 7, 10, and 14 of the antisense strand. The nucleotide has a sugar moiety modified with 2'-F, and the sugar moiety of each remaining nucleotide of the antisense strand is modified with a modification selected from the group consisting of: 2'-O-propynyl, 2 '-O-propylamino, 2'-amino, 2'-ethyl, 2'-aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O -Methoxyethyl (2'-MOE), 2'-O-[2-(methylamino)-2-side oxyethyl] (2'-O-NMA), and 2'-de Oxy-2'-fluoro-β-d-arabinose nucleic acid (2'-FANA). In some specific embodiments, lipid-bound RNAi oligonucleotides provided herein comprise antisense strands at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8. Position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, position 21, or position 22 has a longitude 2'- F modified sugar part. In some specific embodiments, lipid-bound RNAi oligonucleotides provided herein comprise antisense strands at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8. Position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, position 21, or position 22 has a longitude 2'- OMe modified sugar part. In some specific embodiments, lipid-bound RNAi oligonucleotides provided herein comprise antisense strands at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8. Position 9, Position 10, Position 11, Position 12, Position 13, Position 14, Position 15, Position 16, Position 17, Position 18, Position 19, Position 20, Position 21, or Position 22 has a member selected from the following Modified sugar moieties of the group consisting of: 2'-O-propynyl, 2'-O-propylamino, 2'-amino, 2'-ethyl, 2'-aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methoxyethyl (2'-MOE), 2'-O-[2-(methylamino)- 2-Pendant oxyethyl] (2'-O-NMA), and 2'-deoxy-2'-fluoro-β-d-arabinose nucleic acid (2'-FANA). In some specific embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise a sense strand having a 2'-F modified sugar moiety at positions 8 to 11. In some specific embodiments, lipid-bound RNAi oligonucleotides provided herein comprise a sense strand having a 2'-transition at positions 3, 5, 8, 10, 12, 13, 15, and 17 F modified sugar part. In some specific embodiments, lipid-bound RNAi oligonucleotides provided herein comprise a sense strand having a 2'OMe modified sugar at positions 1 to 7, and 12 to 17, or 12 to 20 part. In some specific embodiments, lipid-bound RNAi oligonucleotides provided herein comprise a sense strand having a 2'OMe modified sugar at positions 2 to 7, and 12 to 17, or 12 to 20 part. In some specific embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise a sense strand having a 2'OMe modified sugar moiety at positions 1 to 6, and 12 to 17, or 12 to 20 . In some specific embodiments, lipid-bound RNAi oligonucleotides provided herein comprise a sense strand at positions 1, 2, 4, 6, 7, 9, 11, 14, 16, and 18 to 20 There is a sugar moiety modified by 2'OMe. In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise a sense strand, each nucleotide at positions 1 to 7, and 12 to 17, or 12 to 20 of the sense strand has A sugar moiety modified with a modification selected from the group consisting of: 2'-O-propynyl, 2'-O-propylamino, 2'-amino, 2'-ethyl, 2'- Aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methoxyethyl (2'-MOE), 2'-O-[2-(methyl Amino)-2-Pendant oxyethyl] (2'-O-NMA), and 2'-deoxy-2'-fluoro-β-d-arabinose nucleic acid (2'-FANA). In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise a sense strand, wherein each nucleotide at positions 2 to 7, and 12 to 17, or 12 to 20 of the sense strand has A sugar moiety modified with a modification selected from the group consisting of: 2'-O-propynyl, 2'-O-propylamino, 2'-amino, 2'-ethyl, 2'- Aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methoxyethyl (2'-MOE), 2'-O-[2-(methyl Amino)-2-Pendant oxyethyl] (2'-O-NMA), and 2'-deoxy-2'-fluoro-β-d-arabinose nucleic acid (2'-FANA). In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise a sense strand, wherein each nucleotide at positions 1 to 6, and 12 to 17, or 12 to 20 of the sense strand has A sugar moiety modified with a modification selected from the group consisting of: 2'-O-propynyl, 2'-O-propylamino, 2'-amino, 2'-ethyl, 2'- Aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methoxyethyl (2'-MOE), 2'-O-[2-(methyl Amino)-2-Pendant oxyethyl] (2'-O-NMA), and 2'-deoxy-2'-fluoro-β-d-arabinose nucleic acid (2'-FANA). In some embodiments, lipid-bound RNAi oligonucleotides provided herein comprise a sense strand at positions 1, 2, 4, 6, 7, 9, 11, 14, 16, and Each nucleotide at positions 18 to 20 has a sugar moiety modified with a modification selected from the group consisting of: 2'-O-propynyl, 2'-O-propylamino, 2'-amino , 2'-ethyl, 2'-aminoethyl (EA), 2'-O-methyl (2'-OMe), 2'-O-methoxyethyl (2'-MOE), 2 '-O-[2-(methylamino)-2-side-oxyethyl](2'-O-NMA), and 2'-deoxy-2'-fluoro-β-d-arabinose nucleic acid (2'-FANA). In some specific embodiments, lipid-bound RNAi oligonucleotides provided herein comprise a sense strand at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8 , position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, position 21, position 22, position 23, position 24, position 25. Having a 2'-F modified sugar moiety at position 26, position 27, position 28, position 29, position 30, position 31, position 32, position 33, position 34, position 35, or position 36. In some specific embodiments, lipid-bound RNAi oligonucleotides provided herein comprise a sense strand at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8 , position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, position 21, position 22, position 23, position 24, position 25. Having a 2'-OMe modified sugar moiety at position 26, position 27, position 28, position 29, position 30, position 31, position 32, position 33, position 34, position 35, or position 36. In some specific embodiments, lipid-bound RNAi oligonucleotides provided herein comprise a sense strand at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8 , position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, position 21, position 22, position 23, position 24, position 25. Position 26, position 27, position 28, position 29, position 30, position 31, position 32, position 33, position 34, position 35, or position 36 having a modification selected from the group consisting of: Sugar part: 2'-O-propynyl, 2'-O-propylamino, 2'-amino, 2'-ethyl, 2'-aminoethyl (EA), 2'-O- Methyl (2'-OMe), 2'-O-methoxyethyl (2'-MOE), 2'-O-[2-(methylamino)-2-side oxyethyl]( 2'-O-NMA), and 2'-deoxy-2'-fluoro-β-d-arabinose nucleic acid (2'-FANA).5'- terminal phosphateIn some embodiments, lipid-conjugated RNAi oligonucleotides described herein comprise a 5'-terminal phosphate. In some embodiments, the 5'-terminal phosphate group of the lipid-conjugated RNAi oligonucleotide enhances the interaction with Ago2. However, oligonucleotides containing 5'-phosphate groups may be susceptible to degradation via phosphatases or other enzymes, which may limit their in vivo bioavailability. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise analogs of the 5'-phosphate that are resistant to such degradation. In some embodiments, the phosphate analog is oxymethylphosphonate, vinylphosphonate, or malonylphosphonate, or combinations thereof. In some embodiments, the 5' end of the lipid-conjugated RNAi oligonucleotide strand is attached to a chemical moiety that mimics the electrostatic and steric properties of the natural 5'-phosphate group ("phosphate mimetic"). mimic)”). In some embodiments, the lipid-conjugated RNAi oligonucleotides herein have a phosphate analog at the 4'-carbon position of the sugar (referred to as a "4'-phosphate analog"). )"). See, for example, International Patent Application Publication No. WO 2018/045317. In some embodiments, lipid-conjugated RNAi oligonucleotides herein comprise a 4'-phosphate analog at the 5'-terminal nucleotide. In some embodiments, the phosphate analog is an oxymethylphosphonate, wherein the oxygen atom of the oxymethyl group is bonded to a sugar moiety (eg, at its 4'-carbon) or an analog thereof. In other embodiments, the 4'-phosphate analog is a thiomethylphosphonate or an aminomethylphosphonate, wherein the sulfur atom of the thiomethyl group or the nitrogen atom of the aminomethyl group is with a sugar. Partial 4'-carbon or the like is joined. In some embodiments, the 4'-phosphate analog is oxymethylphosphonate. In some embodiments, the oxymethylphosphonate is of the formula -O-CH ₂-PO(OH) ₂,-O-CH ₂-PO(OR) ₂, or -O-CH2-POOH(R), where R is independently selected from H, CH ₃, alkyl, CH ₂CH ₂CN,CH ₂OCOC(CH ₃) ₃,CH ₂OCH ₂CH ₂Si(CH ₃) ₃, or protecting group. In some embodiments, the alkyl group is CH ₂CH ₃. More generally, R is independently selected from H, CH ₃or CH ₂CH ₃. In some specific embodiments, R is CH3. In some embodiments, the 4'-phosphate analog is 5'-methoxyphosphonate-4'-oxy. In some embodiments, the 4'-phosphate analog is 4'-oxymethylphosphonate. In some specific embodiments, lipid-conjugated RNAi oligonucleotides provided herein comprise: an antisense strand comprising a 4'-phosphate analog at the 5'-terminal nucleotide, wherein the 5'- The terminal nucleotide contains the following structure: Modified internucleotide linkagesIn some embodiments, lipid-conjugated RNAi oligonucleotides herein comprise modified internucleotide linkages. In some embodiments, the phosphate modification or substitution results in the oligonucleotide comprising at least about 1 (eg, at least 1, at least 2, at least 3, or at least 5) modified internucleotide linkages Union. In some specific embodiments, any of the oligonucleotides disclosed herein comprise about 1 to about 10 (e.g., 1 to 10, 2 to 8, 4 to 6, 3 to 10 , 5 to 10, 1 to 5, 1 to 3, or 1 to 2) modified inter-nucleotide linkages. In some specific embodiments, any of the oligonucleotide packages disclosed herein comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 modified nucleotides Inter-linkage. The modified inter-nucleotide linkage can be a phosphorodithioate linkage, a phosphorothioate linkage, a phosphotriester linkage, a thionoalkylphosphonate linkage, or a thionoalkylphosphonate linkage. Phosphonate triester linkage, phosphoamide linkage, phosphonate linkage, or borane phosphate linkage. In some specific embodiments, at least one modified internucleotide linkage of any of the oligonucleotides disclosed herein is a phosphorothioate linkage. In some embodiments, the lipid-bound RNAi oligonucleotides provided herein are at positions 1 and 2 of the sense strand, positions 1 and 2 of the antisense strand, positions 2 and 3 of the antisense strand, and There is a phosphorothioate linkage between one or more of positions 3 and 4 of the strand, positions 20 and 21 of the antisense strand, and positions 21 and 22 of the antisense strand. In some embodiments, the oligonucleotides described herein are at positions 1 and 2 of the sense strand, positions 1 and 2 of the antisense strand, positions 2 and 3 of the antisense strand, positions 20 and 2 of the antisense strand. 21, and each of positions 21 and 22 of the antisense strand has a phosphorothioate linkage. In some embodiments, the oligonucleotides described herein are at positions 1 and 2 of the sense strand, positions 1 and 2 of the antisense strand, positions 2 and 3 of the antisense strand, positions 3 and 3 of the antisense strand. 4. There is a phosphorothioate bond between positions 20 and 21 of the antisense strand and positions 21 and 22 of the antisense strand. In some specific embodiments, the oligonucleotides described herein are at positions 1 and 2 of the sense strand, positions 18 and 19 of the sense strand, positions 19 and 20 of the sense strand, positions 1 and 2 of the antisense strand, There is a phosphorothioate linkage between each of positions 2 and 3 of the antisense strand, positions 3 and 4 of the antisense strand, positions 20 and 21 of the antisense strand, and positions 21 and 22 of the antisense strand. In some embodiments, the oligonucleotide conjugates described herein comprise peptide nucleic acid (PNA). PNA is an oligonucleotide mimetic in which the sugar-phosphate backbone is replaced by a pseudopeptide skeleton composed of N-(2-aminoethyl)glycine units. The nucleobases are connected to this backbone via a two-atom carboxymethyl spacer. In some embodiments, the oligonucleotide conjugates described herein comprise N-morpholino oligomers (PMO), which contain methylene N linked through a diaminophosphate group. -The inter-nucleotide bonding backbone of the morpholine ring. base modificationIn some embodiments, lipid-conjugated RNAi oligonucleotides herein comprise one or more modified nucleobases. In some embodiments, a modified nucleobase (also referred to herein as a base analog) is attached at the 1' position of the sugar moiety of the nucleotide. In some embodiments, the modified nucleobase is a nitrogenous base. In some embodiments, the modified nucleobase contains no nitrogen atoms. See, for example, US Patent Application Publication No. 2008/0274462. In some embodiments, the modified nucleotide is a universal base. In some embodiments, the modified nucleotide contains no nucleobases (abase). In some embodiments, the universal base is a heterocyclic moiety located at the 1' position of the nucleotide sugar moiety in the modified nucleotide, or at the equivalent position in the substitution of the nucleotide sugar moiety, when When present in a duplex, the heterocyclic moiety can be positioned relative to more than one type of base without substantially changing the structure of the duplex. In some embodiments, a single-stranded nucleic acid containing universal bases forms a duplex with the target nucleic acid, which has A lower T than the duplex formed with the complementary nucleic acid _m. In some embodiments, the single-stranded nucleic acid containing the universal base forms a duplex with the target nucleic acid when compared to a reference single-stranded nucleic acid in which the universal base has been base-substituted to create a single mismatch. Strands have a higher T than duplexes formed from nucleic acids containing mismatches _m. Non-limiting examples of common conjugating nucleotides include, but are not limited to, inosine, 1-β-D-ribofuranosyl-5-nitroindole, and/or 1-β-D-ribofuranosyl -3-Nitropyrrole (see U.S. Patent Application Publication No. 2007/0254362; Van Aerschot et al.(1995)NUCLEIC ACIDS RES .23:4363-4370;Loakes et al.(1995)NUCLEIC ACIDS RES .23:2361-66; and Loakes & Brown (1994) NUCLEIC ACIDS RES. 22:4039-43). targeting ligand In some embodiments, it is desirable to target the oligonucleotides of the present disclosure (eg, lipid-conjugated RNAi oligonucleotides) to one or more cells or tissues of the central nervous system (CNS). Such a strategy may help avoid undesirable effects in other organs or avoid excessive depletion of the oligonucleotide to cells, tissues, or organs that do not benefit from the oligonucleotide. Accordingly, in some embodiments, lipid-binding RNAi oligonucleotides disclosed herein are modified to facilitate targeting and/or delivery to specific tissues, cells, or organs (e.g., to facilitate binding to delivery to CNS). In some embodiments, lipid-bound RNAi oligonucleotides comprise at least one (e.g., 1, 2, 3, 4, 5, 6, or more nucleotides) with one or more targeting partners. body-bound nucleotides. In some embodiments, one or more (e.g., 1, 2, 3, 4, 5, or 6) nucleotides of the lipid-conjugated RNAi oligonucleotides disclosed herein are each associated with a separate of targeted ligand binding. In some embodiments, one nucleotide of the lipid-conjugated RNAi oligonucleotides herein binds to a separate targeting ligand. In some embodiments, each of the 2 to 4 nucleotides of the lipid-conjugated RNAi oligonucleotides herein binds to a separate targeting ligand. In some embodiments, the targeting ligand is associated with 2 to 4 nucleotides at either end of the sense or antisense strand (e.g., the targeting ligand is associated with 5' or 3' of the sense or antisense strand). The 2 to 4 nucleotide overhangs or extensions on the 'end bind) so that the targeting ligand resembles the bristles of a toothbrush and the lipid-conjugated RNAi oligonucleotide resembles a toothbrush. For example, a lipid-conjugated RNAi oligonucleotide can contain a backbone-loop at the 5' or 3' end of the sense strand and 1, 2, 3, or 4 nucleotides of the backbone loop can be individually combined with Targeted ligand binding. In some embodiments, lipid-bound RNAi oligonucleotides provided by the present disclosure comprise a backbone-loop at the 3' end of the sense strand, wherein the loop of the backbone-loop contains three or four loops, And the 3 or 4 nucleotides containing three or four rings are individually combined with the targeting ligand. GalNAc is a high-affinity ligand for ASGPR, which is mainly expressed on the sinusoidal surface of hepatocytes and plays a role in binding, internalization, and subsequent clearance of circulating glycoproteins (asialoproteins) containing terminal galactose or GalNAc residues. Acid sugar protein) plays a major role. Binding (indirect or direct) of the GalNAc moiety to the oligonucleotides of the present disclosure can be used to target these oligonucleotides to ASGPR expressed on cells. In some embodiments, oligonucleotides of the disclosure bind to at least one or more GalNAc moieties, wherein the GalNAc moiety targets the oligonucleotide to ASGPR expressed on human liver cells (eg, human hepatocytes). In some embodiments, the GalNAc moiety targets the oligonucleotide to the liver. In some embodiments, the oligonucleotides of the present disclosure bind to monovalent GalNAc, either directly or indirectly. In some embodiments, the oligonucleotide binds, directly or indirectly, to more than one monovalent GalNAc (i.e., to 2, 3, or 4 monovalent GalNAc moieties, and typically to 3, or 4 monovalent GalNAc moieties). GalNAc partially binds) binds. In some embodiments, the oligonucleotide is conjugated to one or more bivalent GalNAc, trivalent GalNAc, or tetravalent GalNAc moieties. In some embodiments, one or more (eg, 1, 2, 3, 4, 5, or 6) nucleotides of the oligonucleotide are each bound to the GalNAc moiety. In some embodiments, 2 to 4 nucleotides of the four loops each bind to a separate GalNAc. In some embodiments, 1 to 3 nucleotides of the tricycle are each bound to a separate GalNAc. In some embodiments, the targeting ligand is bound to 2 to 4 nucleotides at either end of the sense or antisense strand (e.g., the ligand is bound to the 5' or 3' end of the sense or antisense strand). The 2 to 4 nucleotide overhangs or extensions on the oligonucleotide bind) so that the GalNAc portion resembles the bristles of a toothbrush and the oligonucleotide resembles a toothbrush. In some embodiments, the GalNAc moiety is bound to the nucleotide of the sense strand. For example, four (4) GalNAc moieties may bind to nucleotides in the four loops of the sense strand, with each GalNAc moiety binding to 1 nucleotide. In some embodiments, any combination of tetracyclic adenine and guanine nucleotides. In some specific embodiments, tetracycle (L) has a monovalent GalNAc moiety attached to any one or more guanine nucleotides of the tetracycle via any linker described herein, as shown below (X = hetero atom): In some specific embodiments, tetracycle (L) has a monovalent GalNAc moiety attached to any one or more adenine nucleotides of the tetracycle via any linker described herein, as shown below (X = hetero atom): In some embodiments, the lipid-conjugated RNAi oligonucleotides herein comprise a monovalent GalNAc attached to a guanine nucleotide, referred to as [ademG-GalNAc] or 2'-aminodiethoxy Methanol-guanine-GalNAc, as shown below: In some embodiments, lipid-conjugated RNAi oligonucleotides herein comprise a monovalent GalNAc attached to an adenine nucleotide, referred to as [ademA-GalNAc] or 2'-aminodiethoxy Methanol-Adenine-GalNAc, as shown below: An example of such a conjugate comprising a loop of the 5' to 3' nucleotide sequence GAAA (L=linker, X=heteroatom) is shown below. Such loops may be present, for example, at positions 27 to 30 of the righteous strand. In the chemical formula, is used to describe the point of attachment to an oligonucleotide strand. Appropriate methods or chemistry (eg, click chemistry) can be used to attach the targeting ligand to the nucleotide. In some embodiments, the targeting ligand binds to the nucleotide using a click linker. In some embodiments, an acetal-based linker may be used to conjugate a targeting ligand to the nucleotide of any of the oligonucleotides described herein. Acetal-based linkers are disclosed, for example, in International Patent Application Publication No. WO 2016/100401. In some embodiments, the linker is an unstable linker. However, in other embodiments, the linker is stable. Shown below are examples of loops containing the 5' to 3' nucleotide GAAA, where the GalNAc moiety is attached to the nucleotides of the loop using an acetal linker. Such loops may be present, for example, at positions 27 to 30 of the righteous strand. In the chemical formula, It is the attachment point to the oligonucleotide strand. As described above, targeting ligands can be attached to nucleotides using a variety of suitable methods or chemical synthesis techniques (eg, click chemistry). In some embodiments, the targeting ligand binds to the nucleotide using a click linker. In some embodiments, an acetal-based linker may be used to conjugate a targeting ligand to the nucleotide of any of the oligonucleotides described herein. Acetal-based linkers are disclosed, for example, in International Patent Application Publication No. WO 2016/100401. In some embodiments, the linker is an unstable linker. However, in other embodiments, the linker is a stable linker. In some embodiments, a double-stranded extension (e.g., having up to 3, 4, 5, or 6 bp in length) is provided between the targeting ligand (e.g., GalNAc moiety) and the lipid-bound RNAi oligonucleotide. ). In some embodiments, the lipid-bound RNAi oligonucleotides herein do not have GalNAc bound thereto. Lipid conjugates In some embodiments, any of the lipid moieties described herein binds to the nucleotides of the sense strand of the oligonucleotide. In some embodiments, the lipid moiety is bound to a terminal position of the oligonucleotide. In some embodiments, the lipid moiety is bound to the 5' terminal nucleotide of the sense strand. In some embodiments, the lipid moiety is bound to the 3' terminal nucleotide of the sense strand. In some embodiments, the lipid moiety binds to internal nucleotides on the sense strand. An internal position is any nucleotide position other than the two terminal positions at each end of the sense strand. In some embodiments, the lipid moiety is bound to one or more internal positions of the sense strand. In some embodiments, the lipid moiety is associated with position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8, position 9, position 10, position 11, position 12, Location 13, location 14, location 15, location 16, location 17, location 18, location 19, location 20, location 21, location 22, location 23, location 24, location 25, location 26, location 27, location 28, location 29 , position 30, position 31, position 32, position 33, position 34, position 35, or position 36 combined. In some embodiments, the lipid moiety binds to position 1 of the sense strand. In some embodiments, the lipid moiety binds to position 4 of the sense strand. In some embodiments, the lipid moiety binds to position 8 of the sense strand. In some embodiments, the lipid moiety binds to position 12 of the sense strand. In some embodiments, the lipid moiety binds to position 13 of the sense strand. In some embodiments, the lipid moiety binds to position 18 of the sense strand. In some embodiments, the lipid moiety binds to position 20 of the sense strand. In some embodiments, the lipid moiety binds to position 23 of the sense strand. In some embodiments, the lipid moiety binds to position 28 of the sense strand. In some embodiments, the lipid moiety binds to position 29 of the sense strand. In some embodiments, the lipid moiety binds to position 30 of the sense strand. In some embodiments, lipid-binding RNAi oligonucleotides described herein comprise at least one nucleotide that binds to one or more lipid moieties. In some embodiments, one or more lipid moieties are bound to the same nucleotide. In some embodiments, one or more lipid moieties are bound to different nucleotides. In some embodiments, one, two, three, four, five, or six lipid moieties are conjugated to the oligonucleotide. In some embodiments, one or more lipid moieties are bound to adenine nucleotides. In some embodiments, one or more lipid moieties are conjugated to guanine nucleotides. In some embodiments, one or more lipid moieties are bound to cytosine nucleotides. In some embodiments, one or more lipid moieties are bound to thymine nucleotides. In some embodiments, one or more lipid moieties are conjugated to uracil nucleotides. In some embodiments, the lipid moiety is a hydrocarbon chain. In some embodiments, the hydrocarbon chain is saturated. In some embodiments, the hydrocarbon chain is unsaturated. In some embodiments, the hydrocarbon chain is branched. In some embodiments, the hydrocarbon chain is straight. In some embodiments, the lipid moiety is a C8 to C30 hydrocarbon chain. In some specific embodiments, the lipid moiety is C8:0, C10:0, C11:0, C12:0, C14:0, C16:0, C17:0, C18:0, C18:1, C18:2, C22:5, C22:0, C24:0, C26:0, C22:6, C24:1, diyl C16:0 or diyl C18:1. In some embodiments, the lipid moiety is a C16 hydrocarbon chain. In some embodiments, the lipid moiety is conjugated to the oligonucleotide via a linker. In some embodiments, the nucleotide of the lipid-conjugated oligonucleotide is represented by Formula II-b or II-c: or a pharmaceutically acceptable salt thereof, wherein: L ¹It is a covalent bond, monovalent or divalent saturated or unsaturated, linear or branched C _1-50Hydrocarbon chain, in which 0 to 10 methylene units of the hydrocarbon chain are independently replaced by: -Cy-, -O-, -C(O)NR-, -NR-, -S-, -C( O)-, -C(O)O-, -S(O)-, -S(O) ₂-, -P(O)OR-, -P(S)OR-, or ; R ⁴System hydrogen, R ^A, or a suitable amine protecting group; and R ⁵It is adamantyl, or saturated or unsaturated, straight chain, or branched C _1-50Hydrocarbon chain, in which 0 to 10 methylene units of the hydrocarbon chain are independently replaced by: -O-, -C(O)NR-, -NR-, -S-, -C(O)-, -C(O)O-, -S(O)-, -S(O) ₂-, -P(O)OR-, or -P(S)OR. In some embodiments of lipid-conjugated RNAi oligonucleotides, R ⁵Department selected from In certain embodiments of lipid-conjugated RNAi oligonucleotides, R ⁵Department selected from In some embodiments, R ⁵department In some embodiments, R ⁵department In some embodiments, the nucleotide of the lipid-conjugated RNAi oligonucleotide is represented by the formula II-Ibor II-IcRepresents: or a pharmaceutically acceptable salt thereof; wherein B series nucleobase, or hydrogen; m ranges from 1 to 50; X ¹Department -O-, or -S-; Y series hydrogen, ,or ; R ³It is hydrogen or a suitable protecting group; X ²Department O, or S; X ³is -O-, -S-, or covalent bond; Y ¹A linking group attached to the 2'- or 3'-end of a nucleoside, nucleotide, or oligonucleotide; Y ²Be hydrogen, a phosphoramidite analog, an internucleotide linking group attached to the 5'-end of a nucleoside, nucleotide, or oligonucleotide, or a linking group attached to a solid support ; R ⁵It is adamantyl, or saturated or unsaturated, straight chain, or branched C _1-50Hydrocarbon chain, in which 0 to 10 methylene units of the hydrocarbon chain are independently replaced by: -O-, -C(O)NR-, -NR-, -S-, -C(O)-, -C(O)O-, -S(O)-, -S(O) ₂-, -P(O)OR-, or -P(S)OR-; and R is hydrogen, a suitable protecting group, or optionally a substituted group selected from: C _1-6Aliphatic, phenyl, 4 to 7 membered saturated or partially unsaturated heterocyclic ring with 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 1 to 4 heteroatoms independently selected from nitrogen, oxygen , and 5- to 6-membered heteroaryl rings of heteroatoms of sulfur. In some embodiments, the lipid system In some embodiments, the oligonucleotides of the oligonucleotide-ligand conjugates are double-stranded molecules. In some embodiments, the oligonucleotides are RNAi molecules. In some embodiments, the double-stranded oligonucleotide contains backbone loops. In some embodiments, the backbone loop is shown as S1-L-S2, where S1 is complementary to S2, and where L forms a loop between S1 and S2. In some embodiments, the ligand binds to any of the nucleotides in the loops of the backbone loops. In some embodiments, the ligand binds to any of the nucleotides in the backbone of the backbone loop. In some embodiments, the ligand binds to the first nucleotide from 5' to 3' of the loop. In some embodiments, the ligand binds to the second nucleotide from 5' to 3' of the loop. In some embodiments, the ligand binds to the third nucleotide from 5' to 3' of the loop. In some embodiments, the ligand binds to the fourth nucleotide from 5' to 3' in the loop. In some embodiments, the ligand binds to one, two, three, or four nucleotides in the loop. In some embodiments, the ligand binds to three nucleotides in the backbone loop. In some embodiments, the backbone loop is 16 nucleotides long. In some embodiments, the ligand binds to the third nucleotide from 5' to 3' of the backbone loop. In some embodiments, the ligand binds to the eighth nucleotide from 5' to 3' in the backbone loop. In some embodiments, the ligand binds to the ninth nucleotide from 5' to 3' in the backbone loop. In some embodiments, the ligand binds to the tenth nucleotide from 5' to 3' in the backbone loop. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a 20 nucleotide sense strand, with positions numbered 1 to 20 from 5' to 3'. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a lipid bound to position 1 of the 20 nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a lipid bound to position 4 of the 20 nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a lipid bound to position 8 of the 20 nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a lipid bound to position 12 of the 20 nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a lipid bound to position 13 of the 20 nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a lipid bound to position 18 of the 20 nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a lipid bound to position 20 of the 20 nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a 36 nucleotide sense strand, with positions numbered 1 to 36 from 5' to 3'. In some embodiments, the lipid-bound RNAi oligonucleotide-comprises a lipid bound to position 1 of the 36-nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide-comprises a lipid bound to position 4 of the 36-nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide-comprises a lipid bound to position 8 of the 36-nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide-comprises a lipid bound to position 12 of the 36-nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide-comprises a lipid bound to position 13 of the 36-nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide-comprises a lipid bound to position 18 of the 36-nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide-comprises a lipid bound to position 20 of the 36-nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide-comprises a lipid bound to position 23 of the 36-nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide-comprises a lipid bound to position 28 of the 36-nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide-comprises lipid binding to position 29 of the 36-nucleotide sense strand. In some embodiments, the lipid-bound RNAi oligonucleotide-comprises a lipid bound to position 30 of the 36-nucleotide sense strand. Exemplary oligonucleotides In some embodiments, lipid-conjugated RNAi oligonucleotides comprise nucleotides conjugated to fatty acids. In some embodiments, the fatty acid is a saturated fatty acid. In some embodiments, the fatty acid is an unsaturated fatty acid. In some embodiments, lipid-bound RNAi oligonucleotides comprise nucleotides that bind to lipids. In some embodiments, the lipid is a carbon chain. In some embodiments, the carbon chain is saturated. In some embodiments, the carbon chain is unsaturated. In some embodiments, lipid-bound RNAi oligonucleotides comprise nucleotides bound to a 16 carbon (C16) lipid. In some specific embodiments, the C16 lipid contains at least one double bond. In some embodiments, the oligonucleotide of the lipid-conjugated RNAi oligonucleotide is conjugated to a C16 lipid as follows: In some embodiments, lipid-bound RNAi oligonucleotides comprise a 20 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotide comprises a 22 nucleotide long antisense strand. In some embodiments, the sense strand is 20 nucleotides long and the antisense strand is 22 nucleotides long. In some embodiments, the lipid-conjugated RNAi oligonucleotide comprises a 20 nucleotide long sense strand and a 22 nucleotide long antisense strand, wherein the sense strand and antisense strand form 20 bases For the double-stranded area. In some embodiments, the 3' end of the sense strand is blunt. In some embodiments, the 5' end of the antisense strand is blunt. In some embodiments, the 3' end of the antisense strand includes an overhang. In some embodiments, the overhang is 2 nucleotides long. In some embodiments, the overhang is GG. In some embodiments, lipid-bound RNAi oligonucleotides comprise one or more 2' modifications. In some embodiments, the 2' modification is selected from 2'-fluoro or 2'-methyl. In some embodiments, lipid-bound RNAi oligonucleotides comprise an antisense strand and a sense strand as described herein, wherein the sense strand comprises at least one hydrocarbon chain bound to the 5' terminal nucleotide of the sense strand. In some embodiments, lipid-bound RNAi oligonucleotides comprise an antisense strand and a sense strand as described herein, wherein the sense strand comprises at least one C16 hydrocarbon chain bound to the 5' terminal nucleotide of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide comprises a 22- to 24-nucleotide antisense strand and a 20- to 22-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, wherein the sense strand contains at least one hydrocarbon chain bound to the 5' terminal nucleotide of the sense strand. In some embodiments, a lipid-bound RNAi oligonucleotide comprises a 22- to 24-nucleotide antisense strand and a 20- to 22-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, wherein the sense strand contains at least one C16 hydrocarbon chain bound to the 5' terminal nucleotide of the sense strand. In some embodiments, a lipid-bound RNAi oligonucleotide comprises a 22- to 24-nucleotide antisense strand and a 20- to 22-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a 20 to 22 base pair asymmetric double-stranded region with an overhang on the 3' end of the antisense strand and a blunt end on the 3' end of the oligonucleotide, The sense strand includes at least one hydrocarbon chain bound to the 5' terminal nucleotide on the sense strand. In some embodiments, a lipid-bound RNAi oligonucleotide comprises a 22- to 24-nucleotide antisense strand and a 20- to 22-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a 20 to 22 base pair asymmetric double-stranded region with an overhang on the 3' end of the antisense strand and a blunt end on the 3' end of the oligonucleotide, The sense strand includes at least one C16 hydrocarbon chain bound to the 5' terminal nucleotide on the sense strand. In some embodiments, a lipid-bound RNAi oligonucleotide comprises an antisense strand and a sense strand as described herein, wherein the sense strand comprises at least one nucleotide internal to the sense strand (e.g., at position 7 of nucleotides) combined with a hydrocarbon chain. In some embodiments, a lipid-bound RNAi oligonucleotide comprises an antisense strand and a sense strand as described herein, wherein the sense strand comprises at least one nucleotide internal to the sense strand (e.g., at position 7 nucleotide) combined C16 hydrocarbon chain. In some embodiments, not all internal nucleotides are suitable for lipid binding to stellate cells that deliver RNAi oligonucleotides to the CNS. For example, in some embodiments, conjugates at positions 9 or 10 on the sense strand, numbered from 5' to 3', are not suitable for delivery of RNAi oligonucleotides to stellate cells in the CNS. In some embodiments, lipid conjugates at internal positions of the sense strand numbered from 5' to 3' do not include positions 9 and 10. In some embodiments, a lipid-bound RNAi oligonucleotide comprises a 22- to 24-nucleotide antisense strand and a 20- to 22-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, wherein the sense strand contains at least one hydrocarbon chain bound to an internal nucleotide of the sense strand (eg, the nucleotide at position 7). In some embodiments, a lipid-bound RNAi oligonucleotide comprises a 22- to 24-nucleotide antisense strand and a 20- to 22-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, wherein the sense strand contains at least one C16 hydrocarbon chain bound to an internal nucleotide of the sense strand (eg, the nucleotide at position 7). In some embodiments, a lipid-bound RNAi oligonucleotide comprises a 22- to 24-nucleotide antisense strand and a 20- to 22-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a 20 to 22 base pair asymmetric double-stranded region with an overhang on the 3' end of the antisense strand and a blunt end on the 3' end of the oligonucleotide, The sense strand includes at least one hydrocarbon chain bound to an internal nucleotide of the sense strand (eg, the nucleotide at position 7). In some embodiments, a lipid-bound RNAi oligonucleotide comprises a 22- to 24-nucleotide antisense strand and a 20- to 22-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a 20 to 22 base pair asymmetric double-stranded region with an overhang on the 3' end of the antisense strand and a blunt end on the 3' end of the oligonucleotide, The sense strand includes at least one C16 hydrocarbon chain bound to an internal nucleotide of the sense strand (eg, the nucleotide at position 7). In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotides, [ademX-Ls] = lipid attached to nucleotides with phosphorothioate linkages to adjacent nucleotides, and [ademX -L]=lipid attached to nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotide, and [ademX-L] = lipid attached to the nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotide, and [ademX-L] = lipid attached to the nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotide, [ademX-L] = lipid attached to the nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotide, and [ademX-L] = lipid attached to the nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotide, and [ademX-L] = lipid attached to the nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotide, and [ademX-L] = lipid attached to the nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, the lipid-conjugated RNAi oligonucleotide comprises a 36 nucleotide long sense strand. In some embodiments, the lipid-conjugated RNAi oligonucleotide comprises a 22 nucleotide long antisense strand. In some embodiments, the sense strand is 36 nucleotides long and the antisense strand is 22 nucleotides long. In some embodiments, the lipid-bound RNAi oligonucleotide comprises a 36 nucleotide long sense strand and a 22 nucleotide long antisense strand, wherein the sense strand and antisense strand form 20 bases For the double-stranded area. In some embodiments, the 3' end of the sense strand contains a backbone-loop. In some embodiments, the 3' end of the sense strand contains four rings. In some embodiments, the 3' end of the sense strand comprises: a stem-loop comprising SEQ ID NO: 32. In some embodiments, the 3' end of the antisense strand includes an overhang. In some embodiments, the overhang is 2 nucleotides long. In some embodiments, the overhang is GG. In some embodiments, a lipid-bound RNAi oligonucleotide comprises a sense strand comprising a backbone-loop at its 3' end and at least one hydrocarbon chain bound to the 5' terminal nucleotide of the sense strand. In some embodiments, a lipid-bound RNAi oligonucleotide comprises a sense strand comprising a backbone-loop at its 3' end and at least one hydrocarbon chain bound to the nucleotide of the sense strand. In some embodiments, a lipid-bound RNAi oligonucleotide comprises a sense strand comprising a backbone-loop at its 3' end and at least one C16 hydrocarbon strand bound to the 5' terminal nucleotide of the sense strand. In some embodiments, lipid-bound RNAi oligonucleotides comprise a sense strand comprising a tetracyclic loop and at least one C16 hydrocarbon chain bound to a nucleotide of the tetracyclic loop. In some embodiments, lipid-bound RNAi oligonucleotides comprise a 22- to 24-nucleotide antisense strand and a 20- to 36-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, in which the sense strand contains at least one C16 hydrocarbon chain bound to the first nucleotide of the sense strand (position 1 from 5'>3'). In some embodiments, lipid-bound RNAi oligonucleotides comprise a 22- to 24-nucleotide antisense strand and a 20- to 36-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, in which the sense strand contains at least one C16 hydrocarbon chain bound to the fourth nucleotide (position 4 from 5'>3') of the sense strand. In some embodiments, lipid-bound RNAi oligonucleotides comprise a 22- to 24-nucleotide antisense strand and a 20- to 36-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, in which the sense strand contains at least one C16 hydrocarbon chain bound to the eighth nucleotide (position 8 from 5'>3') of the sense strand. In some embodiments, lipid-bound RNAi oligonucleotides comprise a 22- to 24-nucleotide antisense strand and a 20- to 36-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, in which the sense strand contains at least one C16 hydrocarbon chain bound to the twelfth nucleotide (position 12 from 5'>3') of the sense strand. In some embodiments, lipid-bound RNAi oligonucleotides comprise a 22- to 24-nucleotide antisense strand and a 20- to 36-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, in which the sense strand contains at least one C16 hydrocarbon chain bound to the thirteenth nucleotide (position 13 from 5'>3') of the sense strand. In some embodiments, lipid-bound RNAi oligonucleotides comprise a 22- to 24-nucleotide antisense strand and a 20- to 36-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, in which the sense strand contains at least one C16 hydrocarbon chain bound to the eighteenth nucleotide (position 18 from 5'>3') of the sense strand. In some embodiments, lipid-bound RNAi oligonucleotides comprise a 22- to 24-nucleotide antisense strand and a 20- to 36-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, in which the sense strand contains at least one C16 hydrocarbon chain bound to the twentieth nucleotide (position 20 from 5'>3') of the sense strand. In some embodiments, lipid-bound RNAi oligonucleotides comprise a 22- to 24-nucleotide antisense strand and a 20- to 36-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, in which the sense strand contains at least one C16 hydrocarbon chain bound to the twenty-third nucleotide (position 23 from 5'>3') of the sense strand. In some embodiments, lipid-bound RNAi oligonucleotides comprise a 22- to 24-nucleotide antisense strand and a 20- to 36-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, in which the sense strand contains at least one C16 hydrocarbon chain bound to the twenty-eighth nucleotide (position 28 from 5'>3') of the sense strand. In some embodiments, lipid-bound RNAi oligonucleotides comprise a 22- to 24-nucleotide antisense strand and a 20- to 36-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, in which the sense strand contains at least one C16 hydrocarbon chain bound to the twenty-ninth nucleotide (position 29 from 5'>3') of the sense strand. In some embodiments, lipid-bound RNAi oligonucleotides comprise a 22- to 24-nucleotide antisense strand and a 20- to 36-nucleotide sense strand as described herein, wherein the antisense strand and The sense strand forms a double-stranded region of 20 to 22 base pairs, in which the sense strand contains at least one C16 hydrocarbon chain bound to the thirtieth nucleotide (position 30 from 5'>3') of the sense strand. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O-Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotides, [ademX-Ls] = lipid attached to nucleotides with phosphorothioate linkages to adjacent nucleotides, and [ademX -L]=lipid attached to nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotides, [ademX-Ls] = lipid attached to nucleotides with phosphorothioate linkages to adjacent nucleotides, and [ademX -L]=lipid attached to nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotides, [ademX-Ls] = lipid attached to nucleotides with phosphorothioate linkages to adjacent nucleotides, and [ademX -L]=lipid attached to nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotides, [ademX-Ls] = lipid attached to nucleotides with phosphorothioate linkages to adjacent nucleotides, and [ademX -L]=lipid attached to nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotides, [ademX-Ls] = lipid attached to nucleotides with phosphorothioate linkages to adjacent nucleotides, and [ademX -L]=lipid attached to nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotides, [ademX-Ls] = lipid attached to nucleotides with phosphorothioate linkages to adjacent nucleotides, and [ademX -L]=lipid attached to nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotides, [ademX-Ls] = lipid attached to nucleotides with phosphorothioate linkages to adjacent nucleotides, and [ademX -L]=lipid attached to nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotides, [ademX-Ls] = lipid attached to nucleotides with phosphorothioate linkages to adjacent nucleotides, and [ademX -L]=lipid attached to nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotide, and [ademX-L] = lipid attached to the nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O- Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotide, and [ademX-L] = lipid attached to the nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell target genes comprise the following modification pattern Cross to: Where [mXs] = 2'- phosphorothioate linkage to adjacent nucleotide O- Methyl modified nucleotide, [fXs] = 2'-fluoro modified nucleotide with phosphorothioate linkage to adjacent nucleotide, [mX] = phosphodiester with adjacent nucleotide The Sutra of Key Links 2'- O - Methyl modified nucleotide, [fX] = 2'-fluoro modified nucleotide with phosphodiester linkage to adjacent nucleotide, [MePhosphonate-4O-mX] = 4'-O- Monomethylphosphonate-2'-O-methyl modified nucleotides, [ademX-Ls] = lipid attached to nucleotides with phosphorothioate linkages to adjacent nucleotides, and [ademX -L]=lipid attached to nucleotide, optionally where L is a C16 hydrocarbon. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell mRNA in the spinal cord comprise a sense strand comprising a backbone-loop at its 3' end and at least one strand associated with the sense strand The hydrocarbon chain of the nucleotide binding at position 2, position 3, position 6, position 13, position 14, position 15, position 19, position 20, position 23, position 28, position 29, or position 30, where the position is Numbered from 5' to 3'. In some embodiments, a lipid-conjugated RNAi oligonucleotide for reducing the expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a backbone-loop at its 3' end and at least one linkage with the sense strand. A hydrocarbon chain bound to a nucleotide at position 1, position 4, position 8, position 12, position 13, position 18, position 20, position 23, position 28, position 29, or position 30, where the position is from 5' to 3' number. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell mRNA in the cerebellum comprise a sense strand comprising a backbone-loop at its 3' end and at least one covalent linkage with the sense strand. A hydrocarbon chain bound to a nucleotide at position 1, position 4, position 23, position 28, position 29, or position 30, where the positions are numbered from 5' to 3'. In some embodiments, a lipid-conjugated RNAi oligonucleotide for reducing the expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a backbone-loop at its 3' end and at least one covalent linkage with the sense strand. A hydrocarbon chain bound to a nucleotide at position 1, position 4, position 12, position 13, position 18, position 20, position 23, position 28, position 29, or position 30, where the position is from 5' to 3' number. In some embodiments, a lipid-conjugated RNAi oligonucleotide for reducing expression of stellate cell mRNA in the frontal cortex comprises a sense strand comprising a stem-loop at its 3' end and at least one The hydrocarbon chain of the nucleotide bound at position 23 of the sense strand, where the positions are numbered from 5' to 3'. In some embodiments, a lipid-conjugated RNAi oligonucleotide for reducing the expression of stellate cell mRNA in the spinal cord comprises a sense strand having a blunt end at its 3' end and at least one position corresponding to the sense strand. 1. A hydrocarbon chain bound to a nucleotide at position 4, position 8, position 12, position 13, position 18, or position 20, where the positions are numbered from 5' to 3'. In some embodiments, a lipid-conjugated RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a blunt-ended sense strand at its 3' end and at least one position corresponding to the sense strand. 1. A hydrocarbon chain bound to a nucleotide at position 4, position 8, position 12, position 13, position 18, or position 20, where the positions are numbered from 5' to 3'. In some embodiments, a lipid-conjugated RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a blunt end at its 3' end and at least one position corresponding to the sense strand. 4. A hydrocarbon chain bound to a nucleotide at position 12, position 13, position 18, or position 20, where the positions are numbered from 5' to 3'. In some embodiments, lipid-conjugated RNAi oligonucleotides for reducing expression of stellate cell mRNA in the hypothalamus comprise a sense strand with a blunt end at its 3' end and at least one position corresponding to the sense strand. 1. A hydrocarbon chain bound to a nucleotide at position 4, position 12, position 13, position 18, or position 20, where the positions are numbered from 5' to 3'. In some embodiments, a lipid-conjugated RNAi oligonucleotide for reducing the expression of stellate cell mRNA in the frontal cortex comprises a sense strand comprising a blunt end at its 3' end and at least one covalent linkage with the sense strand. A hydrocarbon chain bound to a nucleotide at position 4, where the positions are numbered from 5' to 3'. In some embodiments, position numbering throughout this description is based on numbering from 5' to 3', such that the terminal nucleotide at the 5' end is position 1. In some embodiments, lipid-bound RNAi oligonucleotides for reducing expression of stellate cell mRNA in the spinal cord comprise a sense strand comprising a stem-loop at its 3' end and binding to position 1 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the spinal cord comprises a sense strand comprising a stem-loop at its 3' end and binding to position 4 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the spinal cord comprises a sense strand comprising a stem-loop at its 3' end and binding to position 8 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the spinal cord comprises a sense strand comprising a stem-loop at its 3' end and binding to position 12 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the spinal cord comprises a sense strand comprising a stem-loop at its 3' end and binding to position 13 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the spinal cord comprises a sense strand comprising a stem-loop at its 3' end and binding to position 18 of the sense strand of lipids. In some embodiments, lipid-bound RNAi oligonucleotides for reducing expression of stellate cell mRNA in the spinal cord comprise a sense strand comprising a stem-loop at its 3' end and binding to position 20 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the spinal cord comprises a sense strand comprising a stem-loop at its 3' end and binding to position 23 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the spinal cord comprises a sense strand comprising a stem-loop at its 3' end and binding to position 28 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the spinal cord comprises a sense strand comprising a stem-loop at its 3' end and binding to position 29 of the sense strand of lipids. In some embodiments, lipid-bound RNAi oligonucleotides for reducing expression of stellate cell mRNA in the spinal cord comprise a sense strand comprising a stem-loop at its 3' end and binding to position 30 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata includes at its 3' end the sense strand of the backbone-loop and binds to position 1 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata includes at its 3' end the sense strand of the backbone-loop and binds to position 4 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata includes at its 3' end the sense strand of the backbone-loop and binds to position 18 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata includes at its 3' end the sense strand of the backbone-loop and binds to position 19 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 20 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 23 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 28 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 29 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 30 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a stem-loop at its 3' end and binding to position 1 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a stem-loop at its 3' end and binding to position 4 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a stem-loop at its 3' end and binding to position 23 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a stem-loop at its 3' end and binding to position 28 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a stem-loop at its 3' end and binding to position 29 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises at its 3' end the sense strand of the backbone-loop and binding to position 1 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises at its 3' end the sense strand of the backbone-loop and binding to position 4 of the sense strand. Lipids. In some embodiments, lipid-bound RNAi oligonucleotides for reducing expression of stellate cell mRNA in the hypothalamus comprise a backbone-looped sense strand at its 3' end and binding to position 12 of the sense strand. Lipids. In some embodiments, lipid-bound RNAi oligonucleotides for reducing expression of stellate cell mRNA in the hypothalamus comprise a backbone-looped sense strand at its 3' end and binding to position 13 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 18 of the sense strand of lipids. In some embodiments, lipid-bound RNAi oligonucleotides for reducing expression of stellate cell mRNA in the hypothalamus comprise a sense strand comprising a stem-loop at its 3' end and binding to position 20 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 23 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 28 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 29 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 30 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the frontal cortex comprises a sense strand comprising a stem-loop at its 3' end and a position relative to the sense strand 23-bound lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a backbone-loop at its 3' end and position 1 of the sense strand A conjugated lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a backbone-loop at its 3' end and position 4 relative to the sense strand A conjugated lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a backbone-loop at its 3' end and position 8 relative to the sense strand A conjugated lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a backbone-loop at its 3' end and position 12 relative to the sense strand A conjugated lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a backbone-loop at its 3' end and position 13 relative to the sense strand A conjugated lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a backbone-loop at its 3' end and position 18 relative to the sense strand A conjugated lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a backbone-loop at its 3' end and position 20 relative to the sense strand A conjugated lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a backbone-loop at its 3' end and position 23 relative to the sense strand A conjugated lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a backbone-loop at its 3' end and position 28 relative to the sense strand A conjugated lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a backbone-loop at its 3' end and position 29 relative to the sense strand A conjugated lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a backbone-loop at its 3' end and position 30 relative to the sense strand A conjugated lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 1 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 4 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 8 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 12 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 13 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 18 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 20 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 23 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 28 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 29 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a stem-loop at its 3' end and binding to position 30 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a stem-loop at its 3' end and binding to position 1 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a stem-loop at its 3' end and binding to position 4 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a stem-loop at its 3' end and binding to position 23 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a stem-loop at its 3' end and binding to position 28 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a stem-loop at its 3' end and binding to position 29 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, lipid-bound RNAi oligonucleotides for reducing expression of stellate cell mRNA in the hypothalamus comprise a sense strand comprising a stem-loop at its 3' end and binding to position 1 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 4 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 12 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 13 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 18 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, lipid-bound RNAi oligonucleotides for reducing expression of stellate cell mRNA in the hypothalamus comprise a sense strand comprising a stem-loop at its 3' end and binding to position 20 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 23 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 28 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 29 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a stem-loop at its 3' end and binding to position 30 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the frontal cortex comprises a sense strand comprising a stem-loop at its 3' end and a position relative to the sense strand 23-bound lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand containing a blunt end at its 3' end and binding to position 1 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a blunt end at its 3' end and binding to position 4 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing the expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand containing a blunt end at the 3' end of the sense strand and binding to position 8 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a blunt end at its 3' end and binding to position 12 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand containing a blunt end at its 3' end and binding to position 13 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a blunt end at its 3' end and binding to position 18 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand containing a blunt end at its 3' end and binding to position 20 of the sense strand. Lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 1 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 4 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 8 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 12 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 13 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 18 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 20 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 4 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 12 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 13 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 18 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 20 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 1 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 4 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 12 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 13 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 18 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 20 of the sense strand . In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the frontal cortex comprises a sense strand containing a blunt end at its 3' end and binding to position 4 of the sense strand of lipids. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand containing a blunt end at its 3' end and binding to position 1 of the sense strand. Lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a blunt end at its 3' end and binding to position 4 of the sense strand. Lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a blunt end at its 3' end and binding to position 8 of the sense strand. Lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a blunt end at its 3' end and binding to position 12 of the sense strand. Lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord includes a blunt-ended sense strand at its 3' end and a lipid binding to position 13 of the sense strand. , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand comprising a blunt end at its 3' end and binding to position 18 of the sense strand. Lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the lumbar spinal cord comprises a sense strand containing a blunt end at its 3' end and binding to position 20 of the sense strand. Lipid, and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 1 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 4 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 8 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 12 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 13 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 18 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the medulla oblongata comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 20 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 4 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 12 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 13 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 18 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the cerebellum comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 20 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 1 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 4 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 12 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 13 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 18 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the hypothalamus comprises a sense strand comprising a blunt end at its 3' end and a lipid binding to position 20 of the sense strand , and wherein the oligonucleotide comprises at least one modified nucleotide. In some embodiments, a lipid-bound RNAi oligonucleotide for reducing expression of stellate cell mRNA in the frontal cortex comprises a sense strand containing a blunt end at its 3' end and binding to position 4 of the sense strand A lipid, and wherein the oligonucleotide comprises at least one modified nucleotide.Provides general methods for nucleic acids and their analogsNucleic acids and analogs comprising the lipid conjugates described herein can be prepared using a variety of methods known in the art, including standard phosphoramidite methods. Any phosphoramidite synthesis method can be used to synthesize the nucleic acids provided in this disclosure. In certain embodiments, phosphoramidites are used in solid phase synthesis methods to produce reactive intermediate phosphite compounds, which are subsequently oxidized using known methods to produce phosphonate modified oligonucleosides Acids, typically with phosphodiester or phosphorothioate internucleotide linkages. Oligonucleotide synthesis of the present disclosure can be performed in either direction: from 5' to 3' or from 3' to 5' using methods known in the art. In certain embodiments, methods for synthesizing provided nucleic acids comprise (a) attaching a nucleoside or analog thereof to a solid support via a covalent linkage; (b) attaching a nucleoside phosphoramidite or The analog thereof is coupled to the reactive hydroxyl group on the nucleoside or analog thereof of step (a) to form an internucleotide bond therebetween, wherein any uncoupled nucleoside or analog thereof on the solid support is used capping with a capping reagent; (c) oxidizing the internucleotide bond with an oxidizing agent; and (d) repeating steps (b) to (c) with a subsequent nucleoside phosphoramidite or analog thereof to form a nucleic acid or Analogs thereof, wherein at least the nucleoside of step (a) or an analog thereof, the nucleoside phosphoramidite of step (b) or an analog thereof, or the subsequent nucleoside phosphoramidite of step (d) or an analog thereof At least one of them includes a lipid conjugate moiety described herein. Generally, the coupling, capping/oxidation steps, and optional deprotection steps are repeated until the oligonucleotide reaches the desired length and/or sequence, after which it is cleaved from the solid support. In certain embodiments, oligonucleotides are prepared that include 1 to 3 nucleic acids or analogs thereof that comprise lipid conjugate units on a tetracyclic ring. below process AWhere specific protecting groups, leaving groups, or transformation conditions are shown, one of ordinary skill in the art will understand that other protecting groups, leaving groups, and transformation conditions are also suitable and may be considered. Also consider process ACertain reactive functional groups requiring additional protecting group strategies (eg, -N(H)-, -OH, etc.) are contemplated and understood by those of ordinary skill in the art. Such groups and transformations are described in detail in March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, M. B. Smith and J. March, 5 ^thEdition, John Wiley & Sons, 2001. COMPREHENSIVE ORGANIC TRANSFORMATIONS,(R. C. Larock, 2 ^ndEdition, John Wiley & Sons, 1999), and PROTECTING GROUPS IN ORGANIC SYNTHESIS,(T. W. Greene and P. G. M. Wuts, 3 ^rdedition, John Wiley & Sons, 1999), each of which is hereby incorporated by reference in its entirety. In certain embodiments, the nucleic acids of the present disclosure, and their analogs, are substantially in accordance with the process A ,process A1,and process BPreparation: process A :Synthesis of ligand-binding oligonucleotides disclosed herein process A1 :Synthesis of the disclosed lipid-binding oligonucleotides As above process Aand process A1As shown in , the formula I-1Nucleic acids or analogs thereof are combined with one or more ligands/lipophilic compounds to form a formula containing a plurality of ligands/lipid conjugates Ior formula Iaof compounds. Generally speaking, the combination is performed in series or parallel by techniques known in the art. I-1or formula I-1aThe esterification or amidation reaction between nucleic acids or analogs thereof and one or more adamantyl and/or lipophilic compounds (eg, fatty acids) is carried out. Then the formula can be Ior formula Iadeprotect the nucleic acid or its analogues to form the formula I-2or formula I-2acompound and protected with a suitable hydroxyl protecting group (e.g., DMTr) to form the formula I-3or formula I-3aof compounds. In an aspect, the formula I-3or formula I-3aThe nucleic acid-ligand conjugates can be covalently attached to a solid support (e.g., via a succinic acid linking group) to form a formula containing one or more adamantyl and/or lipid conjugates. I-4or formula I-4aSolid support nucleic acid-ligand conjugate or the like. In another aspect, the formula I-3or formula I-3aNucleic acid-ligand conjugates can form reagents with P(III) (e.g., 2-cyanoethyl N, N-Di-isopropyl phosphoridite chloride) reacts to form a formula containing a P(III) group I-5or formula I-5anucleic acid or its analogues. Then, the formula can be I-5or formula I-5aThe nucleic acid-ligand conjugate or analog thereof is subjected to oligopolymerization conditions using known and commonly used processes to prepare oligonucleotides in the art. For example, consider the formula I-5or formula I-5aThe compound is coupled to a solid supported nucleic acid-ligand conjugate bearing a 5'-hydroxyl group or an analog thereof. Further steps may include one or more deprotection, coupling, phosphite oxidation, and/or cleavage from the solid support to provide oligonucleotides of various nucleotide lengths represented by the formula II-1or formula II-IaCompounds represented include one or more lipid conjugate nucleotide units. B,E,L,ligand,LC,n,PG ¹,PG ²,PG ⁴,R ¹,R ²,R ³,X,X ¹,X ²,X ³Each of , and Z is as defined above and described herein. Process B: Synthesis of lipid conjugates after oligonucleotides of the present disclosure As above process BAs shown in , the formula can be I-1Nucleic acid or its analogues are deprotected to form the formula I-6The compound is protected with a suitable hydroxyl protecting group (e.g., DMTr) to form the formula I-7compounds and form reagents with P(III) (e.g. 2-cyanoethyl N, N-Di-isopropyl phosphoridite chloride) reacts to form a formula containing a P(III) group I-8nucleic acid or its analogues. Next, the formula can be I-8The nucleic acid or analog thereof is subjected to oligopolymerization conditions using known and commonly used processes to prepare oligonucleotides in the art. For example, consider the formula I-8The compound is coupled to a solid supported nucleic acid bearing a 5'-hydroxyl group or an analog thereof. Further steps may include one or more deprotection, coupling, phosphite oxidation, and/or cleavage from the solid support to provide the formula II-2The compounds represent oligonucleotides of various nucleotide lengths. Then the formula can be II-2The oligonucleotide is combined with one or more ligands (e.g., adamantyl, or a lipophilic compound (e.g., fatty acid)) to form a formula containing one or more ligand conjugates II-1of compounds. Generally speaking, the combination is performed in series or parallel by techniques known in the art. II-2It is carried out by esterification or amidation reaction between nucleic acid or its analog and one or more adamantyl groups or fatty acids. B,E,L,ligand,LC,n,PG ¹,PG ²,PG ⁴,R ¹,R ²,R ³,X,X ¹,X ²,X ³Each of , and Z is as defined above and described herein. In certain embodiments, the nucleic acids of the present disclosure, and analogs thereof, are formulated according to the methods shown below. process Cand process DPreparation: Procedure C: Synthesis of Lipid-Conjugated Oligonucleotides of the Disclosure As above process CAs shown in , the formula C1Nucleic acid or its analogues are protected to form the formula C2of compounds. Then change the formula C2Alkylation of nucleic acids or analogs thereof (e.g., via Pummerer rearrangement using DMSO and acetic acid) to form formula C3Monothioacetal compounds. Next, change the formula C3Nucleic acid or its analogues and C4Couple under appropriate conditions (e.g., mild oxidizing conditions) to form the formula C5nucleic acid or its analogues. Then the formula can be C5deprotect the nucleic acid or its analogues to form the formula C6compound, and with the formula C7The ligand (adamantyl or lipophilic compound (e.g., fatty acid)) is coupled under appropriate amide-forming conditions (e.g., HATU, DIPEA) to form a formula comprising a lipid conjugate of the present disclosure IbNucleic acid-ligand conjugates or analogs thereof. Then the formula can be IbThe nucleic acid-ligand conjugate or its analog is deprotected to form the formula C8compound and protected with a suitable hydroxyl protecting group (e.g., DMTr) to form the formula C9of compounds. In an aspect, the formula C9The nucleic acid, or analog thereof, can be covalently attached to a solid support (e.g., via a succinic acid linker) to form a formula containing the ligand conjugates (adamantyl and/or lipid moieties) of the present disclosure. C10Solid support nucleic acid-ligand conjugate or the like. In another aspect, the formula C9Nucleic acid-ligand conjugates or analogs thereof can form reagents with P(III) (e.g., 2-cyanoethyl N, N-Di-isopropyl phosphoridite chloride) reacts to form a formula containing a P(III) group C11Nucleic acid-ligand conjugates or analogs thereof. Then, the formula can be C11The nucleic acid-ligand conjugate or analog thereof is subjected to oligopolymerization conditions using known and commonly used processes to prepare oligonucleotides in the art. For example, consider the formula C11The compound is coupled to a solid supported nucleic acid-ligand conjugate bearing a 5'-hydroxyl group or an analog thereof. Further steps may include one or more deprotection, coupling, phosphite oxidation, and/or cleavage from the solid support to provide oligonucleotides of various nucleotide lengths represented by the formula II-b-3Compounds represented include one or more adamantyl and/or lipid-binding nucleotide units. B,E,L ²,PG ¹,PG ²,PG ³,PG ⁴,R ¹,R ²,R ³,R ⁴,R ⁵,X ¹,X ²,X ³Each of , V, W, and Z is as defined above and described herein. Process D: Synthesis of lipid conjugates after oligonucleotides of the present disclosure B,E,L ²,PG ¹,PG ²,PG ³,PG ⁴,R ¹,R ²,R ³,R ⁴,R ⁵,X ¹,X ²,X ³Each of , V, W, and Z is as defined above and described herein. As above process DAs shown in , the formula can be C5Nucleic acids or analogs thereof are selectively deprotected to form the formula D1The compound is protected with a suitable hydroxyl protecting group (e.g., DMTr) to form the formula D2compounds and form reagents with P(III) (e.g. 2-cyanoethyl N, N-di-isopropyl phosphoramidite chloride) reacts to form the formula D3nucleic acid or its analogues. Next, change the formula D3The nucleic acid or its analogue is subjected to pre-prepared oligopolymerization conditions using known and commonly used processes to prepare oligonucleotides in the art. For example, consider the formula D3The compound is coupled to a solid supported nucleic acid bearing a 5'-hydroxyl group or an analog thereof. Further steps may include one or more deprotection, coupling, phosphite oxidation, and/or cleavage from the solid support to provide the formula D4The compounds represent oligonucleotides of various nucleotide lengths. Then the formula can be D4The oligonucleotide is deprotected to form the formula D5compounds and coupled with hydrophobic ligands (e.g., adamantyl or lipophilic moieties) to form C7(e.g., adamantyl or fatty acid) compounds are coupled under appropriate amide-forming conditions (e.g., HATU, DIPEA) to form formulas comprising conjugates of the disclosed ligands (e.g., adamantyl or fatty acid) II-b-3of oligonucleotides. One of ordinary skill in the art will understand that various functional groups (such as aliphatic groups, alcohols, carboxylic acids, esters, amides, aldehydes, halogens, and nitriles) present in the nucleic acids of the present disclosure or their analogs can be Interconversion by techniques well known in the art, including but not limited to reduction, oxidation, esterification, hydrolysis, partial oxidation, partial reduction, halogenation, dehydration, partial hydration, and hydration. See e.g., " MARCH'S ADVANCED ORGANIC CHEMISTRY",(5 ^thEd., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001), each of which is incorporated by reference in its entirety. Such interconversion may require one or more of the aforementioned techniques, and certain methods for synthesizing the nucleic acids provided by this disclosure are described in the examples below. In some embodiments, the present disclosure provides methods for preparing oligonucleotides comprising one or more lipid conjugates, or pharmaceutically acceptable salts thereof, the lipid conjugate units being represented by the formula II-a-1Represents: This method includes the following steps: (a) Offering I-5aNucleic acid or its analogues: or its salt, and (b) general formula I-5aThe compound oligomericly polymerizes to form the formula II-1acompound, among which B,E,L,LC,n,PG ⁴,R ¹,R ²,R ³,X,X ¹,X ²,X ³Each of , E, and Z is as defined above and described herein. In the above step (b), oligopolymerization refers to using known and commonly used processes to perform oligopolymerization forming conditions to prepare oligonucleotides in the art. For example, consider the formula I-5aThe compound is coupled to a solid supported nucleic acid bearing a 5'-hydroxyl group or an analog thereof. Further steps may include one or more deprotection, coupling, phosphite oxidation, and cleavage from the solid support to provide the lipid conjugate-containing formula of the present disclosure. II-1aThe compounds represent oligonucleotides of various nucleotide lengths. In some embodiments, the present disclosure provides methods for preparing oligonucleotides comprising one or more lipid conjugates, further comprising preparing I-5aNucleic acid or its analogues: or its salt, the method includes the following steps: (a) Offering IaNucleic acid or its analogues: or its salt, (b) general formula IaThe nucleic acid or analog thereof is deprotected to form the formula I-2aThe compound of: or its salt, (c) general formula I-2The nucleic acid or its analog is protected to form the formula I-3aThe compound of: or its salt, and (d) general formula I-3aThe nucleic acid or analog thereof is treated with a P(III) forming reagent to form the formula I-5aNucleic acid or its analogues, wherein B,E,L,LC,n,PG ⁴,R ¹,R ²,R ³,X,X ¹,X ²,X ³Each of , E, and Z is as defined above and described herein. In step (b) above, the formula IaThe compound of PG ¹and PG ²Contains silyl ethers or cyclic silene derivatives, which can be removed under acidic conditions or with fluoride anions. Examples of reagents that provide fluoride anions to remove silicon-based protecting groups include hydrofluoric acid, pyridine hydrogen fluoride, triethylamine trihydrofuride, tetrafluorofluoride N-Butylammonium, etc. In step (c) above, the formula I-2aThe compounds are protected with appropriate hydroxyl protecting groups. In some embodiments, for protective I-2aThe protective group PG of the 5'-hydroxyl group of the compound ⁴Includes acid-labile protecting groups such as trityl, 4-methoxytrityl, 4,4'-dimethoxytrityl, 4,4',4"-trimethoxytrityl base, 9-phenyl-𠮿-9-yl (9-phenyl-xanthen-9-yl), 9-(p-phenylmethyl)-𠮿-9-yl, phenyl𠮿yl (pixyl), 2,7 - dimethylphenyl hydroxyl group, etc. In certain embodiments, the acid-labile protecting group is suitable for use during both solution and solid phase synthesis of acid-sensitive nucleic acids or the like using, for example, dichloroacetic acid or trichloroacetic acid to remove protection. In the above step (d), the equation I-3aThe compound is treated with a P(III)-forming reagent to obtain the formula I-5aof compounds. In the context of this disclosure, a P(III)-forming reagent is a phosphorus reagent that reacts to form a phosphorus(III) compound. In some embodiments, the P(III) forming reagent is 2-cyanoethyl N, N-Diisopropyl phosphoridite chloride or 2-cyanoethyl dichlorophosphate. In certain embodiments, the P(III) forming reagent is 2-cyanoethyl N, N-Diisopropylphosphoridite chloride. Those with ordinary knowledge should realize that in the form I-3aCompound X ¹Displacement of the leaving group in the P(III)-forming reagent is accomplished in the presence or absence of a suitable base. Such suitable bases are well known in the art and include organic and inorganic bases. In certain embodiments, the basic tertiary amine is such as triethylamine or diisopropylethylamine. In other embodiments, step (d) above uses N, N- Dimethylphosphoramidite dichloride is carried out as P(V) forming reagent. In some embodiments, the present disclosure provides methods for preparing oligonucleotides comprising one or more lipid conjugates, further comprising preparing IaNucleic acid-lipid conjugates or analogs thereof: or its salt, the method includes the following steps: (a) Offering I-1Nucleic acid or its analogues: or its salt, and, (b) Combine one or more lipophilic compounds with the formula I-1Nucleic acids or analogs thereof are combined to form a formula comprising one or more lipid conjugates IaNucleic acids or analogs thereof, wherein: B,E,L,LC,n,PG ¹,PG ²,R1,R ²,X,X ¹Each of , and Z is as defined above and described herein. In step (b) above, the formula I-1aNucleic acids or analogs thereof are combined with one or more lipophilic compounds to form a formula comprising one or more lipid conjugates of the present disclosure. Iaof compounds. Generally speaking, the combination is performed in series or parallel by techniques known in the art. I-1aIt is carried out by an esterification or amidation reaction between a nucleic acid or its analog and one or more fatty acids. In certain embodiments, conjugation is performed under suitable amide-forming conditions to provide a formula comprising a plurality of lipid conjugates. Iof compounds. Suitable amide forming conditions may include the use of amide coupling agents known in the art, such as, but not limited to, HATU, PyBOP, DCC, DIC, EDC, HBTU, HCTU, PyAOP, PyBrOP, BOP, BOP-Cl, DEPBT, T3P, TATU, TBTU, TNTU, TOTU, TPTU, TSTU, or TDBTU. Alternatively, the binding of lipophilic compounds can be achieved by surface AThis can be accomplished by any of the cross-coupling techniques described in . In some embodiments, the present disclosure provides methods for preparing pharmaceutically acceptable salts of oligonucleotides comprising one or more lipid conjugates, the lipid conjugate units being represented by the formula II-1Represents: This method includes the following steps: (a) Offering II-2The oligonucleotide: or its salt, and, (b) Combine one or more lipophilic compounds with the formula II-2oligonucleotides are combined to form a formula containing one or more lipid conjugates II-1of oligonucleotides. In step (b) above, the formula II-2The oligonucleotide is combined with one or more lipophilic compounds to form a formula including one or more lipid conjugates of the present disclosure. II-1of oligonucleotides. Generally speaking, the combination is performed in series or parallel by techniques known in the art. II-2It is carried out by esterification or amidation reaction between oligonucleotide and one or more fatty acids. In certain embodiments, conjugation is performed under suitable amide-forming conditions to provide a formula comprising a plurality of lipid conjugates. II-1Oligonucleotides. Suitable amide forming conditions may include the use of amide coupling agents known in the art, such as, but not limited to, HATU, PyBOP, DCC, DIC, EDC, HBTU, HCTU, PyAOP, PyBrOP, BOP, BOP-Cl, DEPBT, T3P, TATU, TBTU, TNTU, TOTU, TPTU, TSTU, or TDBTU. Alternatively, the binding of lipophilic compounds can be achieved by surface AThis can be accomplished by any of the cross-coupling techniques described in . In some embodiments, the present disclosure provides for preparation of a compound comprising: II-2Oligonucleotide of the unit represented: Or its pharmaceutically acceptable salt method, which includes the following steps: (a) Offering I-8Nucleic acid or its analogues: or its salt, and (b) general formula I-8The compound oligomericly polymerizes to form the formula II-2of compounds. In the above step (b), oligopolymerization refers to using known and commonly used processes to perform oligopolymerization forming conditions to prepare oligonucleotides in the art. For example, consider the formula I-8The compound is coupled to a solid supported nucleic acid bearing a 5'-hydroxyl group or an analog thereof. Further steps may include one or more deprotection, coupling, phosphite oxidation, and cleavage from the solid support to provide Eq. II-2The compounds represent oligonucleotides of various nucleotide lengths. In some embodiments, the present disclosure provides methods for preparing nucleic acids or analogs thereof comprising one or more lipid conjugates, further comprising preparing a formula I-8Nucleic acid or its analogues: or its salt, the method includes the following steps: (a) Offering I-1Nucleic acid or its analogues: or its salt, (b) general formula I-1The nucleic acid or analog thereof is deprotected to form the formula I-6The compound of: or its salt, (c) general formula I-6The nucleic acid or its analog is protected to form the formula I-7The compound of: or its salt, and (d) general formula I-7The nucleic acid or analog thereof is treated with a P(III) forming reagent to form the formula I-8Nucleic acid or analog thereof, in step (b) above, the formula I-1The compound of PG ¹and PG ²Contains silyl ethers or cyclic silene derivatives, which can be removed under acidic conditions or with fluoride anions. Examples of reagents that provide fluoride anions to remove silicon-based protecting groups include hydrofluoric acid, pyridine hydrogen fluoride, triethylamine trihydrofuride, tetrafluorofluoride N-Butylammonium, etc. In step (c) above, the formula I-6The compounds are protected with appropriate hydroxyl protecting groups. In some embodiments, for protective I-6The protective group PG of the 5'-hydroxyl group of the compound ⁴Includes acid-labile protecting groups such as trityl, 4-methoxytrityl, 4,4'-dimethoxytrityl, 4,4',4"-trimethoxytrityl base, 9-phenyl-𠮿-9-yl, 9-(p-phenylmethyl)-𠮿-9-yl, phenyl𠮿yl, 2,7-dimethylphenyl𠮿yl, etc. In some specific In embodiments, acid-labile protecting groups are suitable for deprotection using, for example, dichloroacetic acid or trichloroacetic acid during both solution and solid phase synthesis of acid-sensitive nucleic acids or analogs thereof. In the above step (d), the equation I-7The compound is treated with a P(III)-forming reagent to obtain the formula I-8of compounds. In the context of this disclosure, a P(III)-forming reagent is a phosphorus reagent that reacts to form a phosphorus(III) compound. In some embodiments, the P(III) forming reagent is 2-cyanoethyl N, N-Diisopropyl phosphoridite chloride or 2-cyanoethyl dichlorophosphate. In certain embodiments, the P(III) forming reagent is 2-cyanoethyl N, N-Diisopropylphosphoridite chloride. Those with ordinary knowledge should realize that in the form I-7Compound X ¹Displacement of the leaving group in the P(III)-forming reagent is accomplished in the presence or absence of a suitable base. Such suitable bases are well known in the art and include organic and inorganic bases. In certain embodiments, the basic tertiary amine is such as triethylamine or diisopropylethylamine. In other embodiments, step (d) above uses N, N- Dimethylphosphoramidite dichloride is carried out as P(V) forming reagent. In some embodiments, the present disclosure provides methods for preparing oligonucleotide-ligand conjugates comprising one or more adamantyl and/or lipid moieties, or pharmaceutically acceptable salts thereof, the lipid conjugates The object unit consists of the formula II-b-3Represents: This method includes the following steps: (a) Offering C11Nucleic acid-ligand conjugates or analogs thereof: or its salt, and (b) general formula C11The compound oligomericly polymerizes to form the formula II-b-3of compounds. In the above step (b), oligopolymerization refers to using known and commonly used processes to perform oligopolymerization forming conditions to prepare oligonucleotides in the art. For example, consider the formula C11The compound is coupled to a solid supported nucleic acid bearing a 5'-hydroxyl group or an analog thereof. Further steps may include one or more deprotection, coupling, phosphite oxidation, and cleavage from the solid support to provide oligos of various nucleotide lengths with one or more nucleic acid-ligand conjugate units. Glycoside-ligand conjugates, wherein each unit is composed of a formula containing an adamantyl or lipid moiety of the present disclosure II-b-3represented by the compound. In some embodiments, formulas for preparing one or more lipid conjugates II-b-3The method of oligonucleotide further comprises preparing formula C11Nucleic acid-lipid conjugates or analogs thereof: or its salt, the method includes the following steps: (a) Offering ibNucleic acid or its analogues: or its salt, (b) general formula IbThe nucleic acid-ligand conjugate or analog thereof is deprotected to form the formula C8The compound of: or its salt, (c) general formula C8The nucleic acid-ligand conjugate or analog thereof is protected to form the formula C9The compound of: or its salt, and (d) general formula C9The nucleic acid-ligand conjugate or analog thereof is treated with a P(III) forming reagent to form the formula C11nucleic acid or its analogues. In step (b) above, the formula IbThe compound of PG ¹and PG ²Contains silyl ethers or cyclic silene derivatives, which can be removed under acidic conditions or with fluoride anions. Examples of reagents that provide fluoride anions to remove silicon-based protecting groups include hydrofluoric acid, pyridine hydrogen fluoride, triethylamine trihydrofuride, tetrafluorofluoride N-Butylammonium, etc. In step (c) above, the formula C8The compounds are protected with appropriate hydroxyl protecting groups. In some embodiments, for protective C8The protective group PG of the 5'-hydroxyl group of the compound ⁴Includes acid-labile protecting groups such as trityl, 4-methoxytrityl, 4,4'-dimethoxytrityl, 4,4',4"-trimethoxytrityl base, 9-phenyl-𠮿-9-yl, 9-(p-phenylmethyl)-𠮿-9-yl, phenyl𠮿yl, 2,7-dimethylphenyl𠮿yl, etc. In some specific In embodiments, acid-labile protecting groups are suitable for deprotection using, for example, dichloroacetic acid or trichloroacetic acid during both solution and solid phase synthesis of acid-sensitive nucleic acids or analogs thereof. In the above step (d), the equation C9The compound is treated with a P(III)-forming reagent to obtain the formula C11of compounds. In the context of this disclosure, a P(III)-forming reagent is a phosphorus reagent that reacts to form a phosphorus(III) compound. In some embodiments, the P(III) forming reagent is 2-cyanoethyl N, N-Diisopropyl phosphoridite chloride or 2-cyanoethyl dichlorophosphate. In certain embodiments, the P(III) forming reagent is 2-cyanoethyl N, N-Diisopropylphosphoridite chloride. Those with ordinary knowledge should realize that in the form C9Compound X ¹Displacement of the leaving group in the P(III)-forming reagent is accomplished in the presence or absence of a suitable base. Such suitable bases are well known in the art and include organic and inorganic bases. In certain embodiments, the basic tertiary amine is such as triethylamine or diisopropylethylamine. In other embodiments, step (d) above uses N, N- Dimethylphosphoramidite dichloride is carried out as P(V) forming reagent. In some embodiments, the present disclosure provides for preparing formula II-b-3A method of oligonucleotide-ligand conjugates, the oligonucleotide-ligand conjugates comprising one or more nucleic acid-ligand conjugate units each containing one or more adamantyl or lipid moieties, the The method further includes the preparation formula IbNucleic acid-ligand conjugates or analogs thereof: or its salt, the method includes the following steps: (a) Offering C6Nucleic acid-ligand conjugates or analogs thereof: or its salt, and, (b) Combine the lipophilic compound with the formula C6Nucleic acids or analogs thereof are combined to form a formula containing one or more adamantyl and/or lipid conjugates IbNucleic acid-ligand conjugates or analogs thereof. In step (b) above, the conjugation is carried out under suitable amide formation conditions to obtain a formula containing an adamantyl and/or lipid conjugate. ibof compounds. Suitable amide forming conditions may include the use of amide coupling agents known in the art, such as, but not limited to, HATU, PyBOP, DCC, DIC, EDC, HBTU, HCTU, PyAOP, PyBrOP, BOP, BOP-Cl, DEPBT, T3P, TATU, TBTU, TNTU, TOTU, TPTU, TSTU, or TDBTU. In certain embodiments, the amide forming conditions include HATU, and DIPEA, or TEA. In certain embodiments, the formula C6The nucleic acid-ligand conjugate or analog thereof is provided in salt form (eg, fumarate) and first converted to the free base (eg, using sodium bicarbonate) before performing the conjugation step. In some embodiments, the present disclosure provides for preparing formula II-b-3A method for oligonucleotide-ligand conjugates, the oligonucleotide-ligand conjugates comprising one or more nucleic acid-ligand conjugate units, the method further comprising the preparation formula C6Nucleic acid-ligand conjugates or analogs thereof: or its salt, the method includes the following steps: (a) Offering C1Nucleic acid or its analogues: or its salt, and, (b) general formula C1The nucleic acid or its analog is protected to form the formula C2The compound of: or its salt, (c) general formula C2The nucleic acid or analog thereof is alkylated to form the formula C3The compound of: or its salt, (d) general formula C3The nucleic acid or its analog is of the formula C4The compound of: or its salt substituted to form the formula C5The compound of: or its salt, (e) general formula C5The nucleic acid or analog thereof is deprotected to form a nucleic acid-ligand conjugate of formula C6 or an analog thereof. In step (b) above, the equation C2PG ¹and PG ²The groups together with their intervening atoms form cyclic diol protecting groups, such as cyclic acetals or ketals. Such groups include methylene, ethylene, benzylidene, isopropylene, cyclohexylene, and cyclopentylene, silicone derivatives such as di-tertiary butylsilicon, and 1,1 , 3,3-tetraisopropyldisiloxane subunit (1,1,3,3-tetraisopropylidisiloxanylidene), cyclic carbonate, cyclic borate, and cyclic adenosine monophosphate (i.e. , cAMP) cyclic monophosphate derivative. In certain embodiments, the cyclic diol protecting group is 1,1,3,3-tetraisopropyldisiloxane subunit, which under basic conditions is from the formula C1Prepared by the reaction of diol and 1,3-dichloro-1,1,3,3-tetraisopropyldisiloxane. In step (c) above, the formula C2The nucleic acid or its analogue is alkylated with a mixture of DMSO and acetic anhydride under acidic conditions. In certain embodiments, when -V-H is a hydroxyl group, a mixture of DMSO and acetic anhydride undergoes Pumerel rearrangement in situ to form (methylthio)methyl acetate in the presence of acetic acid, and is then combined with formula C2hydroxyl reaction of nucleic acid or its analogues to provide the formula C3Monothioacetal functionalized fragment nucleic acid or analogs thereof. In step (d) above, use the formula C4Nucleic acid or its analog substitution formula C3The thiomethyl group of the nucleic acid or its analogues can be obtained by formula C4nucleic acid or its analogues. In certain embodiments, substitution occurs under mild oxidative and/or acidic conditions. In some embodiments, V is oxygen. In some embodiments, mild oxidizing agents include mixtures of elemental iodine and hydrogen peroxide, urea hydrogen peroxide complex, silver nitrate/silver sulfate, sodium bromate, ammonium peroxodisulfate, peroxodisulfate Tetrabutylammonium peroxydisulfate, Oxone®, Chloramine T, Selectfluor®, Selectfluor® II, sodium hypochlorite, or potassium iodate/sodium periodate. In certain embodiments, mild oxidizing reagents include N-iodosuccinimide, N-bromosuccinimide, N-chlorosuccinimide, 1,3-diiodo-5,5- Dimethylhydantion (1,3-diiodo-5,5-dimethylhydantion), pyridinium tribromide (pyridinium tribromide), iodine chloride or its complex, etc. Acids commonly used under mild oxidizing conditions include sulfuric acid, p-toluenesulfonic acid, trifluoromethanesulfonic acid, methanesulfonic acid, and trifluoroacetic acid. In certain embodiments, the mild oxidizing agent includes a mixture of N-iodosuccinimide and triflate. In step (e) above, remove the equation C5PG of nucleic acid-ligand conjugates or analogs thereof ³and R as necessary ⁴(when R ⁴When it is a suitable amine protecting group), we get the formula C6Nucleic acid-ligand conjugates or analogs or salts thereof. In some embodiments, PG ³and/or R ⁴Contains urethane derivatives, which can be removed under acidic or alkaline conditions. In certain embodiments, the formula C5Protecting groups for nucleic acid-ligand conjugates or analogs thereof (e.g., PG ³and R ⁴Both or independently PG ³or R ⁴either) are removed by acid hydrolysis. It should be understood that in the acid hydrolysis formula C5After the protecting group of the nucleic acid-ligand conjugate or its analogue, the formula is formed C6of salt. For example, when the formula is removed by treatment with an acid, such as hydrochloric acid C5When the acid-labile protecting group of the nucleic acid-ligand conjugate or its analog is used, the resulting amine compound will form its hydrochloride salt. One of ordinary skill in the art will recognize that a wide variety of acids can be used to remove acid-labile amine protecting groups, and therefore consider Eq. C6Various salt forms of nucleic acids or their analogs. In other specific embodiments, the formula C5Protective groups for nucleic acids or their analogs (e.g., PG ³and R ⁴Both or independently PG ³or R ⁴either) are removed by alkaline hydrolysis. For example, Fmoc and trifluoroacetyl protecting groups can be removed by treatment with a base. One of ordinary skill in the art will recognize that a wide variety of bases can be used to remove base-labile amine protecting groups. In some embodiments, the base is piperidine. In some specific embodiments, the base is 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). In certain embodiments, the formula C5The nucleic acid-ligand conjugate or its analog is deprotected under basic conditions and then treated with acid to form the formula C6of salt. In certain embodiments, the acid is butenedioic acid, formula C6The salt is fumarate. In some embodiments, the present disclosure provides methods for preparing oligonucleotide-ligand conjugates comprising one or more nucleic acid-ligand conjugates, or pharmaceutically acceptable salts thereof, the nucleic acid-ligand conjugates The conjugate unit is represented by formula II-b-3: This method includes the following steps: (a) Offering D5The oligonucleotide: or its salt, and, (b) Combine one or more adamantyl or lipophilic compounds with the formula D5oligonucleotides are combined to form a formula containing one or more nucleic acid-ligand conjugate units II-b-3oligonucleotide-ligand conjugates. In step (b) above, conjugation is carried out under suitable amide-forming conditions to obtain an adamantyl or lipid conjugate of the formula D5of compounds. Suitable amide forming conditions may include the use of amide coupling agents known in the art, such as, but not limited to, HATU, PyBOP, DCC, DIC, EDC, HBTU, HCTU, PyAOP, PyBrOP, BOP, BOP-Cl, DEPBT, T3P, TATU, TBTU, TNTU, TOTU, TPTU, TSTU, or TDBTU. In certain embodiments, amide forming conditions include HATU, and DIPEA or TEA. In some embodiments, the present disclosure provides for preparations comprising: D5Methods for oligonucleotide-ligand conjugates of the units represented: or its salt, the method includes the following steps: (a) Offering D4Nucleic acid-ligand conjugates or analogs thereof: or its salt, and (b) general formula D4The compound is deprotected to form the formula D5of compounds. In step (b) above, removing the PG of the oligonucleotide of formula D4 ³and R as necessary ⁴(when R ⁴When it is a suitable amine protecting group), we get the formula D5oligonucleotide-ligand conjugates or salts thereof. In some embodiments, PG ³and/or R ⁴Contains urethane derivatives which can be removed under acidic or alkaline conditions. In certain embodiments, the formula D4Protecting groups for oligonucleotide-ligand conjugates (e.g., PG ³and R ⁴Both or independently PG ³or R ⁴either) are removed by acid hydrolysis. It should be understood that in the acid hydrolysis formula D4After the protecting group of the oligonucleotide-ligand conjugate, its formula is formed D5of salt. For example, when the formula is removed by treatment with an acid, such as hydrochloric acid D4When the oligonucleotide has an acid-labile protecting group, the resulting amine compound will form its hydrochloride salt. One of ordinary skill in the art will recognize that a wide variety of acids can be used to remove acid-labile amine protecting groups, and therefore consider Eq. D5Various salt forms of nucleic acid-ligand conjugate units or analogs thereof. In other specific embodiments, the formula D4Protecting groups for oligonucleotide-ligand conjugates (e.g., PG ³and R ⁴Both or independently PG ³or R ⁴either) are removed by alkaline hydrolysis. For example, Fmoc and trifluoroacetyl protecting groups can be removed by treatment with a base. One of ordinary skill in the art will recognize that a wide variety of bases can be used to remove base-labile amine protecting groups. In some embodiments, the base is piperidine. In some specific embodiments, the base is 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). In some embodiments, the present disclosure provides for the preparation of oligonucleotide-ligand conjugates comprising one or more nucleic acid-ligand conjugate units having one or more adamantyl and/or lipid moieties or Its pharmaceutically acceptable salt method, the conjugate unit is represented by formula D4: This method includes the following steps: (a) Offering D3Nucleic acid or its analogues: or its salt, and (b) general formula D3The compound oligomericly polymerizes to form the formula D4of compounds. In the above step (b), oligopolymerization refers to using known and commonly used processes to perform oligopolymerization forming conditions to prepare oligonucleotides in the art. For example, consider the formula D3The nucleic acid or its analog is coupled to a solid support nucleic acid or its analog having a 5'-hydroxyl group. Further steps may include one or more deprotection, coupling, phosphite oxidation, and cleavage from the solid support to provide the adamantyl or lipid conjugate-containing formula of the present disclosure. D4The compounds represent oligonucleotides of various nucleotide lengths. In some embodiments, the present disclosure provides methods for preparing nucleic acids or analogs thereof comprising one or more lipid conjugates, further comprising preparing a formula D3Nucleic acid or its analogues: or its salt, the method includes the following steps: (a) Offering C5Nucleic acid or its analogues: or its salt, (b) general formula C5The nucleic acid or analog thereof is deprotected to form the formula D1The compound of: or its salt, (c) general formula D1The nucleic acid or its analog is protected to form the formula D2nucleic acid or its analogues. or its salt, and (d) general formula D2The nucleic acid or analog thereof is treated with a P(III) forming reagent to form the formula D3nucleic acid or its analogues. In step (b) above, the formula C5PG of nucleic acid or its analogues ¹and PG ²Contains silyl ethers or cyclic silene derivatives, which can be removed under acidic conditions or with fluoride anions. Examples of reagents that provide fluoride anions to remove silicon-based protecting groups include hydrofluoric acid, pyridine hydrogen fluoride, triethylamine trihydrofuride, tetrafluorofluoride N-Butylammonium, etc. In step (c) above, the formula D1The nucleic acid or analog thereof is protected with a suitable hydroxyl protecting group. In some embodiments, for protective D1The protective group PG of the 5'-hydroxyl group of the compound ⁴Includes acid-labile protecting groups such as trityl, 4-methoxytrityl, 4,4'-dimethoxytrityl, 4,4',4"-trimethoxytrityl base, 9-phenyl-𠮿-9-yl, 9-(p-phenylmethyl)-𠮿-9-yl, phenyl𠮿yl, 2,7-dimethylphenyl𠮿yl, etc. In some specific In embodiments, acid-labile protecting groups are suitable for deprotection using, for example, dichloroacetic acid or trichloroacetic acid during both solution and solid phase synthesis of acid-sensitive nucleic acids or analogs thereof. In the above step (d), the equation D2The nucleic acid or its analogue is treated with a P(III) forming reagent to obtain the formula D3of compounds. In the context of this disclosure, a P(III)-forming reagent is a phosphorus reagent that reacts to form a phosphorus(III) compound. In some embodiments, the P(III) forming reagent is 2-cyanoethyl N, N-Diisopropyl phosphoridite chloride or 2-cyanoethyl dichlorophosphate. In certain embodiments, the P(III) forming reagent is 2-cyanoethyl N, N-Diisopropylphosphoridite chloride. Those with ordinary knowledge should realize that in the form D2Compound X ¹Displacement of the leaving group in the P(III)-forming reagent is accomplished in the presence or absence of a suitable base. Such suitable bases are well known in the art and include organic and inorganic bases. In certain embodiments, the basic tertiary amine is such as triethylamine or diisopropylethylamine. In other embodiments, step (d) above uses N, N- Dimethylphosphoramidite dichloride is carried out as P(V) forming reagent. PreparationsVarious formulations have been developed to facilitate the use of oligonucleotides. For example, oligonucleotides (e.g., lipid-conjugated RNAi oligonucleotides) can be delivered to an individual or cellular environment using formulations that minimize degradation, facilitate delivery and/or uptake, or are The oligonucleotides provide another beneficial property. In some embodiments, provided herein are compositions comprising oligonucleotides (e.g., lipid-conjugated RNAi oligonucleotides) that reduce target mRNA (e.g., target mRNA expressed in stellate cells of the CNS). ) performance. Such compositions can be suitably formulated so that when administered to an individual (either into the immediate environment of a target cell or systemically), a sufficient portion of the oligonucleotide enters the cell to reduce target gene expression. Any of a variety of suitable oligonucleotide formulations may be used to deliver oligonucleotides to reduce stellate cell target gene expression as disclosed herein. In some embodiments, oligonucleotides are formulated in buffer solutions such as phosphate buffered saline solution, liposomes, microstructures, and shells. In some embodiments, the formulations herein include excipients. In some embodiments, excipients confer improved stability, improved absorption, improved solubility, and/or enhanced therapeutic properties of the active ingredient to the composition. In some embodiments, the excipient is a buffer (eg, sodium citrate, sodium phosphate, tris base, or sodium hydroxide) or vehicle (eg, buffered solution, paraffin oil, dimethyl sulfoxide, or mineral oil). In some embodiments, the oligonucleotide is freeze-dried to extend its storage life and then made into solution prior to use (eg, administration to an individual). Accordingly, an excipient in a composition comprising any of the oligonucleotides described herein can be a lyoprotectant (e.g., mannitol, lactose, polyethylene glycol, or polyvinylpyrrolidine ketones) or collapse temperature modifier (such as dextran, Ficoll™ or gelatin). Likewise, the oligonucleotides herein may be provided in their free acid form. In some embodiments, pharmaceutical compositions are formulated to be compatible with their intended route of administration. Examples of routes of administration include parenteral (e.g., intravenous, intramuscular, intraperitoneal, intradermal, subcutaneous, intrathecal), oral (e.g., inhalation), transdermal (e.g., topical), transmucosal, and rectal administration. give. In some embodiments, pharmaceutical compositions are formulated for delivery to the central nervous system (eg, intrathecal, epidural). In some embodiments, the pharmaceutical compositions are formulated for delivery to the eye (e.g., ophthalmic, intraocular, subconjunctival, intravitreal, retrobulbar, intracameral) ). Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (when water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.), or phosphate buffered saline (PBS). The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyols (eg, glycerol, propylene glycol, liquid polyethylene glycol, etc.), and suitable mixtures thereof. In many cases it will be preferable to include isotonic agents such as sugars, polyols (such as mannitol, sorbitol), sodium chloride in the composition. Sterile injectable solutions can be prepared by incorporating the oligonucleotide in the required amount and, if necessary, one or a combination of the ingredients enumerated above in a solvent of choice, followed by filtered sterilization. In some embodiments, the compositions may contain at least about 0.1% of a therapeutic agent (e.g., a lipid-conjugated RNAi oligonucleotide herein) or more, although the percentage of active ingredient(s) may be in the total. Between about 1% and about 80% or more of the weight or volume of the composition. Factors such as solubility, bioavailability, biological half-life, route of administration, product storage life, and other pharmacological considerations should be considered by one of ordinary skill in the art of preparing such pharmaceutical formulations, and thus may require various Dosage and treatment regimen. Instructions Reduce target gene expression In some specific embodiments, the present disclosure provides for contacting or delivering to a cell or population of cells an effective amount of any of the lipid-conjugated RNAi oligonucleotides herein to reduce star formation in the CNS. Methods for expression of target genes in cells. In some embodiments, expression of stellate cell target genes is reduced in regions of the CNS. In some embodiments, regions of the CNS include, but are not limited to, the brain, prefrontal cortex, frontal cortex, motor cortex, temporal cortex, parietal cortex, occipital cortex, sensory cortex, hippocampus, caudate Caudate, striatum, globus pallidus, optic thalamus, midbrain, tegmentum, substantia nigra, pons, brainstem, cerebellar white matter, cerebellum, Dentate nucleus, medulla oblongata, cervical spinal cord, thoracic spinal cord, lumbar spinal cord, cervical dorsal root ganglion, thoracic dorsal root ganglion, lumbar dorsal root ganglion, sacral dorsal root nerve Sacral dorsal root ganglion, nodose ganglia, femoral nerve, sciatic nerve, sural nerve, amygdala, hypothalamus, putamen, callosum, and cranial nerves. In some embodiments, the CNS region is selected from the group consisting of lumbar spinal cord, cerebellum, medulla oblongata, hippocampus, hypothalamus, frontal cortex, and combinations thereof. In some aspects, the region of the CNS is selected from the group consisting of spinal cord, lumbar spinal cord, thoracic spinal cord, cervical spinal cord, medulla oblongata, hippocampus, hypothalamus, cerebellum, frontal cortex, and combinations thereof. In some embodiments, lipid-conjugated RNAi oligonucleotides described herein reduce expression of target genes in stellate cells in the spinal cord. In some embodiments, lipid-conjugated RNAi oligonucleotides described herein reduce expression of target genes in stellate cells in the lumbar spinal cord. In some embodiments, lipid-conjugated RNAi oligonucleotides described herein reduce expression of target genes in stellate cells in the thoracic spinal cord. In some embodiments, lipid-conjugated RNAi oligonucleotides described herein reduce expression of target genes in cervical spinal cord stellate cells. In some embodiments, lipid-conjugated RNAi oligonucleotides described herein reduce expression of target genes in stellate cells in the hypothalamus. In some embodiments, lipid-conjugated RNAi oligonucleotides described herein reduce expression of target genes in stellate cells in the medulla oblongata. In some embodiments, lipid-conjugated RNAi oligonucleotides described herein reduce expression of target genes in stellate cells in the hippocampus. In some embodiments, lipid-conjugated RNAi oligonucleotides described herein reduce expression of target genes in stellate cells in the cerebellum. In some embodiments, lipid-conjugated RNAi oligonucleotides described herein reduce expression of target genes in stellate cells in the frontal cortex. In some embodiments, a decrease in target gene expression is determined by measuring a decrease in the amount or level of target mRNA, protein encoded by target mRNA, or target gene (mRNA or protein) activity in the cell. Such methods include those described herein and known to those of ordinary skill in the art. In some specific embodiments, the present disclosure provides for contacting or delivering an effective amount of any of the oligonucleotides herein (e.g., RNAi oligonucleotides) to a cell or population of cells to reduce GFAPMethod of performance. In some specific embodiments, GFAPReduction in performance was achieved by measuring the GFAPDetermined by a decrease in the amount or level of mRNA, GFAP protein, or GFAP activity. Such methods include those described herein and known to those of ordinary skill in the art. In some embodiments, the present disclosure provides for reducing stress in the central nervous system GFAPMethod of performance. In some embodiments, the central nervous system includes the brain and spinal cord. In some specific embodiments, GFAPManifestations are reduced in at least one area of the brain. In some embodiments, regions of the brain include the spinal cord, lumbar spinal cord, thoracic spinal cord, cervical spinal cord, medulla oblongata, cerebellum, hypothalamus, hippocampus, and frontal cortex. In some embodiments, a method for reducing expression of a target gene in stellate cells of the spinal cord of an individual comprises administering to the individual a lipid-conjugated RNAi oligonucleotide comprising a blunt end and at least one binding to the sense strand The lipid moiety at position 1, position 4, position 8, position 12, position 13, position 18, or position 20. In some embodiments, methods for reducing expression of a target gene in medulla oblongata stellate cells of an individual comprise administering to the individual a lipid-conjugated RNAi oligonucleotide comprising a blunt end and at least one binding to the sense strand The lipid moiety at position 4, position 8, position 12, position 13, position 18, or position 20. In some embodiments, methods for reducing expression of a target gene in cerebellar stellate cells of an individual comprise administering to the individual a lipid-conjugated RNAi oligonucleotide comprising a blunt end and at least one binding to the sense strand The lipid moiety at position 4, position 12, position 13, position 18, or position 20. In some embodiments, a method for reducing expression of a target gene in stellate cells of the inferior thalamus of an individual comprises administering to the individual a lipid-conjugated RNAi oligonucleotide comprising a blunt end and at least one binding site The lipid portion at position 1, position 4, position 12, position 13, position 18, or position 20 of the justice strand. In some embodiments, methods for reducing expression of a target gene in stellate cells of the frontal cortex of an individual comprise administering to the individual a lipid-conjugated RNAi oligonucleotide comprising a blunt end and at least one binding site The lipid part at position 4 of the justice strand. In some embodiments, methods for reducing expression of a target gene in stellate cells of the spinal cord of an individual comprise administering to the individual a lipid-conjugated RNAi oligonucleotide comprising a backbone-loop and at least one binding in The lipid portion of the justice strand at position 1, position 4, position 8, position 12, position 13, position 18, position 20, position 23, position 28, position 29, or position 30. In some embodiments, methods for reducing expression of a target gene in medulla oblongata stellate cells of an individual comprise administering to the individual a lipid-conjugated RNAi oligonucleotide comprising a backbone-loop and at least one binding in The lipid portion at position 1, position 4, position 18, position 20, position 23, position 28, position 29, or position 30 of the justice strand. The cerebellum of an individual comprises administering to the individual a lipid-bound RNAi oligonucleotide comprising a backbone-loop and at least one lipid bound at position 1, position 4, position 23, position 28, or position 29 of the sense strand part. In some embodiments, a method for reducing expression of a target gene in stellate cells of the inferior thalamus of an individual comprises administering to the individual a lipid-conjugated RNAi oligonucleotide comprising a blunt end and at least one binding site The lipid portion of the justice strand at position 1, position 1, position 4, position 12, position 13, position 18, position 20, position 23, position 28, position 29, or position 30. In some embodiments, methods for reducing expression of a target gene in stellate cells of the frontal cortex of an individual comprise administering to the individual a lipid-conjugated RNAi oligonucleotide comprising a blunt end and at least one binding site The lipid part at position 23 of the justice strand. In some embodiments, a single dose of an oligonucleotide or oligonucleotides described herein reduces the expression of stellate cell mRNA by at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 weeks. In some embodiments, a single dose of an oligonucleotide or pharmaceutical composition described herein reduces the expression of stellate cell mRNA for at least 4 weeks. In some embodiments, a single dose of an oligonucleotide or pharmaceutical composition described herein reduces the expression of stellate cell mRNA for at least 8 weeks. In some embodiments, a single dose of an oligonucleotide or pharmaceutical composition described herein reduces the expression of stellate cell mRNA for at least 12 weeks. In some embodiments, a single dose of an oligonucleotide or pharmaceutical composition described herein reduces the expression of stellate cell mRNA for at least 23 weeks. In some embodiments, a single dose of an oligonucleotide or pharmaceutical composition described herein reduces the expression of stellate cell mRNA for at least 26 weeks. In some embodiments, a single dose of an oligonucleotide or pharmaceutical composition described herein reduces the expression of stellate cell mRNA for at least 29 weeks. In some embodiments, a single dose of an oligonucleotide or pharmaceutical composition herein reduces the expression of stellate cell mRNA by 1 month, by 2 months, by 3 months, by 4 months, up to 5 months, up to 6 months, up to 7 months, up to 8 months, up to 9 months, up to 10 months, up to 11 months, up to 12 months months, up to 13 months, up to 14 months, up to 15 months, or up to 16 months. In some embodiments, a single dose of an oligonucleotide or pharmaceutical composition herein reduces the expression of stellate cell mRNA for up to 12 months. In some embodiments, a decrease in target gene expression is determined by measuring a decrease in the amount or level of target mRNA, protein encoded by target mRNA, or target gene (mRNA or protein) activity in the cell. Such methods include those described herein and known to those of ordinary skill in the art. The methods provided herein can be used with any appropriate cell type. In some embodiments, the cell line is any cell expressing a stellate cell target mRNA. In some embodiments, the cell line is obtained from an individual's primary stellate cell line. In some embodiments, the primary cells have undergone a limited number of passages such that the cells substantially maintain native phenotypic properties. In some embodiments, the cell line to which the oligonucleotide is delivered is ex vivo or ex vivo (ie, can be delivered to cells in culture or to the organism in which the cells reside). In some embodiments, lipid-conjugated RNAi oligonucleotides disclosed herein are delivered to cells or cell populations (e.g., stellate cells) using nucleic acid delivery methods known in the art including, but not limited to Injection of a solution or pharmaceutical composition containing lipid-conjugated RNAi oligonucleotides, exposure of cells or cell populations to particles containing lipid-conjugated RNAi oligonucleotides by bombardment, of RNAi oligonucleotide, or the cell membrane is electroporated in the presence of lipid-conjugated RNAi oligonucleotide. Other methods known in the art for delivering oligonucleotides to cells may be used, such as lipid-vectored vehicle delivery, chemical-vectored delivery, and cationic liposome transfection (such as calcium phosphate), among others. In some embodiments, target gene expression is reduced by an assay or technique that assesses one or more molecules, properties, or characteristics of a cell or cell population that correlates with target gene expression, or by assessing a direct indicator cell or cell population. An assay or technique is used to determine the molecules (e.g., target mRNA or protein) in which the target gene is expressed. In some embodiments, lipid-conjugated RNAi oligonucleotides provided herein reduce target gene expression in stellate cells to an extent by contacting the stellate cells with the lipid-conjugated RNAi oligonucleotide. cells or populations of stellate cells versus control cells or populations of cells (e.g., stellate cells or populations of stellate cells that are not contacted with a lipid-conjugated RNAi oligonucleotide or that are contacted with a control lipid-conjugated RNAi oligonucleotide) ) to evaluate by comparing the expression of target genes. In some embodiments, the control amount or level of target gene expression in a control cell or control cell population is predetermined such that the control amount or level does not need to be measured every time an assay or technique is performed. The predetermined level or value can take a variety of forms. In some embodiments, the predetermined level or value may be a single cutoff value, such as a median or mean. In some embodiments, contacting or delivering a lipid-conjugated RNAi oligonucleotide described herein to a stellate cell or population of stellate cells results in reduced expression of a stellate cell target gene. In some embodiments, the reduction in target gene expression is relative to a control of target gene expression in cells or cell populations that are not contacted with a lipid-conjugated RNAi oligonucleotide or that are contacted with a control lipid-conjugated RNAi oligonucleotide. quantity or level. In some specific embodiments, the target gene expression is reduced relative to the control amount or level of the target gene expression by about 1% or lower, about 5% or lower, about 10% or lower, about 15% or lower, About 20% or less, about 25% or less, about 30% or less, about 35% or less, about 40% or less, about 45% or less, about 50% or less, about 55% or less, about 60% or less, about 70% or less, about 80% or less, or about 90% or less. In some embodiments, the control amount or level of target gene expression is the amount or level of target mRNA and/or protein in cells or cell populations that have not been contacted with the lipid-conjugated RNAi oligonucleotides herein. In some embodiments, the effect of delivering a lipid-conjugated RNAi oligonucleotide to a cell or cell population according to the methods herein is over any limited period or amount of time (e.g., minutes, hours, days). , weeks, months). For example, in some embodiments, the target gene is expressed at least about 4 hours, about 8 hours, about 12 hours, about 18 hours after contacting or delivering the lipid-conjugated RNAi oligonucleotide to the cell or cell population. hours, about 24 hours; or at least about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 21 days, about 28 days, about 35 days, about 42 days, about 49 days, about 56 days, about 63 days, about 70 days, about 77 days, or about 84 days or more when measured in cells or cell populations. In some embodiments, the target gene is expressed at least about 1 month, about 2 months, about 3 months, about 3 months after contacting or delivering the lipid-conjugated RNAi oligonucleotide to the cell or cell population. Determined in cells or populations of cells at 4 months, about 5 months, or about 6 months or more. Treatment The present disclosure provides oligonucleotides for use as medicaments, and particularly for use in methods of treating diseases, disorders, and conditions associated with the CNS. The present disclosure also provides lipid-conjugated RNAi oligonucleotides for use in, or that may be applicable to, the treatment of individuals (e.g., humans) suffering from a disease, disorder or condition associated with expression of a stellate cell target gene. , disorder, or condition would benefit from reduced expression of stellate cell target genes. In some embodiments, the present disclosure provides lipid-conjugated RNAi oligonucleotides for use, or may be adapted to treat individuals suffering from a disease, disorder or condition associated with stellate cell target gene expression. The present disclosure also provides lipid-conjugated RNAi oligonucleotides for use in, or may be suitable for, the manufacture of medicaments or pharmaceutical compositions for the treatment of diseases, disorders or conditions associated with expression of stellate cell target genes. In some embodiments, lipid-conjugated RNAi oligonucleotides are used, or can be adapted to target mRNA and reduce expression of stellate cell target genes (eg, via the RNAi pathway). In some embodiments, lipid-conjugated RNAi oligonucleotides are used, or can be adapted to target mRNA and reduce the amount or level of stellate cell target mRNA, protein and/or activity. Furthermore, in some embodiments of the methods herein, individuals who have or are susceptible to a disease, disorder, or condition associated with expression of a stellate cell target gene are selected for use with the lipid-conjugated RNAi oligonucleotides herein. Glycoside treatment. In some embodiments, methods include selecting individuals who have or are susceptible to having markers (e.g., biomarkers) of a disease, disorder, or condition associated with expression of a stellate cell target gene, such as, but not limited to, the target mRNA, protein, or combinations thereof. Likewise, and as described in detail below, some embodiments of the methods provided by the present disclosure include steps such as measuring or obtaining baseline values for markers of expression of stellate cell target genes, and then comparing the values so obtained with a Or multiple other baseline values or values obtained after administration of a lipid-conjugated RNAi oligonucleotide to an individual are compared to assess the effectiveness of the treatment. The present disclosure also provides for the use of lipid-conjugated RNAi oligonucleotides provided herein to treat patients with, suspected of having, or developing a disease, disorder, or condition associated with expression of stellate cell target genes. Methods for expression of cellular target genes in individuals at risk for diseases, disorders, or conditions. In some specific embodiments, the present disclosure provides methods of using lipid-conjugated RNAi oligonucleotides provided herein to treat or attenuate the onset or progression of a disease, disorder, or condition associated with expression of a stellate cell target gene. In some embodiments, the present disclosure provides for the use of lipid-conjugated RNAi oligonucleotides provided herein to achieve a or A variety of therapeutic benefits. In some embodiments of the methods herein, an individual is treated by administering a therapeutically effective amount of any one or more of the lipid-conjugated RNAi oligonucleotides provided herein. In some embodiments, treatment includes reducing expression of stellate cell target genes (eg, in the CNS). In some embodiments, the subject is therapeutically treated. In some embodiments, the subject is treated prophylactically. In some embodiments of the methods herein, a lipid-conjugated RNAi oligonucleotide provided herein, or a pharmaceutical composition comprising a lipid-conjugated RNAi oligonucleotide, is administered to a patient with celiac disease. In an individual with a disease, disease or condition related to the expression of a target gene in a cell, the expression of the target gene in the individual is reduced, thereby treating the individual. In some embodiments, the amount or level of target mRNA is reduced in the individual. In some embodiments, the amount or level of protein encoded by the target mRNA is reduced in the individual. In some embodiments of the methods herein, a lipid-conjugated RNAi oligonucleotide provided herein, or a pharmaceutical composition comprising a lipid-conjugated RNAi oligonucleotide, is administered to a patient with celiac disease. In an individual with a disease, disorder or condition associated with expression of a cell target gene such that expression of the target gene in the individual is reduced compared to expression of the target gene before administration of a lipid-conjugated RNAi oligonucleotide or pharmaceutical composition At least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or greater than 99%. In some embodiments, when compared to individuals who did not receive a lipid-conjugated RNAi oligonucleotide or pharmaceutical composition or who received a control lipid-conjugated RNAi oligonucleotide, pharmaceutical composition, or treatment (e.g., reference or control individual), the expression of the stellate cell target gene in the individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, About 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or greater than 99%. In some embodiments of the methods herein, a lipid-conjugated RNAi oligonucleotide herein, or a pharmaceutical composition comprising a lipid-conjugated RNAi oligonucleotide, is administered to a patient with stellate cells. In an individual with a disease, disorder or condition associated with expression of the target gene, such that when compared to the amount or level of target mRNA before administration of the lipid-conjugated RNAi oligonucleotide or pharmaceutical composition, the amount of target mRNA in the individual is The amount or level is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, about 99%, or greater than 99%. In some embodiments, when compared to individuals who did not receive a lipid-conjugated RNAi oligonucleotide or pharmaceutical composition or who received a control lipid-conjugated RNAi oligonucleotide, pharmaceutical composition, or treatment (e.g., reference or a control individual), the amount or level of target mRNA in the individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60% , about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99%. In some embodiments of the methods herein, a lipid-conjugated RNAi oligonucleotide herein, or a pharmaceutical composition comprising a lipid-conjugated RNAi oligonucleotide, is administered to a patient with stellate cells. In an individual with a disease, disorder, or condition associated with expression of the target gene, such that when compared to the amount or level of the protein encoded by the target gene prior to administration of the lipid-conjugated RNAi oligonucleotide or pharmaceutical composition, the individual The amount or level of the protein encoded by the stellate cell target gene is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or greater than 99%. In some embodiments, when compared to individuals who did not receive a lipid-conjugated RNAi oligonucleotide or pharmaceutical composition or who received a control lipid-conjugated RNAi oligonucleotide, pharmaceutical composition, or treatment (e.g., ref. or the amount or level of the protein encoded by the target gene in the individual), the amount or level of the protein encoded by the stellate cell target gene in the individual is reduced by at least about 30%, about 35%, about 40%, about 45 %, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or greater than 99% . In some embodiments of the methods herein, a lipid-conjugated RNAi oligonucleotide herein, or a pharmaceutical composition comprising a lipid-conjugated RNAi oligonucleotide, is administered to a patient with stellate cells. In an individual with a disease, disorder, or condition associated with the expression of the target gene, such that the target gene activity in the individual is reduced when compared to the amount or level of target gene activity before administration of the lipid-conjugated RNAi oligonucleotide or pharmaceutical composition. The amount or level is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, About 85%, about 90%, about 95%, about 99%, or greater than 99%. In some embodiments, when compared to individuals who did not receive a lipid-conjugated RNAi oligonucleotide or pharmaceutical composition or who received a control lipid-conjugated RNAi oligonucleotide, pharmaceutical composition, or treatment (e.g., reference or a control individual), the amount or level of target gene activity in the individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or greater than 99%. Suitable methods for determining the expression of a target gene, the amount or level of target mRNA, the amount or level of protein encoded by the target gene, and/or the amount or level of target gene activity in an individual or in a sample from an individual are within the art. known in the field. Furthermore, the examples shown herein illustrate exemplary methods for determining expression of a gene of interest. In some embodiments, target gene expression, the amount or level of target gene mRNA, the amount or level of protein encoded by the target gene, the amount or level of target gene activity, or any combination thereof, is obtained or isolated from an individual. Cells (e.g., stellate cells), cell populations or groups of cells (e.g., organoids), organs (e.g., CNS), blood or portions thereof (e.g., plasma), tissues (e.g., brain tissue), samples (e.g., brain tissue) For example, CSF samples or brain biopsy samples), or any other biological material. In some embodiments, the expression of the stellate cell target gene, the amount or level of the target gene mRNA, the amount or level of the protein encoded by the target gene, the amount or level of the target gene activity, or any combination thereof, is different from that in the individual. More than one type of cell (e.g., stellate cells), more than one population of cells, more than one organ (e.g., brain and one or more other organs), more than one blood fraction (e.g., plasma and one or more other organs) obtained or isolated or multiple other blood parts), more than one type of tissue (e.g., brain tissue and one or more other types of tissue), more than one type of sample (e.g., brain biopsy sample and one or more other types of biologic sample) Medium to low. In some embodiments, expression of the stellate cell target gene is reduced in one or more of the spinal cord, lumbar spinal cord, thoracic spinal cord, cervical spinal cord, medulla oblongata, hippocampus, hypothalamus, sensory cortex, or cerebellum. In some embodiments, expression of stellate cell target genes is reduced in the spinal cord. In some embodiments, expression of stellate cell target genes is reduced in the lumbar spinal cord. In some embodiments, expression of stellate cell target genes is reduced in the thoracic spinal cord. In some embodiments, expression of stellate cell target genes is reduced in the cervical spinal cord. In some embodiments, expression of stellate cell target genes is reduced in the hypothalamus. In some embodiments, expression of stellate cell target genes is reduced in the medulla oblongata. In some embodiments, expression of stellate cell target genes is reduced in the hippocampus. In some embodiments, expression of stellate cell target genes is reduced in sensory cortex. In some embodiments, expression of stellate cell target genes is reduced in the cerebellum. In some embodiments, expression of stellate cell target genes is reduced in the frontal cortex. Examples of diseases, disorders, or conditions associated with expression of stellate cell target genes include, but are not limited to, Spinocerebellar Atxia 1, Spinocerebellar Atxia 2, Spinocerebellar Atxia 3, Universal Prion Disease, Alexander's Disease, MECP2 Duplication Syndrome, Huntington's Disease, Parkinson's Disease, Alexander's Disease, Progress Progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), argyrophilic grain disease (AGD), global glial tauopathy, GGT), aging-related tau astrogliopathy (ARTAG), Familial Frontotemporal Dementia 17 (FTD-17), neurodegenerative diseases with respiratory failure (Tauopathy with Respiratory Failure), Dementia with Seizure, Pick's disease, Myotonic dystrophy 1 or 2, MD1 or MD2 ), Down's syndrome, spastic paraplegia (SP), Niemann-Pick disease type C, Dementia with Lewy bodies (DLB), Lewy body dysphagia, Lewy body disease, Olivopontocerebellar atrophy, Sriatonigral degeneration, Schail-De Shy-Drager syndrome, Spinal muscular atrophy V (SMAV), Huntington's disease (HD), Alzheimer's Disease, SCA1, SCA2, SCA3, SCA7, SCA10 (spinocerebellar ataxia type 1, 2, 3, 7 or 10), multiple system atrophy (MSA), spinal cord Spinal and Bulbar Muscular Atrophy (SBMA, Kennedy disease), Friedrich Ataxia, Fragile X-associated tremor /ataxia syndrome (FXTAS), Fragile X syndrome (FRAXA), X-Linked Mental Retardation (XLMR), Parkinson's Disease, dystonia, SBMA (spinal cord Spinobulbar muscular atrophy, neuropathic pain disorder, spinal cord injury, Dentatorubral-pallidoluysian atrophy (DRPLA), latent CNS disorders (recessive CNS disorder), ALS (amyotrophic lateral sclerosis), M2DS (MECP2 duplication syndrome), FTD (frontotemporal dementia), general Prion disease, Adult Onset Leukodystrophy, Alexander's Disease, Krabbe Disease, Chronic Traumatic Encephalopathy, Familial Pelizaeus-Merzbacher disease (PMD), Lafora disease, stroke, Cerebral Amyloid Angiopathy (CAA), and Metachromatic Leukodystrophy (MLD) ). Examples of diseases, disorders, or conditions associated with expression of stellate cell target genes include, but are not limited to, spinocerebellar disorders 1, spinocerebellar disorders 2, spinocerebellar disorders 3, Prion disease, Alexander's disease disease, MECP2 duplication syndrome, Huntington's disease, Parkinson's disease, and Alzheimer's disease. In some embodiments, the stellate cell target gene may be a target gene from any mammal, such as a human. Any stellate cell gene can be silenced according to the methods described herein. The methods described herein generally involve administering to an individual a therapeutically effective amount of a lipid-conjugated RNAi oligonucleotide herein, that is, an amount capable of producing the desired therapeutic outcome. A therapeutically acceptable amount may be an amount that therapeutically treats a disease or condition. The appropriate dosage for any individual will depend on factors including the individual's size, body surface area, age, the composition to be administered, the active ingredient(s) in the composition, the time of administration, and route, general health status, and other medications to be administered concurrently. In some embodiments, the subject is administered any of the compositions herein: enterally (e.g., orally, by a gastric feeding tube, by a duodenal feeding tube) tube), via gastrostomy, or transrectally), parenterally (e.g., subcutaneous injection, intravenous injection or infusion, intraarterial injection or infusion, intraosseous infusion, intramuscular injection, intracranial injection, intracerebroventricular injection , intrathecally), topically (e.g., on the skin, inhaled, via eye drops, or through mucous membranes), or by direct injection into the target organ (e.g., the brain of an individual). In some embodiments, the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, are administered intrathecally into the cerebrospinal fluid (CSF) (injection or infusion into body fluids within the subarachnoid space). )middle. In some embodiments, intrathecal administration of the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, is performed as a single bolus injection into the subarachnoid space. In some embodiments, intrathecal administration of the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, is performed by infusion into the subarachnoid space. In some embodiments, intrathecal administration of the present invention, or compositions thereof, is performed via a catheter into the subarachnoid space. In some embodiments, intrathecal administration of the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, is via a pump. In some embodiments, intrathecal administration of the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, is via an implantable pump. In some embodiments, administration is via an implantable device that operates or functions as a reservoir. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, are administered intrathecally into the cisterna magna (also known as the cisterna magna). Intrathecal administration into the cisterna magna is called "intracisternal administration" or "intracisternal magna administration". In some embodiments, the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, are administered intrathecally into the subarachnoid space of the lumbar spinal cord. Intrathecal administration into the subarachnoid space of the lumbar spinal cord is called "lumbar intrathecal (i.t.) administration". In some embodiments, the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, are administered intrathecally into the subarachnoid space of the cervical spinal cord. Intrathecal administration into the subarachnoid space of the cervical spinal cord is called "cervical intrathecal (i.t.) administration". In some embodiments, the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, are administered intrathecally into the subarachnoid space of the thoracic spinal cord. Intrathecal administration into the subarachnoid space of the thoracic spinal cord is called "thoracic intrathecal (i.t.) administration". In some embodiments, the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, are administered by intracerebroventricular injection or infusion into the cerebral ventricles. Intracerebroventricular administration into the ventricular space is called "intracerebroventricular (i.c.v.) administration". In some embodiments, Ommaya depots are used to administer the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, by intracerebroventricular injection or infusion. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, are administered annually, every 6 months, every 4 months, or quarterly. Once (every three months), every two months (every 2 months), monthly, or weekly. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, are administered weekly or at intervals of two or three weeks. In some embodiments, the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, are administered daily. In some embodiments, the subject is administered one or more loading doses of a lipid-conjugated RNAi oligonucleotide, or a composition thereof, followed by one of the lipid-conjugated RNAi oligonucleotides or multiple maintenance doses. In some embodiments, the subject to be treated is a human, or non-human primate, or other mammalian subject. Other exemplary individuals include domestic animals, such as dogs and cats; domestic animals, such as horses, cattle, pigs, sheep, goats, and chickens; and animals, such as mice, rats, guinea pigs, and hamsters.setIn some embodiments, the present disclosure provides kits comprising lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, and instructions for use. In some embodiments, a kit includes a lipid-conjugated RNAi oligonucleotide herein, or a composition thereof, and a package insert containing instructions for use of the kit and/or any component thereof. In some embodiments, a kit includes a lipid-conjugated RNAi oligonucleotide herein, or compositions thereof, one or more controls, and various buffers, reagents, enzymes, and Other standard ingredients well known in the art. In some embodiments, the container includes at least one vial, well, test tube, flask, bottle, syringe, or other container device into which a lipid-conjugated RNAi oligonucleotide, or composition thereof, herein is placed, and in some cases by appropriate aliquots. In some embodiments where additional components are provided, the kit contains additional containers in which the components are placed. Kits may also include devices strictly restricted for commercial sale containing the lipid-conjugated RNAi oligonucleotides herein, or compositions thereof, and any other reagents. Such containers may include injection molded or blow molded plastic containers in which the desired vial is retained. Containers and/or kits may include labels with instructions for use and/or warnings. In some embodiments, a kit includes a lipid-conjugated RNAi oligonucleotide herein, or a composition thereof, and a pharmaceutically acceptable carrier, or a pharmaceutical containing a lipid-conjugated RNAi oligonucleotide. Compositions and instructions for treating or delaying the progression of a disease, disorder or condition associated with expression of a stellate cell target gene in an individual in need thereof. [ definition ]As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items. In addition, the singular form and the articles "a/an/" and "the" are intended to include the plural form as well, unless expressly stated otherwise. It should further be understood that the terms include, comprise, including and/or comprising, when used in this specification, indicate the presence of stated features, integers, steps, operations, elements, and /or components, but not excluding the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof. Furthermore, it will be understood that when an element is referred to as a component or subsystem and/or shown as being connected or coupled to another element, it can be directly connected or coupled to the other element or intervening elements may be present. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods or compositions, the methods and materials described herein are exemplary. General textbooks describing molecular biology techniques useful in this article, including the use of vectors, promoters, and many other related topics, include Berger and Kimmel, GUIDE TO MOLECULAR CLONING TECHNIQUES, Methods in Enzymology, volume 152, (Academic Press, Inc. , San Diego, Calif.) ("Berger"); Sambrook et al., Molecular Cloning--A Laboratory Manual ,2d ed., Vol. 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor, 1989 ("Sambrook") and Current Protocols in Molecular Biology, F.M.Ausubel et al., eds., CURRENT PROTOCOLS, A JOINT VENTURE BETWEEN GREENE PUBLISHING ASSOCIATES, INC. AND JOHN WILEY AND SONS, INC.,(1999 Supplement) ("Ausubel"). Sufficient to guide those skilled in the art through in vitro amplification methods (including polymerase chain reaction (PCR), ligase chain reaction (LCR), Q.beta replicase amplification and other RNA polymerase media Examples of techniques (e.g., NASBA) such as procedures for producing homologous nucleic acids of the present disclosure are found in: Berger, Sambrook, and Ausubel, and in Mullis et al., (1987) U.S. Patent No. 4,683,202; Innis et al., eds. (1990); PCR PROTOCOLS: A GUIDE TO METHODS AND APPLICATIONS (Academic Press Inc. San Diego, Calif.) ("Innis"); Arnheim and Levinson (Oct. 1, 1990) CandEN 36-47; J. NIH RES.(1991)3:81-94; Kwoh et al., (1989) PROC.NATL.ACAD.SCI.USA 86: 1173; Guatelliet al(1990) PROC.NAT'L.ACAD.SCI.USA 87: 1874; Lomell et al., (1989) J. CLIN.CHEM 35: 1826; Landegren et al., (1988) SCIENCE 241: 1077-80; Van Brunt (1990) BIOTECHNOLOGY 8: 291-94; Wu and Wallace (1989) GENE 4: 560; Barringer et al., (1990) GENE 89:117; and Sooknanan and Malek (1995) BIOTECHNOLOGY 13: 563-564. inWallace et al., US Patent No. 5,426,039 describes an improved method for amplifying nucleic acids in vitro. In Cheng et al., (1994) NATURE 369: 684-85 and the literature cited therein summarize improved methods for the amplification of large nucleic acids by PCR, in which PCR amplicons of up to 40 kb are generated. As used in this specification and the accompanying claims, the singular forms "a/an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a pharmaceutical carrier" includes a mixture of two or more such carriers and the like. Ranges may be expressed herein as from "about" one value, and/or to "about" another value. When expressed as such a range, another embodiment includes from one value and/or to the other value. Similarly, when an approximation of a numerical value is expressed by use of the preceding "about", it should be understood that the value forms another embodiment. It is further understood that the endpoints of each range are meaningful whether viewed in relation to the other endpoint or independently of the other endpoint. It is also understood that every value disclosed herein and in addition to the value itself, each value disclosed herein is also "about" that value. For example, if "10" is revealed, "approximately 10" is also revealed. It is also to be understood that when a value is disclosed, "less than or equal to" the value, "greater than or equal to" the value, and possible ranges between values are also disclosed, as would be understood by one of ordinary skill in the art. . For example, if the value "10" is revealed, then "less than or equal to 10" and "greater than or equal to 10" are also revealed. Also understand that throughout the application, data is provided in several different formats, and that this data represents endpoints and starting points, as well as ranges for any combination of those data points. For example, if a particular data point of "10" is disclosed and a particular data point of "15" is disclosed, it will be understood that the disclosure is considered to be greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15, and between 10 and 15. Also understand that units between two specific units are also revealed. For example, if 10 and 15 are disclosed, 11, 12, 13, and 14 are also disclosed. As used herein, "complementary" refers to a relationship between two nucleotides (e.g., on two opposing nucleic acids or on opposing regions of a single nucleic acid strand) that allows the two nucleotides to form a base with each other. Structural relationship of base pairs. For example, purine nucleotides of one nucleic acid that are complementary to pyrimidine nucleotides of an opposing nucleic acid can base pair together by forming hydrogen bonds with each other. In some embodiments, complementary nucleotides may be base paired in a Watson-Click manner or in any other manner that allows the formation of stable duplexes. In some embodiments, two nucleic acids can have regions of multiple nucleotides that are complementary to each other to form a region of complementarity, as described herein. As used herein, "deoxyribonucleotide" refers to a nucleotide that has a hydrogen in place of a hydroxyl group at the 2' position of its pentose sugar when compared to ribonucleotides. Modified deoxyribonucleotides are deoxyribonucleotides having modifications or substitutions of one or more atoms other than at the 2' position, including modifications within or on sugars, phosphate groups or bases or replace. As used herein, "double-stranded RNA/dsRNA/dsRNAi" refers to an RNA oligonucleotide that is substantially in the form of a duplex. In some embodiments, complementary base pairing of the double-stranded region(s) of a dsRNA oligonucleotide is formed between antiparallel sequences of nucleotides in covalently separated nucleic acid strands. In some embodiments, complementary base pairing of the double-stranded region(s) of the dsRNA is formed between antiparallel sequences of nucleotides of the covalently linked nucleic acid strands. In some embodiments, the complementary base pairing of the double-stranded region(s) of the dsRNA is formed from a single nucleic acid strand that is folded (e.g., via a hairpin) to provide nucleotides that are base paired together. of complementary antiparallel sequences. In some embodiments, the dsRNA comprises two covalently separated nucleic acid strands that are fully double-stranded from each other. However, in some embodiments, the dsRNA comprises two covalently separated nucleic acid strands that are partially double-stranded (eg, have overhangs at one or both ends). In some embodiments, the dsRNA contains antiparallel sequences of partially complementary nucleotides and, therefore, may have one or more mismatches, which may include internal mismatches or terminal mismatches. As used herein, a "duplex" with respect to a nucleic acid (eg, an oligonucleotide) refers to a structure formed by complementary base pairing of two antiparallel sequences of nucleotides. As used herein, "excipient" refers to a non-therapeutic agent that may be included in a composition, for example, to provide or contribute to a desired consistency or stabilizing effect. As used herein, a "loop" refers to an unpaired region of a nucleic acid (e.g., an oligonucleotide) that is flanked by antiparallel regions of two nucleic acids that are antiparallel to each other. The parallel regions are sufficiently complementary to each other that under appropriate hybridization conditions (e.g., in phosphate buffer solution, in cells), two antiparallel regions flanking the unpaired region hybridize to form a duplex (called a duplex). "trunk"). As used herein, "GFAP" refers to glial fibrillary acidic protein (Glial Fibrillary Acidic Protein). GFAP is found in stellate cells of the central nervous system (CNS). GFAP is an intermediate filament protein that provides support for cell structure. As used herein, "reduction of GFAP expression" refers to the amount or level of GFAP mRNA, GFAP protein, and/or GFAP activity in a cell, cell population, sample, or individual when compared to an appropriate Reduced by reference (eg, reference cell, cell population, sample, or individual). As used herein, "modified internucleotide linkage" refers to one or more chemical modifications when compared to a reference internucleotide linkage that includes a phosphodiester bond linkages between nucleotides. In some embodiments, the modified nucleotides are non-naturally occurring linkages. Generally speaking, a modified internucleotide linkage confers one or more desirable properties to the nucleic acid in which the modified internucleotide linkage is present. For example, modified nucleotides can improve thermal stability, resistance to degradation, nuclease resistance, solubility, bioavailability, biological activity, reduced immunogenicity, etc. As used herein, "modified nucleotide" refers to a nucleotide that has one or more chemical modifications when compared to a corresponding reference nucleotide selected from: adenine ribose Nucleotides, guanine ribonucleotides, cytosine ribonucleotides, uracil ribonucleotides, adenine deoxyribonucleotides, guanine deoxyribonucleotides, cytosine deoxyribonucleotides acid, and thymidine deoxyribonucleotides. In some embodiments, the modified nucleotides are non-naturally occurring nucleotides. In some embodiments, modified nucleotides have one or more chemical modifications in their sugar, nucleobase, and/or phosphate groups. In some embodiments, modified nucleotides have one or more chemical moieties that bind to corresponding reference nucleotides. Generally speaking, modified nucleotides confer one or more desired properties on the nucleic acid in which the modified nucleotide is present. For example, modified nucleotides can improve thermal stability, resistance to degradation, nuclease resistance, solubility, bioavailability, biological activity, reduced immunogenicity, etc. As used herein, "astrocyte mRNA" and "astrocyte gene" refer to any gene, mRNA, or gene encoded/expressed by genes in stellate cells of the central nervous system. and/or protein. Stellate cells are glial cells that perform several functions throughout the CND, such as synaptic support and neurotransmission. As used herein, "nicked tetraloop structure" refers to an RNAi oligonucleotide structure characterized by separate sense (follower) strands and antisense (leader) strands, where the sense strand has Regions that are complementary to the antisense strands and in which at least one of the strands (usually the sense strand) has four loops configured to stabilize the adjacent backbone region formed within the at least one strand. As used herein, "oligonucleotide" refers to a short nucleic acid (eg, less than about 100 nucleotides long). Oligonucleotides can be single-stranded (ss) or ds. Oligonucleotides may or may not have double-stranded regions. As a non-limiting set of examples, oligonucleotides may be, but are not limited to, small interfering RNA (siRNA), microRNA (miRNA), short hairpin RNA (shRNA), Dicer matrix interfering RNA (dsiRNA), antisense Oligonucleotides, short siRNA or ss siRNA. In some embodiments, the double-stranded (dsRNA) is an RNAi oligonucleotide. The terms "lipid-conjugated RNAi oligonucleotide" and "oligonucleotide-ligand conjugate" are used interchangeably and refer to a combination of one or more Or oligonucleotides that target ligand (eg, lipid) binding nucleotides. As used herein, an "overhang" refers to the terminal non-base-paired nucleotide(s) resulting from a strand or region extending beyond the terminus of the complementary strand that is consistent with the complementary strand The ends form double strands. In some embodiments, the overhang comprises one or more unpaired nucleotides extending from the double-stranded region at the 5' end or the 3' end of the dsRNA. In some embodiments, the overhang is a 3' or 5' overhang on the antisense or sense strand of the dsRNA. As used herein, "phosphate analog" refers to a chemical moiety that mimics the electrostatic and/or steric properties of a phosphate group. In some embodiments, the phosphate analog is positioned at the 5' terminal nucleotide of the oligonucleotide, replacing the 5'-phosphate that is normally susceptible to enzymatic removal. In some embodiments, the 5' phosphate analog contains a phosphatase-resistant linkage. Examples of phosphate analogs include, but are not limited to, 5'phosphonate esters such as 5'methylenephosphonate (5'-MP) and 5'-(E)-vinylphosphonate (5'-VP ). In some embodiments, the oligonucleotide has a phosphate analog at the 4'-carbon position of the sugar at the 5' terminal nucleotide (termed a "4'-phosphate analog"). An example of a 4'-phosphate analog is an oxymethylphosphonate in which the oxygen atom of the oxymethyl group is bonded to a sugar moiety (eg, at its 4'-carbon) or analog thereof. See, for example, U.S. Provisional Patent Application Nos. 62/383,207 (filed on September 2, 2016) and 62/393,401 (filed on September 12, 2016). Other modifications targeting the 5' end of oligonucleotides have been developed (see, e.g., International Patent Application No. WO 2011/133871; U.S. Patent No. 8,927,513; and Prakash et al.,(2015)NUCLEIC ACIDS RES .43:2993-3011). As used herein, "reduced expression" of a target gene refers to a cell, cell population, sample, or individual when compared to an appropriate reference (e.g., a reference cell, cell population, sample, or individual) The amount or level of RNA transcript (e.g., target mRNA) or protein encoded by the target gene is reduced and/or the amount or level of gene activity is reduced. For example, when compared to cells that were not treated with a double-stranded oligonucleotide, an oligonucleotide or conjugate herein (e.g., a lipid-conjugated RNAi oligonucleotide that contains a target mRNA that contains The action of contacting a cell with an antisense strand of a nucleotide sequence that is complementary to the nucleotide sequence can result in the target mRNA, the protein encoded by the target gene, and/or the activity of the target gene (e.g., inactivation of the target mRNA via the RNAi pathway and/or degradation). Similarly, and as used herein, "reducing expression" refers to behavior that results in reduced expression of a target gene. As used herein, a "region of complementarity" refers to a nucleotide sequence of a nucleic acid (e.g., dsRNA) that is sufficiently complementary to an antiparallel sequence of nucleotides to permit hybridization under appropriate hybridization conditions (e.g., , in phosphate buffer, in cells, etc.) to hybridize between two nucleotide sequences. In some embodiments, oligonucleotides herein comprise a targeting sequence having a region complementary to an mRNA target sequence. As used herein, "ribonucleotide" refers to a nucleotide having ribose as its pentose sugar, containing a hydroxyl group at its 2' position. Modified ribonucleotides are ribonucleotides that have modifications or substitutions of one or more atoms other than at the 2' position, including modifications or substitutions within or themselves of the ribose, phosphate group, or base. As used herein, "RNAi oligonucleotide" refers to (a) a dsRNA having a sense strand (follower) and an antisense strand (leader), in which the antisense strand or a portion of the antisense strand is derived from Used to cleave target mRNA by Ago2 endonuclease, or (b) ss oligonucleotide with a single antisense strand, wherein the antisense strand (or part of the antisense strand) is generated by Ago2 endonuclease is used to cleave target mRNA. As used herein, "strand" refers to a single, contiguous sequence of nucleotides linked together by internucleotide linkages (eg, phosphodiester linkages or phosphorothioate linkages). In some embodiments, the strand has two free ends (eg, a 5' end and a 3' end). As used herein, "subject" means any mammal, including mice, rabbits, and humans. In a specific embodiment, the subject is human or NHP. In addition, "individual" or "patient" may be used interchangeably with "subject". As used herein, "synthetic" means artificially synthesized (e.g., using machines (e.g., solid-state nucleic acid synthesizers)) or otherwise not derived from natural sources (e.g., cells or organisms) from which molecules are typically produced. Nucleic acids or other molecules. As used herein, "targeting ligand" refers to a molecule that selectively binds to a homologous molecule (e.g., a receptor) in a tissue or cell of interest and/or can bind to another substance to achieve A molecule or "moiety" (eg, carbohydrate, amino sugar, cholesterol, polypeptide, or lipid) that targets another substance to a tissue or cell of interest. For example, in some embodiments, a targeting ligand can be combined with an oligonucleotide for the purpose of targeting the oligonucleotide to a specific tissue or cell of interest. In some embodiments, the targeting ligand selectively engages a cell surface receptor. Accordingly, in some embodiments, the targeting ligand, when bound to the oligonucleotide, does so by selectively engaging a receptor expressed on the cell surface and consisting of the oligonucleotide, the targeting ligand, and the receptor. The cells of the somatic complex undergo endosomal internalization to facilitate delivery of oligonucleotides to specific cells. In some embodiments, the targeting ligand binds to the oligonucleotide via a linker that is cleaved after or during cellular internalization, such that the oligonucleotide releases the targeting ligand in the cell. As used herein, "tetraloop" refers to a loop that increases the stability of adjacent duplexes formed by hybridization of flanking sequences of nucleotides. The stability can be increased by increasing the melting temperature (T) of adjacent backbone duplexes. _m), that is, the melting temperature ratio derived from the average T of adjacent backbone duplexes expected from a set of loops of comparable length consisting of randomly selected nucleotide sequences _mhigh. For example, tetracycline can be dissolved in 10 mM NaHPO ₄At least about 50°C, at least about 55°C, at least about 56°C, at least about 58°C, at least about 60°C, at least about 65°C, or at least about 75°C T _m. In some embodiments, tetracycles can stabilize bp in adjacent backbone duplexes through stacking interactions. In addition, interactions between nucleotides in the four-ring ring include, but are not limited to, non-Watson-Crick base pairing, stacking interactions, hydrogen bonding and contact interactions (Cheong et al.,(1990)NATURE 346:680-82; Heus and Pardi (1991) SCIENCE 253:191-94). In some embodiments, the tetracycle contains or consists of 3 to 6 nucleotides, and typically 4 to 5 nucleotides. In some embodiments, a tetracycle includes or consists of 3, 4, 5 to 6 nucleotides, which may or may not be modified (e.g., which may or may not be associated with the target partially combined). In a specific embodiment, the tetracycle consists of 4 nucleotides. Any nucleotide can be used in the tetracycle and can be used as in Cornish-Bowden ((1985) NUCLEIC ACIDS RES .The standard IUPAC-IUB notation for such nucleotides is used as described in 13:3021-3030). For example, the letter "N" can be used to mean that any base can be at that position, the letter "R" can be used to show that A (adenine) or G (guanine) can be at that position, and "B" can be used to It shows that C (cytosine), G (guanine), and T (thymine) can be located at this position. Examples of four-rings include the UNCG family of four-rings (e.g., UUCG), the GNRA family of four-rings (e.g., GAAA), and the CUUG four-rings (Woese et al.,(1990) PROC.NATL. ACAD.SCI.USA 87:8467-71; Antao et al.,(1991) NUCLEIC ACIDS RES .19:5901-05). Examples of DNA tetracycles include the d(GNNA) family of tetracycles (e.g., d(GTTA), d(GNRA) tetracycles) family, the d(GNAB) family of tetracycles, and the d(CNNG) family of tetracycles , and the d(TNCG) family of four rings (for example, d(TTCG)). (See e.g., Nakano et al.,(2002)BIOCHEM. 41:4281-92; Shinji et al.,(2000) NIPPON KAGAKKAI KOEN YOKOSHU 78:731). In some embodiments, the four-ring system is contained within a gapped four-ring structure. As used herein, "treating" means providing treatment to an individual for the purpose of improving the health and/or well-being of an individual with an existing condition (e.g., disease, disorder) or preventing or reducing the likelihood of the condition occurring. The act of providing care to an individual in need thereof, such as by administering a therapeutic agent (eg, an oligonucleotide herein) to the individual. In some embodiments, treatment involves reducing the frequency or severity of at least one sign, symptom, or contributing factor of a condition (eg, disease, disorder) experienced by an individual. Example Example 1 : RNAi Preparation of oligonucleotides Oligonucleotide synthesis and purificationThe oligonucleotides (RNAi oligonucleotides) described in the preceding examples were chemically synthesized using methods described herein. In principle, RNAi oligonucleotides are synthesized using solid-phase oligonucleotide synthesis methods as described for 19 to 23mer RNAi oligonucleotides (see, e.g., Scaringe et al.(1990) NUCLEIC ACIDS RES .18:5433-41 and Usman et al.(1987)J. AM. CHEM.SOC .109:7845-45, see also, U.S. Patent Nos. 5,804,683; 5,831,071; 5,998,203; 6,008,400; 6,111,086; 6,117,657; 6,353,098; 6,362,323; 6,437,117 No. 6,469,158) , and using known phosphoramidite synthesis methods (see, e.g., Hughes and Ellington (2017) COLD SPRING HARB PERSPECT BIOL .9(1):a023812; Beaucage S.L., Caruthers M.H. STUDIES ON NUCLEOTIDE CHEMISTRY V : Deoxynucleoside Phosphoramidites-A New Class of Key I ntermediates for Deoxypolynucleotide Synthesis, TETRAHEDRON LETT. 1981;22:1859-62. doi: 10.1016/S0040-4039(01) 90461-7); PCT Application No. PCT/US2021/42469 (each of which is incorporated herein by reference). RNAi oligonucleotides with a 19mer core sequence are arranged to have a 36mer sense strand and a 22mer antisense strand for processing by the RNAi machinery. The 19mer core sequence is complementary to a region in the GFAP mRNA. Individual RNA strands were synthesized and subjected to HPLC purification according to standard methods (Integrated DNA Technologies; Coralville, IA). For example, RNA oligonucleotides were synthesized using solid phase phosphoramidite chemistry, deprotection, and deprotection on a NAP-5 column (Amersham Pharmacia Biotech; Piscataway, NJ) using standard methods. Salt (Damha & Olgivie (1993) METHODS MOL. BIOL. 20:81-114; Wincott et al.(1995) NUCLEIC ACIDS RES.23:2677-84) The synthesis of phosphoramidite is as follows: synthesis 2-(2-((((6aR,8R,9R,9aR)-8-(6- Benzamide group -9H- Purine -9- base )-2,2,4,4- Tetraisopropyltetrahydrogen -6H- Furano [3,2-f][1,3,5,2,4] trioxadioxin (trioxadisilocin)-9- base ) Oxygen ) Methoxy ) Ethoxy ) Second -1- Ammonium formate (1-6) compound 1-1(A solution of 25.00 g, 67.38 mmol) in 20 mL of DMF was treated with pyridine (11 mL, 134.67 mmol) and tetraisopropyldisiloxane dichloride (22.63 mL, 70.75 mmol) at 10°C. handle. The resulting mixture was stirred at 25 °C for 3 h and quenched with 20% citric acid (50 mL). The aqueous layer was extracted with EtOAc (3X50 mL) and the combined organic layers were concentrated in vacuo. The crude residue was recrystallized from a mixture of MTBE and n-heptane (1:15, 320 mL) to obtain the compound as a white oily solid 1-2(37.20 g, 90%). compound 1-2(37.00 g, 60.33 mmol) in 20 mL of DMSO was added with AcOH (20 mL, 317.20 mmol) and Ac ₂O (15 mL, 156.68 mmol) treatment. The mixture was stirred at 25 °C for 15 h. The reaction was diluted with EtOAc (100 mL) and resolved with sat. K ₂CO ₃(50 mL) quenched. The aqueous layer was extracted with EtOAc (3X50 mL). The combined organic layers were concentrated and recrystallized from ACN (30 mL) to obtain the compound as a white solid. 1-3(15.65 g, 38.4%). compound 1-3(20.00 g, 29.72 mmol) in 120 mL of DCM was treated with Fmoc-amino-ethoxyethanol (11.67 g, 35.66 mmol) at 25°C. The mixture was stirred to obtain a clear solution and then filtered with 4Å molecular sieves (20.0 g), N- Treatment with iodosuccinimide (8.02 g, 35.66 mmol), and TfOH (5.25 mL, 59.44 mmol). The mixture was stirred at 30 °C until HPLC analysis indicated the compound 1-3until the consumption is >95%. The reaction was quenched with TEA (6 mL) and filtered. The filtrate was diluted with EtOAc and sat. NaHCO ₃(2X100 mL), sat. Na ₂SO ₃(2X100 mL), and water (2X100 mL) and concentrated in vacuo to give the crude compound as a yellow solid. 1-4(26.34 g, 93.9%), which was used directly in the next step without further purification. compound 1-4A solution of (26.34 g, 27.62 mmol) in a mixture of DCM/water (10:7, 170 mL) was treated with DBU (7.00 mL, 45.08 mmol) at 5 °C. The mixture was stirred at 5 to 25 °C for 1 h. The organic layer was then separated, washed with water (100 mL), and diluted with DCM (130 mL). The solution was divided into four parts and treated with butenedioic acid (7.05 g, 60.76 mmol) and 4Å molecular sieve (26.34g). The mixture was stirred for 1 h, concentrated, and recrystallized from a mixture of MTBE and DCM (5:1) to obtain the compound as a white solid. 1-6(14.74 g, 62.9%): ¹H NMR (400 MHz, d ₆ -DMSO)8.73(s, 1H), 8.58(s, 1H), 8.15-8.02(m, 2H), 7.65-7.60(m, 1H), 7.59-7.51(m, 2H), 6.52(s, 2H) , 6.15(s, 1H), 5.08-4.90(m, 3H), 4.83-4.78(m, 1H), 4.15-3.90(m, 3H), 3.79-3.65(m, 2H), 2.98-2.85(m, 6H), 1.20-0.95(m, 28H). synthesis (2R,3R,4R,5R)-5-(6- Benzamide group -9H- Purine -9- base )-2-(( pair (4- Methoxyphenyl )( phenyl ) Methoxy ) methyl )-4-((2-(2-[ Lipids ]- amide ethoxy ) Ethoxy ) Methoxy ) Tetrahydrofuran -3- base (2- Cyanoethyl ) diisopropylphosphoramidite (2-4a to 2-4e) compound 1-6(50.00 g, 59.01 mmol) in 150 mL of 2-methyltetrahydrofuran with ice-cold K ₂HPO ₄(6%, 100 mL) aqueous solution and brine (20%, 2X100 mL). The organic layer was separated and treated with caproic acid (10.33 mL, 82.61 mmol), HATU (33.66 g, 88.52 mmol), and DMAP (10.81 g, 147.52 mmol) at 0°C. The resulting mixture was warmed to 25 °C and stirred for 1 h. The solution was washed with water (2X100 mL), brine (100 mL), and concentrated in vacuo to give a crude residue. Silica gel flash chromatography (1:1 hexane/acetone) gave the compound as a white solid 2-1a(34.95 g, 71.5%). compound 2-1aA mixture of (34.95 g, 42.19 mmol) and TEA (9.28 mL, 126.58 mmol) in 80 mL of THF was treated dropwise with triethylamine trihydrofuride (20.61 mL, 126.58 mmol) at 10 °C. The mixture was warmed to 25 °C and stirred for 2 h. The reaction was concentrated, dissolved in DCM (100 mL) and washed with sat. NaHCO ₃(5X20 mL) and brine (50 mL). The organic layer was concentrated in vacuo to give crude compound 2-2a(24.72 g, 99%), which was used directly in the next step without further purification. compound 2-2a(24.72 g, 42.18 mmol) in 50 mL of DCM for N-Methylmarine (18.54 mL, 168.67 mmol) and DMTr-Cl (15.69 g, 46.38 mmol) treatment. The mixture was stirred at 25 °C for 2 h and washed with sat. NaHCO ₃(50 mL) quenched. The organic layer was separated, washed with water, and concentrated to obtain a crude syrup. Silica gel flash chromatography (1:1 hexane/acetone) gave the compound as a white solid 2-3a(30.05 g, 33.8 mmol, 79.9%). The compound was placed under a nitrogen atmosphere 2-3a(25.00 g, 28.17 mmol) in 50 mL of DCM for N-Methylmarin (3.10 mL, 28.17 mmol) and tetrazole (0.67 mL, 14.09 mmol) treatment. Bis(diisopropylamino)phosphine chloride (9.02 g, 33.80 mmol) was added dropwise to the solution and the resulting mixture was stirred at 25 °C for 4 h. The reaction was quenched with water (15 mL) and the aqueous layer was extracted with DCM (3X50 mL). The combined organic layers were treated with sat. NaHCO ₃(50 mL) and concentrated to obtain a crude solid, which was recrystallized from a mixture of DCM/MTBE/n-hexane (1:4:40) to obtain the compound as a white solid. 2-4a(25.52 g, 83.4%): ¹H NMR (400 MHz, d ₆ -DMSO)11.25(s, 1H), 8.65-8.60(m, 2 H), 8.09-8.02(m, 2H), 7.71(s, 1H), 7.67-7.60(m, 1H), 7.59-7.51(m , 2H), 7.38-7.34(m, 2H), 7.30-7.25(m, 7H), 6.85-6.79(m, 4H), 6.23-6.20(m, 1H), 5.23-5.14(m, 1H), 4.80 -4.69(m, 3H), 4.33-4.23(m, 2H), 3.90-3.78(m, 1H), 3.75(s, 6H), 3.74-3.52(m, 3H), 3.50-3.20(m, 6H) , 3.14-3.09(m, 2H), 3.09(s, 1H), 2.82-2.80(m, 1H), 2.65-2.60(m, 1H), 2.05-1.96(m, 2H), 1.50-1.39(m, 2H), 1.31-1.10(m, 14H), 1.08-1.05(m, 2H), 0.85-0.79(m, 3H); ³¹P NMR (162 MHz, d ₆ -DMSO)149.43, 149.18. compound 2-4b , 2-4c , 2-4d ,and 2-4eThe above compounds are used 2-4aPrepared by similar procedures. Obtain the compound as a white solid 2-4b(25.50 g, 85.4%): ¹H NMR (400 MHz, d ₆ -DMSO)11.23(s, 1H), 8.65-8.60(m, 2 H), 8.05-8.02(m, 2H), 7.73-7.70(m, 1H), 7.67-7.60(m, 1H), 7.59-7.51 (m, 2H), 7.38-7.34(m, 2H), 7.30-7.25(m, 7H), 6.89-6.80(m, 4H), 6.21-6.15(m, 1H), 5.23-5.17(m, 1H) , 4.80-4.69(m, 3H), 4.40-4.21(m, 2H), 3.91-3.80(m, 1H), 3.74(s, 6H), 3.74-3.52(m, 3H), 3.50-3.20(m, 6H), 3.14-3.09(m, 2H), 3.09(s, 1H), 2.83-2.79(m, 1H), 2.68-2.62(m, 1H), 2.05-1.97(m, 2H), 1.50-1.38( m, 2H), 1.31-1.10(m, 18H), 1.08-1.05(m, 2H), 0.85-0.78(m, 3H); ³¹P NMR (162 MHz, d ₆ -DMSO)149.43, 149.19. Obtain compound as off-white solid 2-4c(36.60 g, 66.3%): ¹H NMR (400 MHz, d ₆ -DMSO)11.22(s, 1H), 8.64-8.59(m, 2H), 8.05-8.00(m, 2H), 7.73-7.70(m, 1H), 7.67-7.60(m, 1H), 7.59-7.51( m, 2H), 7.38-7.34(m, 2H), 7.30-7.25(m, 7H), 6.89-6.80(m, 4H), 6.21-6.15(m, 1H), 5.25-5.17(m, 1H), 4.80-4.69(m, 3H), 4.40-4.21(m, 2H), 3.91-3.80(m, 1H), 3.74(s, 6H), 3.74-3.50(m, 3H), 3.50-3.20(m, 6H ), 3.14-3.09(m, 2H), 3.09(s, 1H), 2.83-2.79(m, 1H), 2.68-2.62(m, 1H), 2.05-1.99(m, 2H), 1.50-1.38(m , 2H), 1.33-1.12(m, 38H), 1.08-1.05(m, 2H), 0.86-0.80(m, 3H); ³¹P NMR (162 MHz, d ₆ -DMSO)149.42, 149.17. Obtain compound as off-white solid 2-4d(26.60 g, 72.9%): ¹H NMR (400 MHz, d ₆ -DMSO)11.22(s, 1H), 8.64-8.59(m, 2H), 8.05-8.00(m, 2H), 7.73-7.70(m, 1H), 7.67-7.60(m, 1H), 7.59-7.51( m, 2H), 7.38-7.33(m, 2H), 7.30-7.25(m, 7H), 6.89-6.80(m, 4H), 6.21-6.15(m, 1H), 5.22-5.17(m, 1H), 4.80-4.69(m, 3H), 4.40-4.21(m, 2H), 3.91-3.80(m, 1H), 3.74(s, 6H), 3.74-3.52(m, 3H), 3.50-3.20(m, 6H ), 3.14-3.09(m, 2H), 3.09(s, 1H), 2.83-2.79(m, 1H), 2.68-2.62(m, 1H), 2.05-1.99(m, 2H), 1.50-1.38(m , 2H), 1.35-1.08(m, 38H), 1.08-1.05(m, 2H), 0.85-0.79(m, 3H); ³¹P NMR (162 MHz, d ₆ -DMSO)149.47, 149.22. Obtain the compound as a white solid 2-4e(38.10 g, 54.0%): ¹H NMR (400 MHz, d ₆ -DMSO)11.21(s, 1H), 8.64-8.59(m, 2H), 8.05-8.00(m, 2H), 7.73-7.70(m, 1H), 7.67-7.60(m, 1H), 7.59-7.51( m, 2H), 7.38-7.34(m, 2H), 7.30-7.25(m, 7H), 6.89-6.80(m, 4H), 6.21-6.15(m, 1H), 5.23-5.17(m, 1H), 4.80-4.69(m, 3H), 4.40-4.21(m, 2H), 3.91-3.80(m, 1H), 3.73(s, 6H), 3.74-3.52(m, 3H), 3.47-3.22(m, 6H ), 3.14-3.09(m, 2H), 3.09(s, 1H), 2.83-2.79(m, 1H), 2.68-2.62(m, 1H), 2.05-1.99(m, 2H), 1.50-1.38(m , 2H), 1.35-1.06(m, 46H), 1.08-1.06(m, 2H), 0.85-0.77(m, 3H); ³¹P NMR (162 MHz, d ₆ -DMSO)149.41, 149.15. Oligomers were analyzed using ion-exchange high performance liquid chromatography (IE-HPLC) on an Amersham Source 15Q column (1.0 cm × 25 cm; Amersham Pharmacia Biotech) using a 15 min step Purify using step-linear gradient. The gradient varied from 90:10 buffer A:B to 52:48 buffer A:B, where buffer A was 100 mM Tris pH 8.5 and buffer B was 100 mM Tris pH 8.5, 1 M NaCl. Samples were monitored at 260 nm and peaks corresponding to full-length oligonucleotide species were collected, pooled, desalted on a NAP-5 column, and freeze-dried. The purity of each oligomer was determined by capillary electrophoresis (CE) on a Beckman PACE 5000 (Beckman Coulter, Inc.; Fullerton, CA). CE capillaries have an inner diameter of 100 μm and contain ssDNA 100R gel (Beckman-Coulter). Typically, approximately 0.6 nmole of oligonucleotide is injected into a capillary, run in an electric field of 444 V/cm, and detected by UV absorbance at 260 nm. Denatured Tris-Borate-7 M-urea electrophoresis buffer was purchased from Beckman-Coulter. The oligonucleotides obtained were at least 90% pure as assessed by CE and were used in the following examples. Compound identification was performed using a matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometer in the Voyager DE™ Biospectometry Work Station (Applied Biosystems; Foster City, CA). Verify with the manufacturer's recommended procedures. The relative molecular weights of all oligomers were obtained and were usually within 0.2% of the expected molecular weight. Preparation of doubletsResuspend the single-stranded RNA oligos (e.g., at a concentration of 100 μM) in duplex buffer consisting of 100 mM potassium acetate, 30 mM HEPES (pH 7.5). Complementary sense and antisense strands are mixed in equimolar amounts to yield, for example, a 50 μM final solution of the duplex. Samples were heated to 100°C in RNA buffer (IDT) for 5' and allowed to cool to room temperature before use. Store RNAi oligonucleotides at -20°C. Single-stranded RNA oligos are stored freeze-dried or stored in nuclease-free water at -80°C. The synthetic methods described in this article were used to generate Example 4 , 5 , 7 ,and 8Lipid-conjugated oligonucleotides as described in . Example 2 : GalNAc- Combined GFAP RNAi Intrathecal and intracerebroventricular administration of oligonucleotides inhibits mice in a concentration-dependent manner in vivo Gfap Glial fibrillary acidic protein (GFAP) encodes an intermediate filament protein found primarily in stellate cells of the CNS. GFAPSufficient attenuation demonstrates the ability of the oligonucleotides described herein to reduce the expression of the target gene expressed in stellate cells. To assess the in vivo reduction of RNAi oligonucleotides GfapThe ability to perform, using e.g. Example 1Oligonucleotides were synthesized as described in to generate double-stranded RNAi oligonucleotides containing a gapped four-ring GalNAc-bound structure (referred to herein as "GalNAc-bound GFAPOligonucleotide (GalNAc-conjugated GFAPoligonucleotide)" or "GalNAc- GFAPOligonucleotide"), which has a 36-mer follower strand and a 22-mer leader strand. Further, the nucleotide sequences comprising the follower and leader strands have different patterns of modified nucleotides and phosphorothioate linkages. Each of the three nucleotides that comprise the four loops binds to the GalNAc moiety (CAS#14131-60-3). The modification mode of each stock is explained as follows: Cross to: Modifier keys: surface 1 symbol Modification / keying [MePhosphonate-4O-mUs] Nucleotide modified with 4'-O-monomethylphosphonate-2'-O-methyl [ademX-GalNAc] GalNAc attaches to nucleotides [mXs] 2'- O -methyl modified nucleotides with phosphorothioate linkages to adjacent nucleotides [fXs] 2'-fluoro modified nucleotides with phosphorothioate linkages to adjacent nucleotides [mX] 2'- O -methyl modified nucleotides with phosphodiester linkages to adjacent nucleotides [fX] 2'-fluoro modified nucleotides with phosphodiester linkages to adjacent nucleotides [AdemX-C16] C16 lipid attached to nucleotides [AdemX-C16s] C16 lipids attached to nucleotides with phosphorothioate linkages to adjacent nucleotides i. Intrathecal administrationEvaluate GFAP-1477 (e.g. surface 2(shown in ) decreases in the central nervous system (CNS) of mice via intrathecal (i.t.) administration Gfapability. Specifically, mice were administered 10, 32, 100, 320, or 1000 µg of GFAP-1477 formulated in artificial cerebrospinal fluid (aCSF) via i.t. lumbar injection. Animals were sacrificed on day 7 after intrathecal injection. RNA was extracted from CNS tissue and assayed by qPCR in mice GfapmRNA levels (for endogenous housekeeping genes Rpl23standardization). Murine using the PrimeTime™ qPCR Probe Assay (IDT) GfapThe level of mRNA, which consists of a pair of primers and a fluorescently labeled 5' nuclease probe, which is specific for mice GfapmRNA. Rats in treated mouse samples GfapThe percentage of mRNA residue was calculated using 2 ^-ΔΔCt("δ-δ Ct") method (Livak and Schmittgen (2001) METHODS 25:402-408). Observed in CNS tissues GfapPerformance decreased in a dose-dependent manner. Specifically, such as Figure 1A , Figure 1B , Figure 1C , and pictures 1FAs shown in respectively, in samples from the cervical spinal cord, thoracic spinal cord, lumbar spinal cord, and cerebellum GfapPerformance is reduced by approximately 50% or more. like Figure 1D and pictures 1Eshown respectively, observed in the frontal cortex and hippocampus GfapPerformance degradation is less. Figure 2Shown in ₅₀value. Taken together, this data shows that a GalNAc-bound tetracyclic GFAPTargeting oligonucleotide inhibits in several tissues of the CNS in a dose-dependent manner following i.t. administration GFAPPerformance. ii. intracerebroventricular administrationGFAP-1477 was also assessed to reduce the effects of GFAP-1477 in the central nervous system (CNS) in mice via intracerebroventricular (i.c.v) administration. Gfapability. Specifically, mice were administered 10, 32, 100, or 300 µg of GFAP-1477 formulated in aCSF via i.c.v. Animals were sacrificed on day 7 after i.c.v. injection. RNA was extracted from CNS tissue as described above. Observed in CNS tissue GfapPerformance is reduced in a dose-dependent manner. Specifically, such as Figure 3A To the picture 3DAs shown in respectively, in the samples of frontal cortex, brainstem, hippocampus, and lumbar spinal cord, GfapPerformance is reduced by approximately 50% or more. Figure 4Shown in ₅₀value. Taken together, this data shows that a GalNAc-bound tetracyclic GFAPTargeted oligonucleotides inhibited in several tissues of the CNS in a dose-dependent manner when administered via i.c.v. GFAPPerformance. Example 3 : Synthesis of tetracyclic rings RNAi Oligonucleotides - Lipid conjugatesThe lipid-conjugated tetracyclic oligonucleotides described herein can be synthesized using synthetic methods later detailed in PCT Application No. PCT/US2021/42469. Specifically, oligonucleotides can be synthesized using a post-synthetic conjugation approach such as that shown below. plan 1Synthesis of RNAi oligonucleotide-lipid conjugates with single lipids (linear and branched) bound to tetracyclic rings. Post-synthetic conjugation can be achieved by amide coupling reactions. R ₁The COOH group represents fatty acids C8:0, C10:0, C11:0, C12:0, C14:0, C16:0, C17:0, C18:0, C18:1, C18:2, C22:5, C22 :0, C24:0, C26:0, C22:6, C24:1, diyl C16:0, or diyl C18:1. An exemplary R1 structure is provided below: synthesis justice unit 1and Antisense stocks 1It is prepared by solid phase synthesis. synthesis Combined Justice Shares 1ato 1i. Combined Justice Shares 1aSynthesized by post-synthetic combining methods .A solution of caprylic acid (0.58 mg, 4 umol) in DMA (0.75 mL) was treated with HATU (1.52 mg, 4 umol) in Eppendorf tube 1 at rt. In Ebende tube 2, place the oligonucleotide justice unit 1(10.00 mg, 0.8 umol) in H ₂A solution in O (0.25 mL) was treated with DIPEA (1.39 uL, 8 umol). Add the solution in Ebende Tube 1 to Ebende Tube 2 and mix using a ThermoMixer at rt. After indicating completion of the reaction by LC-MS analysis, the reaction mixture was diluted with 5 mL of water and passed through a reverse-phase XBridge C18 column using 100 mM TEAA in ACN and H ₂Purify using a gradient of 5 to 95% in O. The product fractions were concentrated under reduced pressure using Genevac. The combined residual solvents were dialyzed against water (1X), brine (1X), and water (3X) using an Amicon® Ultra-15 Centrifugal (3K). The Amicon membrane was washed with water (3 × 2 mL), and then the combined solvents were freeze-dried to obtain Combined Justice Shares 1aamorphous white solid (6.43 mg, 64% yield). Combined Justice Shares 1b to 1iSystem and synthesis Combined Justice Shares 1aA similar procedure was described and obtained in 42% to 69% yield. bond double body 1a to 1jwill Combined Justice Shares 1a(10 mg by weight) was dissolved in 0.5 mL of deionized water to prepare a 20 mg/mL solution. will Antisense stocks 1(10 mg, measured by OD) was dissolved in 0.5 mL of deionized water to prepare a 20 mg/mL solution, which was used for titration of bound sense strands and quantification of duplex volume. Based on the calculation of the molar amount of the combined sense and antisense shares, the required Antisense stocks 1Add section to Combined Justice Shares 1ain solution. The resulting mixture was stirred at 95 °C for 5 min and allowed to cool to rt. The adhesion progress was monitored by ion exchange HPLC. Based on the gluing process, several further Antisense stocks 1Partially bonded, purity >95%. The solution was freeze-dried to obtain double body 1a(C8)And the amount is calculated based on the molar amount of antisense strand consumed in adhesion. double body 1b to 1isystem use and bonding double body 1a(C8)Prepared by the same procedure as described. Scheme 1-2 below illustrates the synthesis of a gapped tetracyclic GalXC conjugate with a single lipid on the loop. Post-synthetic conjugation is achieved through Cu-catalyzed alkyne-azide cycloaddition reaction. Justice Unit 1Band Antisense stocks 1BIt is prepared by solid phase synthesis. synthesis Combined Justice Shares 1j. In Ebende tube 1, dissolve oligonucleotide (10.00 mg, 0.8 umol) in DMA/H ₂A solution in a 3:1 mixture of O (0.5 mL) was treated with lipid linker azide (11.26 mg, 4 umol). In Ebende tube 2, dissolve CuBr dimethyl sulfide (1.64 mg, 8 umol) in ACN (0.5 mL). Combine the two solutions by adding N ₂Degas by bubbling through for 10 min. Then CuBrSMe ₂The ACN solution was added to tube 1 and the resulting mixture was stirred at 40 °C. After indicating completion of the reaction by LC-MS analysis, the reaction mixture was diluted with 0.5 M EDTA (2 mL) and dialyzed against water (2X) using Amicon® Ultra-15 Centrifugal (3K). The reaction crude product was passed through a reverse phase XBridge C18 column using 100 mM TEAA in ACN (added 30% IPA) and H ₂Purify using a gradient of 5 to 95% in O. The product fractions were concentrated under reduced pressure using Genevac. The combined residual solvents were dialyzed against water (1X), brine (1X), and water (3X) using an Amicon® Ultra-15 Centrifugal (3K). The Amicon membrane was washed with water (3 × 2 mL) and the combined solvents were freeze-dried to obtain Combined Justice Shares 1jamorphous white solid (6.90 mg, 57% yield). double body 1j( PEG2K- Dihydroxyl C18)system use and bonding double body 1a(C8)Prepared by the same procedure as described. Schemes 1-3 below illustrate the synthesis of gapped tetracyclic GalXC conjugates with double lipids on the loop using post-synthetic conjugation methods. justice unit 2and Antisense stocks 2It is prepared by solid phase synthesis. Combined Justice Shares 2aand 2bSystem used with Combined Justice Shares 1aSimilar procedure, but prepared with 10 eq of lipids, 10 eq of HATU, and 20 eq of DIPEA. double body 2a(2XC11)and 2b(2XC22)system use and bonding double body 1a(C8)Prepared by the same procedure as described. Schemes 1-4 below illustrate the synthesis of fully phosphorothioated backbone-loop GalXC conjugated to a single lipid using a post-synthetic conjugation approach. justice unit 3and Antisense stocks 3It is prepared by solid phase synthesis. Combined Justice Shares 3asystem use and synthesis Combined Justice Shares 1aA similar procedure was prepared and obtained in 65% yield. double body 3a(PS-C22)system use and bonding double body 1a(C8)Prepared by the same procedure as described. Schemes 1-5 below illustrate the use of post-synthetic conjugation methods to synthesize short sense strands of GalXC conjugated to a single lipid. justice unit 4and Antisense stocks 4It is prepared by solid phase synthesis. Combined Justice Shares 4asystem use and synthesis Combined Justice Shares 1aA similar procedure was prepared and obtained in 74% yield. double body 4a(SS-C22)system use and bonding double body 1a(C8)Prepared by the same procedure as described. Schemes 1-6 below illustrate examples of solid-phase synthesis of gapped tetracyclic GalXC conjugates with lipid(s) on the loop. Synthetic combination of justice shares 6 . Combined Justice Shares 6It was prepared by solid-phase synthesis using a commercially available oligonucleotide synthesizer. The oligonucleotide uses a 2'-modified nucleoside phosphoramidite (such as 2'-F, or 2'-OMe) and a 2'-diethoxymethanol-linked fatty acid amide nucleoside phosphite Amide synthesis. Oligonucleotide synthesis was performed in the 3' to 5' direction on a solid support using standard oligonucleotide synthesis protocols. 5-Ethylthio-1H-tetrazole (ETT) is used as the activator of the coupling reaction. Iodine solution is used in the phosphite attempts oxidation reaction. 3-(Dimethylaminomethylene)amino-3H-1,2,4-dithiazole-3-thione (DDTT) is used to form phosphorothioate linkages. The synthesized oligonucleotides were treated with concentrated aqueous ammonium for 10 h. Ammonia was removed from the suspension and solid support residue was removed by filtration. The crude oligonucleotides were treated with TEAA, analyzed, and purified by strong anion exchange high performance liquid chromatography (SAX-HPLC). Fractions were combined and dialyzed against water (3X), saline (1X), and water (3X) using an Amicon® Ultra-15 Centrifugal (3K). The remaining solvent is then freeze-dried to obtain the desired Combined Justice Shares 6. double body 6system use and bonding double body 1a(C8)Prepared by the same procedure as described. the following plan 1-7Shown is the synthesis of GalXC conjugated to a single lipid at the 5' end using a post-synthetic conjugation approach. Synthetic Combination of Justice Shares 10a Combined Justice Shares 10asystem use and synthesis Combined Justice Shares 5Obtained by the same method or substantially similar method. double body 10aSynthetic Examples double body 10asystem use and synthesis double body 5Obtained by the same method or substantially similar method. Example 4 : GFAP- Targeted oligonucleotides show long duration of action via intrathecal and intracisternal administration (Long-term Duration of Action)In this example, the duration of action was determined using oligonucleotides with different targeting ligands. Specifically, it will be as follows: Example 2GalNAc-binding as described in GfapOligonucleotides, and Example 1C16-conjugated generated by the method described in GfapOligonucleotides (the sequences of which are provided in surface 3Medium) administered to rats for long-term studies. The C16 lipid moiety is bound to position 28 of the sense strand. Four-loop RNAi oligonucleotide modification pattern Cross to: Modifier keys: surface 1 i. Intrathecal administrationThe above GalNAc-conjugated and lipid-conjugated GfapOligonucleotides were administered to rats via intrathecal (i.t.) injection for assessment GfapA long-lasting period of reduced performance. Specifically, Sprague-Dawley rats (250 g) were administered a single 1000 µg dose of 1000 µg in aCSF via i.t. injection. surface 3Among the oligonucleotides. Target weakening was assessed at weeks 8, 12, and 23 after i.t. injection. RNA was extracted from tissue samples of the prefrontal cortex, sensory cortex, striatum, hippocampus, periaqueductal gray, cerebellum, brainstem, and spinal cord (SC) segments 1 to 8 (SC1 to SC8). , determined by qPCR in rats GfapmRNA levels, such as Example 2described in. Rats GfapThe levels of mRNA were measured using the PrimeTime™ qPCR Probe Assay (IDT). Over the course of the study (i.e., at weeks 8, 12, and 23), C16-binding oligonucleotides were shown to be present in the brainstem and SC1 to SC8. GfapPerformance is reduced by approximately 50% or more ( Figure 5A). In contrast, in all brain tissues sampled, using GalNAc-conjugated oligonucleotides GfapLess performance degradation ( Figure 5B). This data shows that stellate cell target mRNA ( For example, GFAP)of long-term continued suppression. ii. intracisternal administrationwill surface 3C16-binding as described in GFAPOligonucleotides were administered intracisternally (i.c.m.) to rats for assessment GfapA long-lasting period of reduced performance. Specifically, Stewart rats were administered a single 1000 µg dose of oligonucleotide formulated in aCSF via i.c.m injection. Assessments at 4 and 12 weeks after injection Gfapits performance. RNA was extracted from tissue samples of prefrontal cortex, sensory cortex, hippocampus, hypothalamus, cerebellum, brainstem, cervical spinal cord, and lumbar spinal cord, and assayed by qPCR in rats. GfapmRNA levels, such as Example 2described in. Observed in all brain regions at 4 weeks after i.c.m. injection GfapPerformance is reduced by about 50% or more, and in the cerebellum, brainstem, cervical spinal cord, and lumbar spinal cord, about 50% or more GfapReduced performance was maintained until week 12 after i.c.m. injection ( Figure 6). Investing in the second animal group surface 3The same C16 lipid in - combined with GfapOligonucleotides are assessed through weeks 26 and 39 GfapDecreased expression of mRNA. Rats were administered a single dose of oligonucleotide as described above. RNA was extracted from tissue samples of frontal cortex, striatum, sensory cortex, hippocampus, hypothalamus, circumaqueductal gray matter, cerebellum, brainstem, and spinal cord (SC) segments 1 to 8 (SC1 to SC8). , determined by qPCR in rats GfapmRNA levels, such as Example 2described in. At 26 weeks after dosing, it was observed in the cerebellum, brainstem, and SC1 to SC5 GfapPerformance is reduced by approximately 50% or more ( Figure 7A). By week 39, observed in the cerebellum, brainstem, and SC1 to SC7 GfapPerformance is reduced by approximately 25 to 50% ( Figure 7B). This data shows that stellate cell target mRNA ( For example, GFAP)of long-term continued suppression. Example 5 : In the Fourth Ring Road RNAi oligonucleotide lipids - Positional effects of lipid binding on in vivo activity of conjugates in the central nervous systemThe ability of RNAi oligonucleotide-lipid conjugates containing tetraloops to reduce mRNA expression in CNS stellate cells was evaluated in vivo. C16-Combined GfapOligonucleotide systems such as Example 1generated as described in . Specifically, the C16 lipid binds to the sense strand at one of positions (P) 1, 4, 8, 12, 13, 18, 20, 23, 28, 29, and 30, as shown in the modification pattern below. Show. The unmodified sense and antisense strands are provided in SEQ ID NO: 3 and 4 respectively, while the modified strands are shown in surface 4middle. Four-loop RNAi oligonucleotide modification pattern: P1 Justice shares: P4 Justice shares: P8 Justice shares: P12 Justice shares: P13 Justice shares: P18 Justice shares: P20 Justice shares: P23 Justice shares: P28 Justice shares: P29 Justice shares: P30 Justice shares: Each of P1, P4, P8, P12, P13, P18, P20, P23, P28, P29, and P30 was hybridized to an antisense strand with the following modification pattern: Modifier keys: surface 1 in order to evaluate surface 4For the oligonucleotides in, 6- to 8-week-old C57BL/6 female mice were treated with 300 µg of RNAi oligonucleotides in artificial cerebrospinal fluid via intrathecal (i.t.) lumbar injection. Target performance was assessed on day 7 after injection. RNA was extracted from tissue samples of the lumbar spinal cord, medulla oblongata, cerebellum, hypothalamus, hippocampus, and frontal cortex, and assayed by qPCR in mice. GfapmRNA levels, such as Example 2described in. In the lumbar spinal cord, all oligonucleotides were reduced GfapmRNA expression( Figure 8A). Oligonucleotides binding to C16 lipids at P1, P4, P13, P18, P20, P23, P29, or P30 reduce mRNA expression in the medulla oblongata ( Figure 8B). In the cerebellum, expression was reduced by oligonucleotides binding to C16 lipids at P4, P23, or P29 ( Figure 8C). Oligonucleotides that bind to C16 lipids at P1, P4, P12, P13, P18, P20, P23, P28, P29, or P30 reduce the GfapmRNA expression( Figure 8D). Slight weakening was observed in the hippocampus and frontal cortex (respectively Figure 8E and pictures 8F). In summary, C16-conjugated tetracyclic oligonucleotides reduce inflammatory response in several CNS tissues after administration to the CNS via i.t. waist injection. GfapPerformance. Example 6 :Synthetic lipid - Blunt-ended oligonucleotideThe following schematic illustrates the synthesis of a blunt-ended oligonucleotide with a C16-lipid at the 5'-end. Lipid-conjugated blunt-ended oligonucleotides described herein may be synthesized using synthetic methods later detailed in PCT Application No. PCT/US2021/42469. Specifically, oligonucleotides can be synthesized using post-synthesis conjugation methods such as those illustrated below. In Ebende tube 1, a solution of palmitic acid in DMA was treated with HATU at rt. In Ebende tube 2, place the oligonucleotide sense strand in H ₂The solution in O was treated with DIPEA. Add the solution in Ebende Tube 1 to Ebende Tube 2 and mix using a ThermoMixer at rt. After indicating completion of the reaction by LC-MS analysis, the reaction mixture was diluted with 5 mL of water and passed through a reverse-phase XBridge C18 column using 100 mM TEAA in ACN and H ₂Purify using a gradient of 5 to 95% in O. The product fractions were concentrated under reduced pressure using Genevac. The combined residual solvents were dialyzed against water (1X), brine (1X), and water (3X) using an Amicon® Ultra-15 Centrifugal (3K). The Amicon membrane was washed with water (3 × 2 mL) and the combined solvents were freeze-dried to give an amorphous white solid. Example 7 : at the blunt end RNAi Positional effects of lipid binding of oligonucleotides on in vivo activity in the central nervous systemRNAi oligonucleotide conjugates that bind to lipids at different locations on the sense strand were evaluated for their ability to reduce stellate cell target expression in the CNS. RNAi oligonucleotides binding to C16 lipids were generated as described above. Specifically, a protein with a blunt end at the 3' end and a 2 nucleotide overhang at the 5' end was generated, with the C16 lipid bound to the sense strand at positions (P) 1, 4, 8, 12, 13 , 18 and 20 oligonucleotides, as shown in the modification pattern below. Comparison includes from Example 5Selected lipid-bound tetracyclic oligonucleotides (P1, P4, P23, and P28). The unmodified sense and antisense strands are provided in SEQ ID NO: 19 and 4 respectively, while the modified strands are shown in surface 5middle. Blunt-ended RNAi oligonucleotide modification patterns: P1 Justice shares: P4 Justice shares: P8 Justice shares: P12 Justice shares: P13 Justice shares: P18 Justice shares: P20 Justice shares: Each of P1, P4, P8, P12, P13, P18, and P20 was hybridized to an antisense strand with the following modification pattern: Modifier keys: surface 1 in order to evaluate surface 5The blunt-end oligonucleotide-lipid conjugate in 6- to 8-week-old C57BL/6 female mice was administered via intrathecal (i.t.) waist injection with 500 µg of lipid in artificial cerebrospinal fluid (aCSF). -Conjugated blunt-ended RNAi oligonucleotide treatment. Control animals were injected with aCSF only. Target performance was assessed on day 7 after injection. RNA was extracted from tissue samples of the lumbar spinal cord, medulla oblongata, cerebellum, hypothalamus, hippocampus, and frontal cortex, and assayed by qPCR in mice. GfapmRNA levels (for endogenous housekeeping genes Rpl23Standardized, as indicated). use Example 2Rats were determined as described in Gfaplevels of mRNA. In a sample of several tissues from the CNS, GfapmRNA expression is reduced by approximately 50% or more ( Figure 9A To the picture 9F). Specifically, in the lumbar spinal cord and medulla oblongata, all lipid binding sites were reduced GfapmRNA( Figure 9A and pictures 9B). In the cerebellum, oligonucleotides that bind to C16 lipids at P4, P12, P13, P18, or P20 reduce mRNA expression ( Figure 9C). In the hypothalamus, performance was reduced by oligonucleotides binding to C16 lipids at P1, P4, P12, P13, P18, or P20 ( Figure 9D). Observed in the hippocampus and frontal cortex GfapSlight inhibition of mRNA (respectively Figure 9E and pictures 9F). For each brain region, the lipid-conjugated tetracyclic oligonucleotides tested all showed consistent Example 5Similar performance degradation is shown in . Overall, several lipid-binding sites in blunt-ended oligonucleotides are successful inhibitors of target mRNAs in the CNS. Compare from Example 5and the remaining percentage of mRNA for the experiments described in this example. Specifically, the data are summarized in Figure 10A To the picture 10F, which shows the potency of tetracyclic and blunt-end oligonucleotides across brain regions and lipid locations. Example 8 : C16- Combined GFAP Blunt-ended oligonucleotides inhibit mice in a concentration-dependent manner in vivo Gfap Assessing the effect of blunt-ended oligonucleotides that bind to C16 lipids at position 1 when administered i.t. in a concentration-dependent manner on reducing the central nervous system (CNS) in mice Gfapability. Specifically, mice were administered 3, 10, 30, 100, or 300 µg of GFAP-1477 (SEQ ID NO: 20 (sense) and SEQ ID NO: 18 (anti) formulated in aCSF via i.t. injection. Loyal shares)). Animals were sacrificed on day 7 or 28 after i.t. injection. like Example 2RNA was extracted from liver tissue as described. Observed in several CNS tissues GfapPerformance decreases in a concentration-dependent manner. Specifically, on day 7 after injection, expression decreased in the hypothalamus, cerebellum, brainstem, and lumbar spinal cord ( Figure 11A). In the same tissue, this decrease was maintained until day 28, indicating long-term suppression after a single administration of blunt-ended oligonucleotide ( Figure 11B). Taken together, this data demonstrates the long-term efficacy of lipid-conjugated blunt-ended oligonucleotides in inhibiting stellate cell target mRNA.

[ Figure 1A ] to [ Figure 1F ] provide graphs illustrating the concentration response of GalNAc-binding oligonucleotides targeting the stellate cell-specific gene GFAP . The percentage (%) of murine Gfap mRNA remaining in the following regions of mice after treatment with GalNAc-conjugated Gfap oligonucleotides ( Table 2 ) was determined: cervical spinal cord ( Fig. 1A ), thoracic spinal cord ( Fig. 1B ) , lumbar spinal cord ( Figure 1C ), frontal cortex ( Figure 1D ), hippocampus ( Figure 1E ), and cerebellum ( Figure 1F ). Mice were treated with 10, 32, 100, 320, or 1000 µg of GalNAc-conjugated Gfap oligonucleotide in artificial cerebrospinal fluid (aCSF) via intrathecal injection into the lumbar spine. Seven (7) days after intrathecal injection, Gfap mRNA levels were normalized to ribosomal protein L23 (RPL23) mRNA and overall performance determined between tissue types relative to control mice treated with aCSF. [ Figure 2 ] Provides a graph depicting the average percentage (%) of murine Gfap mRNA remaining in different tissues of the central nervous system (CNS) and calculated for each CNS tissue based on the results in Figures 1A to 1F EC ₅₀ (ED ₅₀ ). [ Figure 3A ] to [ Figure 3D ] provide graphs illustrating the concentration response of GalNAc-bound Gfap oligonucleotides. The percentage (%) of murine Gfap mRNA remaining in the following regions of mice after treatment with GalNAc-conjugated Gfap oligonucleotides ( Table 2 ) was determined: frontal cortex ( Fig. 3A ), brain stem ( Fig. 3B) ), hippocampus ( Figure 3C ), and lumbar spinal cord ( Figure 3D ). Mice were treated via intracerebroventricular (icv) injection with 10, 32, 100, or 300 µg of oligonucleotides in artificial cerebrospinal fluid (aCSF). Seven (7) days after icv injection, Gfap mRNA levels were normalized to ribosomal protein L23 (RPL23) mRNA and overall performance determined between tissue types relative to control mice treated with aCSF. [ Figure 4 ] A graph is provided illustrating the average percentage (%) of remaining murine Gfap mRNA in different tissues of the CNS based on the results in Figures 3A to 3D and the EC ₅₀ (ED _{50 )} calculated for each CNS tissue. ). [ Figure 5A ] to [ Figure 5B ] provide diagrams illustrating the use of C16 lipid-conjugated Gfap tetraloop oligonucleotide ( Figure 5A ) or the use of GalNAc-conjugated "GalXC" Gfap tetraloop oligonucleotide. Percentage (%) of rat Gfap mRNA re-exploited in central nervous system tissue of rats after acid treatment ( Fig. 5B ). Rats were treated with 1000 µg of the Gfap oligonucleotides indicated in Table 3 formulated in artificial cerebrospinal fluid (aCSF) via intrathecal injection into the lumbar spine. Gfap mRNA levels were normalized to peptidylprolyl Isomerase B (Ppib) mRNA at 8, 12, or 23 weeks post-dose and determined between tissue types relative to those treated with aCSF Overall performance of treated control rats. [ Figure 6 ] A graph is provided showing the percentage (%) of rat Gfap mRNA recovered in rat central nervous system tissue after treatment with C16 lipid-conjugated Gfap tetracyclic oligonucleotide. Rats were treated via intracisternal injection with 1000 µg of the Gfap lipid-conjugated tetracyclic oligonucleotides indicated in Table 3 formulated in artificial cerebrospinal fluid (aCSF). Levels of Gfap mRNA were normalized to peptidyl prolinyl isomerase B (Ppib) mRNA at 4 or 12 weeks post-dose and determined between tissue types relative to control rats treated with aCSF. Overall performance. [ Figures 7A - 7B ] Provided are graphs illustrating the percentage (%) of rat Gfap mRNA recovered in rat central nervous system tissue following treatment with C16 lipid-conjugated Gfap tetracyclic oligonucleotide. Rats were treated via intracisternal injection with 1000 µg of the Gfap lipid-conjugated tetracyclic oligonucleotides indicated in Table 3 formulated in artificial cerebrospinal fluid (aCSF). Levels of Gfap mRNA were normalized to peptidylprolinyl isomerase B (Ppib) mRNA at either week 26 ( Fig. 7A ) or week 39 ( Fig. 7B ) after administration and determined between tissue types. Overall performance relative to control rats treated with aCSF. [ Figure 8A ] to [ Figure 8F ] provide graphs illustrating the remaining in the following regions of mice after treatment with lipid Gfap tetracyclic oligonucleotides with binding at the nucleotide positions indicated on the x-axis. Percentage (%) of mouse Gfap mRNA: lumbar spinal cord ( Figure 8A ), medulla oblongata ( Figure 8B ), cerebellum ( Figure 8C ), hypothalamus (Figure 8D ), hippocampus ( Figure 8E), and frontal cortex ( Figure 8E ) 8F ). Mice were treated via intrathecal (it) injection with 300 µg of the Gfap lipid-conjugated tetracyclic oligonucleotides indicated in Table 4 formulated in artificial cerebrospinal fluid (aCSF). On day 7 post-dose, Gfap mRNA levels were normalized to ribosomal protein L23 (RPL23) mRNA and overall performance determined between tissue types relative to control mice treated with aCSF. [ Figure 9A ] to [ Figure 9F ] provide graphs illustrating the following regions in mice after treatment with lipid Gfap blunt-ended or tetracyclic oligonucleotides with binding at the nucleotide positions indicated on the x-axis Percentage (%) of mouse Gfap mRNA remaining in: lumbar spinal cord ( Figure 9A ), medulla oblongata ( Figure 9B ), cerebellum ( Figure 9C ), hypothalamus ( Figure 9D ), hippocampus ( Figure 9E ), and frontal lobe cortex ( Fig. 9F ). Mice were treated with 300 µg of the C16 lipid-conjugated Gfap oligonucleotides indicated in Table 5 formulated in artificial cerebrospinal fluid (aCSF) via intrathecal injection into the lumbar spine. Seven (7) days after intrathecal injection, Gfap mRNA levels were normalized to ribosomal protein L23 (RPL23) mRNA and overall performance determined between tissue types relative to control mice treated with aCSF. [ FIG. 10A ] to [ FIG. 10F ] provide graphs illustrating the effects on small cells after treatment with lipid-conjugated Gfap blunt-ended or tetracyclic oligonucleotides as assessed in FIGS. 8A to 8F and 9A to 9F . Percentage (%) of mouse Gfap mRNA remaining in the following regions of the mouse: lumbar spinal cord ( Figure 10A ), medulla oblongata ( Figure 10B ), cerebellum ( Figure 10C ), hypothalamus ( Figure 10D ), hippocampus ( Figure 10E ) , and frontal cortex ( Figure 10F ). Experiment 1 represents the oligonucleotides evaluated in Figures 8A to 8F . Experiment 2 represents the tetracyclic oligonucleotides evaluated in Figures 9A to 9F . Experiment 3 represents the blunt-ended oligonucleotides evaluated in Figures 9A - 9F . [ Figure 11A ] to [ Figure 11B ] provide graphs illustrating the concentration response of C16- bound Gfap blunt-ended oligonucleotides. Percentage (%) of murine Gfap mRNA remaining in the CNS of mice after treatment with C16-conjugated Gfap blunt-ended oligonucleotides. Mice were treated with 3, 10, 30, 100, or 300 µg of the indicated oligonucleotides in artificial cerebrospinal fluid (aCSF) via intrathecal injection into the lumbar spine. On days seven (7) ( Figure 11A ) and 28 ( Figure 11B ) after intrathecal injection, Gfap mRNA levels were normalized to ribosomal protein L23 (RPL23) mRNA and determined relative to control mice treated with aCSF Overall performance and ED ₅₀ between tissue types.

TW202335674A_111142234_SEQL.xml

Claims (101)

A double-stranded oligonucleotide comprising an antisense strand of 15 to 30 nucleotides long and a sense strand of 15 to 50 nucleotides long, wherein the antisense strand and the sense strand form 15 to 30 bases For a duplex region, wherein the antisense strand includes a region complementary to a stellate cell mRNA target sequence, and wherein the sense strand includes at least one lipid moiety that binds to a nucleotide of the sense strand. The oligonucleotide of claim 1, wherein the lipid part is selected from The oligonucleotide of claim 1, wherein the lipid part is a hydrocarbon chain. The oligonucleotide of claim 3, wherein the hydrocarbon chain is a C8 to C30 hydrocarbon chain. The oligonucleotide of claim 3 or 4, wherein the hydrocarbon chain is a C16 hydrocarbon chain. The oligonucleotide of claim 5, wherein the C16 hydrocarbon chain is composed of represented. The oligonucleotide of any one of claims 1 to 6, wherein the lipid moiety is combined with the 2' carbon of the ribose ring of the nucleotide. The oligonucleotide of any one of claims 1 to 7, wherein the oligonucleotide is blunt-ended. The oligonucleotide of claim 8, wherein the oligonucleotide is blunt-ended at the 3' end of the oligonucleotide. The oligonucleotide of any one of claims 1 to 7, wherein the oligonucleotide includes a blunt end. The oligonucleotide of claim 10, wherein the blunt end includes the 3' end of the sense strand. The oligonucleotide of any one of claims 8 to 11, wherein the sense strand is 20 to 22 nucleotides. The oligonucleotide of claim 12, wherein the at least one lipid moiety binds to a nucleotide at position 1, position 4, position 8, position 12, position 13, position 18, or position 20 of the sense strand , where the positions are numbered from 5' to 3'. The oligonucleotide of claim 12, wherein the stellate cell mRNA target is expressed in the spinal cord, and wherein the at least one lipid moiety is associated with position 1, position 4, position 8, position 12, and position of the sense strand 13. The nucleotide at position 18, or position 20 is bound, and the positions are numbered from 5' to 3'. The oligonucleotide of claim 12, wherein the stellate cell mRNA target is expressed in the medulla oblongata, and wherein the at least one lipid moiety is associated with position 1, position 4, position 8, position 12, and position of the sense strand 13. The nucleotide at position 18, or position 20 is bound, and the positions are numbered from 5' to 3'. The oligonucleotide of claim 12, wherein the stellate cell mRNA target is expressed in the cerebellum, and wherein the at least one lipid moiety is associated with position 4, position 12, position 13, position 18, or The nucleotide at position 20 binds, and the positions are numbered from 5' to 3'. The oligonucleotide of claim 12, wherein the stellate cell mRNA target is expressed in the hypothalamus, and wherein the at least one lipid moiety is associated with position 1, position 4, position 12, and position 13 of the sense strand , position 18, or position 20, where the positions are numbered from 5' to 3'. The oligonucleotide of claim 12, wherein the stellate cell mRNA target is expressed in the frontal cortex, wherein the at least one lipid moiety binds to the nucleotide at position 4 of the sense strand, and wherein Positions are numbered from 5' to 3'. The oligonucleotide of any one of claims 1 to 7, wherein the sense strand includes a backbone-loop at the 3' end, wherein the backbone-loop includes the formula: 5'-S1-L-S2 The nucleotide sequence represented by -3', wherein S1 is complementary to S2, and wherein L forms a loop between S1 and S2. The oligonucleotide of any one of claims 1 to 7 and 19, wherein the sense strand is 36 to 38 nucleotides. The oligonucleotide of claim 20, wherein the stellate cell mRNA target is expressed in the spinal cord, and wherein the at least one lipid moiety is associated with position 1, position 4, position 8, position 12, and position of the sense strand 13. Nucleotide binding at position 18, position 20, position 23, position 28, position 29, or position 30, and the positions are numbered from 5' to 3'. The oligonucleotide of claim 20, wherein the stellate cell mRNA target is expressed in the medulla oblongata, and wherein the at least one lipid moiety is associated with position 1, position 4, position 18, position 20, and position of the sense strand 23. Nucleotide binding at position 28, position 29, or position 30, and wherein the positions are numbered from 5' to 3'. The oligonucleotide of claim 20, wherein the stellate cell mRNA target is expressed in the cerebellum, and wherein the at least one lipid moiety is associated with position 1, position 4, position 23, position 28, or The nucleotide at position 29 binds, and the positions are numbered from 5' to 3'. The oligonucleotide of claim 20, wherein the stellate cell mRNA target is expressed in the hypothalamus, and wherein the at least one lipid moiety is associated with position 1, position 4, position 12, and position 13 of the sense strand , a nucleotide binding at position 18, position 20, position 23, position 28, position 29, or position 30, and wherein the positions are numbered from 5' to 3'. The oligonucleotide of claim 20, wherein the stellate cell mRNA target is expressed in the frontal cortex, wherein the at least one lipid moiety binds to the nucleotide at position 23 of the sense strand, and wherein Positions are numbered from 5' to 3'. The oligonucleotide of any one of claims 1 to 25, wherein the antisense strand is 22 to 24 nucleotides. The oligonucleotide of any one of claims 1 to 26, wherein the double-stranded region is 20 to 22 base pairs. The oligonucleotide of any one of claims 1 to 27, wherein the antisense strand comprises 1 to 4 nucleotide overhangs at the 3' end. The oligonucleotide of claim 28, wherein the overhang includes a purine nucleotide. The oligonucleotide of claim 28 or 29, wherein the overhang sequence is 2 nucleotides long. The oligonucleotide of claim 30, wherein the overhang is selected from AA, GG, AG, and GA. The oligonucleotide of claim 31, wherein the overhang is GG or AA. The oligonucleotide of claim 31, wherein the overhang is GG. The oligonucleotide of any one of claims 1 to 33, wherein the complementary region is complementary to at least 15 consecutive nucleotides of the stellate cell mRNA target sequence. The oligonucleotide of any one of claims 1 to 34, wherein the complementary region is complementary to 19 consecutive nucleotides of the stellate cell mRNA target sequence. The oligonucleotide of any one of claims 1 to 35, wherein the complementary region is completely complementary to the stellate cell mRNA target sequence. The oligonucleotide of any one of claims 1 to 35, wherein the complementary region is partially complementary to the stellate cell mRNA target sequence. The oligonucleotide of claim 37, wherein the complementary region contains no more than four mismatches with the stellate cell mRNA target sequence. The oligonucleotide of any one of claims 1 to 38, wherein the oligonucleotide comprises at least one modified nucleotide. The oligonucleotide of claim 39, wherein the modified nucleotide comprises a 2'-modification. The oligonucleotide of claim 40, wherein in addition to the nucleotide of the sense strand bound to the at least one lipid moiety, each of the nucleotides of the sense strand and the antisense strand comprise 2 '-modification. The oligonucleotide of claims 40 to 41, wherein the 2'-modification is selected from the following modifications: 2'-aminoethyl, 2'-fluoro, 2'-O-methyl, 2'- O-methoxyethyl, and 2'-deoxy-2'-fluoro-β-d-arabinonucleic acid (2'-deoxy-2'-fluoro-β-d-arabinonucleic acid). For example, the oligonucleotide of any one of claims 40 to 42, wherein about 10 to 20%, 10%, 11%, 12%, 13%, 14%, 15% of the nucleotides of the sense strand , 16%, 17%, 18%, 19%, or 20% contain 2'-fluorine modification. For example, the oligonucleotide of any one of claims 40 to 43, wherein about 25 to 35%, 25%, 26%, 27%, 28%, 29%, 30% of the nucleotides of the antisense strand %, 31%, 32%, 33%, 34%, or 35% contain a 2'-fluoro modification. Such as the oligonucleotide of any one of claims 40 to 44, wherein about 25 to 35%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, or 35% contained 2'-fluoro modifications. The oligonucleotide of any one of claims 40 to 45, wherein the sense strand includes 20 nucleotides and is from 5' to 3' at positions 1 to 20, wherein each of positions 8 to 11 includes 2' -Fluorine modification. The oligonucleotide of any one of claims 40 to 45, wherein the sense strand includes 20 nucleotides and is from 5' to 3' at positions 1 to 20, wherein each of positions 9 to 11 includes 2' -Fluorine modification. The oligonucleotide of any one of claims 40 to 45, wherein the sense strand comprises 36 nucleotides and is from 5' to 3' at positions 1 to 36, wherein each of positions 8 to 11 includes 2' -Fluorine modification. The oligonucleotide of any one of claims 40 to 45, wherein the sense strand includes 36 nucleotides and is from 5' to 3' at positions 1 to 36, wherein each of positions 9 to 11 includes 2' -Fluorine modification. The oligonucleotide of any one of claims 40 to 49, wherein the antisense strand includes 22 nucleotides and positions 1 to 22 from 5' to 3', and wherein positions 2, 3, 4, 5 Each of , 7, 10, and 14 contains a 2'-fluorine modification. The oligonucleotide of any one of claims 41 to 50, wherein in addition to the nucleotide of the sense strand bound to the at least one lipid moiety, the remaining nucleotides comprise 2'-O- Methyl modification. The oligonucleotide of any one of the preceding claims, wherein the oligonucleotide comprises at least one modified inter-nucleotide linkage. The oligonucleotide of claim 52, wherein the at least one modified internucleotide linkage is linked to a phosphorothioate linkage. The oligonucleotide of claim 53, wherein the antisense strand comprises a phosphorothioate linkage (i) between positions 1 and 2, and between positions 2 and 3; or (ii) at position Between 1 and 2, between positions 2 and 3, and between positions 3 and 4, where positions are numbered 1 to 4 from 5' to 3'. The oligonucleotide of claim 53 or 54, wherein the antisense strand is 22 nucleotides long, and wherein the antisense strand contains a phosphorothioate between positions 20 and 21 and between positions 21 and 22 Ester linkage where positions are numbered 1 to 22 from 5' to 3'. The oligonucleotide of any one of claims 53 to 55, wherein the sense strand comprises a phosphorothioate linkage between positions 1 and 2, wherein positions are numbered 1 to 2 from 5' to 3'. The oligonucleotide of any one of claims 53 to 56, wherein the sense strand is 20 nucleotides long, and wherein the sense strand is comprised between positions 18 and 19, and between positions 19 and 20 Phosphorothioate linkage where positions are numbered 1 to 22 from 5' to 3'. The oligonucleotide of any one of claims 1 to 57, wherein the antisense strand comprises a phosphorylated nucleotide at the 5' end, wherein the phosphorylated nucleotide is selected from uridine and adenosine. The oligonucleotide of claim 58, wherein the phosphorylated nucleotide is uridine. The oligonucleotide of any one of the preceding claims, wherein the 4'-carbon of the sugar of the 5'-nucleotide of the antisense strand comprises a phosphate analog. The oligonucleotide of claim 60, wherein the phosphate analog is oxymethylphosphonate, vinylphosphonate or malonylphosphonate. The oligonucleotide of any one of claims 1 to 61, wherein the complementary region is completely complementary to the stellate cell mRNA target sequence at nucleotide positions 2 to 8 of the antisense strand, wherein the nucleoside Acid positions are numbered from 5' to 3'. The oligonucleotide of any one of claims 1 to 61, wherein the complementary region is completely complementary to the stellate cell mRNA target sequence at nucleotide positions 2 to 11 of the antisense strand, wherein the nucleoside Acid positions are numbered from 5' to 3'. The oligonucleotide of any one of claims 1 to 63, wherein the oligonucleotide is a Dicer substrate. The oligonucleotide of any one of claims 1 to 63, wherein the oligonucleotide is a Dicer substrate and produces a double-stranded nucleic acid of 19 to 21 nucleotides in length after processing by the endogenous Dicer, It reduces stellate cell mRNA expression in mammalian cells. The oligonucleotide of any one of claims 1 to 65, wherein the stellate cell mRNA target sequence is located in the region of the central nervous system (CNS). The oligonucleotide of claim 66, wherein the region of the CNS is selected from the group consisting of the spinal cord, lumbar spinal cord, thoracic spinal cord, cervical spinal cord, medulla oblongata, hippocampus, cerebellum, hypothalamus, frontal cortex, and combinations thereof. The oligonucleotide of any one of claims 1 to 67, wherein the oligonucleotide reduces the expression of target mRNA in stellate cells or stellate cell populations in vitro and/or in vivo. A pharmaceutical composition comprising the oligonucleotide of any one of claims 1 to 68, and a pharmaceutically acceptable carrier, delivery agent or excipient. A method for treating an individual suffering from a disease, disorder or condition associated with expression of stellate cell mRNA, the method comprising administering to the individual a therapeutically effective amount of an oligonucleotide according to any one of claims 1 to 68 or a pharmaceutical composition as claimed in claim 69, thereby treating the individual. A method of delivering an oligonucleotide to a stellate cell or a population of stellate cells in an individual, the method comprising administering a pharmaceutical composition according to claim 69 to the individual. The method of claim 71, wherein the stellate cell or stellate cell population is located in a region of the CNS. The method of claim 72, wherein the region of the CNS is selected from the group consisting of the spinal cord, lumbar spinal cord, thoracic spinal cord, cervical spinal cord, medulla oblongata, hippocampus, cerebellum, hypothalamus, frontal cortex, and combinations thereof. A method for reducing the expression of stellate cell mRNA in a cell, cell population, or individual, the method comprising the following steps: i. Bringing the cell or cell population into contact with the oligonucleotide of any one of claims 1 to 68, or the pharmaceutical composition of claim 69, optionally wherein the cell or cell population is a stellate cell or a stellate cell population; or ii. Administer the oligonucleotide according to any one of claims 1 to 68, or the pharmaceutical composition according to claim 69 to the individual. The method of claim 74, wherein reducing the expression of the stellate cell mRNA comprises reducing the amount or level of mRNA, the amount or level of protein, or both. The method of claim 74 or 75, wherein the individual suffers from a disease, disorder or condition associated with expression of the stellate cell mRNA. The method of any one of claims 74 to 76, wherein the cell or cell population is located in a region of the CNS. The method of claim 77, wherein the region of the CNS is selected from the group consisting of the spinal cord, lumbar spinal cord, thoracic spinal cord, cervical spinal cord, medulla oblongata, hippocampus, cerebellum, hypothalamus, frontal cortex, and combinations thereof. The method of any one of claims 70 to 78, wherein the administration is intrathecally. A method of reducing the expression of target mRNA expressed in stellate cells in the tissue of the CNS of an individual, the method comprising administering to the individual a double-stranded oligonucleotide, the double-stranded oligonucleotide comprising 15 to 30 An antisense strand of 15 to 50 nucleotides long and a sense strand of 15 to 50 nucleotides long, wherein the antisense strand and the sense strand form a double-stranded region of 15 to 30 base pairs, wherein the antisense strand includes A region complementary to the target sequence in the target mRNA, and wherein the sense strand includes at least one lipid moiety that binds to a nucleotide of the sense strand. The method of claim 80, wherein the lipid moiety is a C16 hydrocarbon. The method of claims 80 to 81, wherein the oligonucleotide is blunt-ended at the 3' end of the oligonucleotide. The method of claim 82, wherein the sense strand is 22 to 24 nucleotides. The method of claim 83, wherein the tissue is the spinal cord, and wherein the at least one lipid moiety is associated with position 1, position 4, position 8, position 12, position 13, position 18, or position 20 of the sciatic strand. Nucleotides are bound, and positions therein are numbered from 5' to 3'. The method of claim 83, wherein the tissue is the medulla oblongata, and wherein the at least one lipid moiety is associated with position 1, position 4, position 8, position 12, position 13, position 18, or position 20 of the justice strand. Nucleotides are bound, and positions therein are numbered from 5' to 3'. The method of claim 83, wherein the tissue is the cerebellum, and wherein the at least one lipid moiety binds to a nucleotide at position 4, position 12, position 13, position 18, or position 20 of the sense strand, and The positions are numbered from 5' to 3'. The method of claim 83, wherein the tissue is the hypothalamus, and wherein the at least one lipid moiety is associated with the nucleus at position 1, position 4, position 12, position 13, position 18, or position 20 of the justice strand The nucleotides are bound and the positions are numbered from 5' to 3'. The method of claim 83, wherein the tissue is the frontal cortex, wherein the at least one lipid moiety binds to a nucleotide at position 4 of the sense strand, and wherein the positions are numbered from 5' to 3'. The method of any one of claims 80 to 81, wherein the justice strand includes a backbone-loop at the 3' end, wherein the backbone-loop includes the formula: 5'-S1-L-S2-3' Represented is a nucleotide sequence in which S1 is complementary to S2 and in which L forms a loop between S1 and S2. The method of claim 89, wherein the sense strand is 36 to 38 nucleotides. The method of claim 90, wherein the tissue is the spinal cord, and wherein the at least one lipid moiety is associated with position 1, position 4, position 8, position 12, position 13, position 18, position 20, position 23 of the sciatic strand , the nucleotide at position 28, position 29, or position 30 binds, and the positions are numbered from 5' to 3'. The method of claim 90, wherein the tissue is the medulla oblongata, and wherein the at least one lipid moiety is associated with position 1, position 4, position 18, position 20, position 23, position 28, position 29, or position of the justice strand 30 nucleotides are bound, and the positions are numbered from 5' to 3'. The method of claim 90, wherein the tissue is the cerebellum, wherein the at least one lipid moiety binds to a nucleotide at position 1, position 4, position 23, position 28, or position 29 of the sense strand, and The positions are numbered from 5' to 3'. The method of claim 90, wherein the tissue is the hypothalamus, and wherein the at least one lipid moiety is associated with positions 1, 4, 12, 13, 18, 20, 23, The nucleotide at position 28, position 29, or position 30 binds, and the positions are numbered from 5' to 3'. The method of claim 90, wherein the tissue is the frontal cortex, wherein the at least one lipid moiety binds to nucleotide at position 23 of the sense strand, and wherein the positions are numbered from 5' to 3'. The method of any one of claims 70 to 95, wherein a single dose of the oligonucleotide or pharmaceutical composition reduces the expression of the stellate cell mRNA for at least 4 weeks, at least 8 weeks, at least 12 weeks, at least 23 weeks weeks, at least 26 weeks, or at least 29 weeks. The method of any one of claims 70 to 95, wherein a single dose of the oligonucleotide or pharmaceutical composition reduces the expression of the stellate cell mRNA for up to one year. A kit comprising the oligonucleotide of any one of claims 1 to 68, a pharmaceutically acceptable carrier if necessary, and a drug instruction sheet, the drug instruction sheet containing a medicine for treating patients with a star Instructions for administration to individuals with diseases, disorders, or conditions associated with expression of cellular mRNA. For example, the kit of claim 98, wherein the drug package insert includes instructions for intrathecal administration. Use of an oligonucleotide according to any one of claims 1 to 68 or a pharmaceutical composition according to claim 69 in the manufacture of a medicament for the treatment of diseases, disorders or conditions associated with the expression of stellate cell mRNA. The oligonucleotide of any one of claims 1 to 68 or the pharmaceutical composition of claim 69 is for use, or is applicable, in the treatment of diseases, disorders or conditions associated with the expression of stellate cell mRNA.