WO2011118819A1 - Thérapie génique pour une tumeur du tissue hématopoïétique par vecteur viral à prolifération régulée portant un promoteur de la survivine - Google Patents

Thérapie génique pour une tumeur du tissue hématopoïétique par vecteur viral à prolifération régulée portant un promoteur de la survivine Download PDF

Info

Publication number
WO2011118819A1
WO2011118819A1 PCT/JP2011/057477 JP2011057477W WO2011118819A1 WO 2011118819 A1 WO2011118819 A1 WO 2011118819A1 JP 2011057477 W JP2011057477 W JP 2011057477W WO 2011118819 A1 WO2011118819 A1 WO 2011118819A1
Authority
WO
WIPO (PCT)
Prior art keywords
promoter
hematopoietic tumor
gene
vector
survivin
Prior art date
Application number
PCT/JP2011/057477
Other languages
English (en)
Japanese (ja)
Inventor
紳介 鈴木
直道 有馬
健一郎 小戝
Original Assignee
Suzuki Shinsuke
Arima Naomichi
Kosai Ken-Ichiro
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Suzuki Shinsuke, Arima Naomichi, Kosai Ken-Ichiro filed Critical Suzuki Shinsuke
Publication of WO2011118819A1 publication Critical patent/WO2011118819A1/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/76—Viruses; Subviral particles; Bacteriophages
    • A61K35/761—Adenovirus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10332—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10341—Use of virus, viral particle or viral elements as a vector
    • C12N2710/10343—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2830/00—Vector systems having a special element relevant for transcription
    • C12N2830/008—Vector systems having a special element relevant for transcription cell type or tissue specific enhancer/promoter combination

Definitions

  • the present invention relates to a hematopoietic tumor therapeutic agent comprising a growth-controlling viral vector containing a Survivin promoter, or a hematopoietic tumor containing the vector, such as adult T cell leukemia (ATL) cells.
  • ATL adult T cell leukemia
  • the present invention also relates to a method for producing a hematopoietic tumor therapeutic agent, which comprises infecting a hematopoietic tumor, for example, an ATL cell, with a growth-controlling viral vector containing a Survivin promoter.
  • ATL also referred to as adult T-cell leukemia lymphoma (ATLL)
  • ATL is a blood cancer (hematopoietic tumor) caused by human T-cell leukemia virus type I (HTLV-1).
  • HTLV-1 human T-cell leukemia virus type I
  • Japan there are about 200,000 people infected with HTLV-1 mainly in South Kyushu, and more than 100 ATL patients occur every year. ATL still has no definitive treatment, and the development of new treatments is urgent.
  • CRA cancer-specific replicating adenovirus
  • CRA is a modification of the E1 gene region essential for the growth of adenovirus to differentiate the virus growth between cancer cells and normal cells.
  • Necessary for the growth of adenovirus in the E1 region A type that inhibits the inactivation of Rb and p53, which are essential for inducing a normal cellular environment, and inhibits their inactivation (ie, prevents viral growth in normal cells), and
  • E1 There is a type in which the endogenous promoter of a gene is replaced with a promoter of a gene that is highly expressed specifically in cancer, whereby the E1 gene is expressed specifically (a virus is propagated specifically in cancer).
  • CRA assumes local administration for solid cancer.
  • CRA's strategy for treating solid tumors is to inject CRA into the main cancer nodules, cause the main nodules to die, and then search for metastatic foci throughout the body to eradicate the cancer.
  • hematopoietic tumors, especially ATL usually have advanced stage lesions and target leukocyte floating cells, multiple lymph node lesions, and invasive lesions in multiple organs such as bone marrow, liver, spleen, and gastrointestinal tract. Therefore, a CRA administration method different from that for solid cancer is required.
  • An object of the present invention is to provide a novel dosage form of the vector that enables gene therapy of hematopoietic tumor using a growth-controlling viral vector, and a novel treatment of hematopoietic tumor using the dosage form To provide a strategy.
  • the present inventors focused on the fact that ATL cells highly express survivin, and using Survivin promoter-dependent CRA (Surv.CRA) originally developed by Obata et al., Surv for ATL treatment
  • Surv.CRA Survivin promoter-dependent CRA
  • Surv.CRA replication and infection of uninfected ATL cell lines spread from infected ATL cell lines.
  • Surv.CRA was a virus that showed sufficient infection efficiency and killing effect on ATL cells, and did not induce virus growth and cell lysis in normal lymphocytes, indicating that this virus vector has high ATL specificity. confirmed.
  • the present inventors have isolated a propagating viral vector in which a nucleic acid encoding a protein required for viral replication is under the control of a promoter of a gene that is specifically expressed in a hematopoietic tumor.
  • a propagating viral vector in which a nucleic acid encoding a protein required for viral replication is under the control of a promoter of a gene that is specifically expressed in a hematopoietic tumor.
  • Virus release occurs around the target leukopoietic lesions, multiple lymph node lesions, and infiltrating lesions in multiple organs such as bone marrow, liver, spleen, and gastrointestinal tract, effectively infecting these target tumor cells and killing them
  • the present invention has been found to be able to be induced.
  • the present invention provides: [1] The hematopoiesis comprising a proliferative virus vector containing a nucleic acid encoding a protein necessary for replication of at least one virus under the control of a promoter of a gene specifically expressed in hematopoietic tumor cells Therapeutic agents for organ tumors; [2] The agent according to [1] above, which is systemically administered in a form infected with hematopoietic tumor cells; [3] A hematopoietic tumor cell infected with a proliferative virus vector containing a nucleic acid encoding a protein necessary for replication of at least one virus under the control of a promoter of a gene specifically expressed in the hematopoietic tumor cell.
  • a therapeutic agent for the hematopoietic tumor comprising: [4] The agent according to any one of [1] to [3] above, wherein the viral vector is an adenoviral vector; [5] The agent according to any one of [1] to [4] above, wherein the promoter is a survivin promoter; [6] The above [5], wherein the Survivin promoter comprises at least the 1124 to 1278th nucleotide sequence in the nucleotide sequence shown in SEQ ID NO: 1, or the at least 1296 to 1468th nucleotide sequence in the nucleotide sequence shown in SEQ ID NO: 3.
  • the agent according to [7] The agent according to any one of [1] to [6] above, wherein the hematopoietic tumor is adult T-cell leukemia; [8] The agent according to any one of [1] to [7] above, wherein the vector further comprises a therapeutic gene for hematopoietic tumor; [9] A hematopoietic tumor cell infected with a proliferative virus vector containing a nucleic acid encoding a protein necessary for replication of at least one virus under the control of a promoter of a gene specifically expressed in the hematopoietic tumor cell.
  • a method of treating a hematopoietic tumor in said subject comprising systemic administration to the subject having a hematopoietic tumor; [10] The method according to [9] above, wherein the viral vector is an adenoviral vector; [11] The method according to [9] or [10] above, wherein the promoter is a survivin promoter; [12] The above [11], wherein the Survivin promoter comprises at least the 1124 to 1278th nucleotide sequence in the nucleotide sequence shown in SEQ ID NO: 1, or the at least 1296 to 1468th nucleotide sequence in the nucleotide sequence shown in SEQ ID NO: 3.
  • FIG. 2 shows the nucleotide sequence of the mouse (FIG. 2A) and human (FIG. 2B) survivin promoter and the consensus binding sequence of transcriptional regulators (boxed).
  • the arrow indicates the putative transcription start point, and the underlined ATG indicates the start codon.
  • the horizontal axis indicates the number of days after infection, and the vertical axis indicates the viability of Surv.CRA-infected cells (% relative to uninfected cells).
  • FIG. 4A shows ⁇ -galactosidase activity in a leukemia cell line infected with a non-proliferating adenovirus having a LacZ gene linked downstream of the survivin promoter.
  • FIG. 4B shows the relationship of IC50 values to Surv.CRA leukemia cell lines.
  • FIG. 1 It is a figure which shows the expansion of the virus replication in a Surv.CRA infection leukemia cell, and the infection to an uninfected leukemia cell.
  • the vertical axis shows the expression rate of EGFP in leukemia cell lines infected with Surv.CRA containing the EGFP gene.
  • Ad. ⁇ E1 shows the results of cells infected with non-proliferating adenovirus containing the EGFP gene and lacking the E1 region.
  • the propagating virus vector used in the present invention is a replica of at least one virus under the control of a promoter of a gene specifically expressed in hematopoietic tumor cells.
  • hematopoietic tumor blood cancer
  • hematopoietic tumor blood cancer
  • Leukemia is classified into acute leukemia and chronic leukemia according to the degree of progression, and depending on the type of cells proliferating, myeloid leukemia originating from myeloid cells and lymphocytic leukemia originating from lymphoid cells are categorized.
  • causes of hematopoietic tumors include radiation, chemical substances (eg, benzene, toluene, etc.), drugs (eg, anticancer agents, etc.), viruses (eg, HTLV-1, EB virus, etc.) and the like.
  • HTLV-1 is the cause of adult T-cell leukemia (/ lymphoma) (ATL / ATLL)
  • EB virus is the causative virus for Burkitt lymphoma (/ leukemia) (BL / BLL).
  • genes specifically expressed in hematopoietic tumor cells include survivin gene, WT1 gene, Aurora gene, FLT3 gene, BCR-ABL gene, JAK2 gene, c-MYC gene, BCL-1 gene, and BCL-2 gene. , BCL-6 gene, cyclin D1 gene, E2A gene, NOTCH1 gene and the like, but are not limited thereto.
  • survivin gene, WT1 gene, Aurora gene, FLT3 gene, BCL-2 gene, NOTCH1 gene and the like more preferably survivin gene.
  • “specific for hematopoietic tumor cells” is not limited to the case where no transcription activity is shown in normal cells, but the gene is also expressed in normal cells within the therapeutically acceptable range.
  • a survivin gene promoter is used as a promoter of a gene specifically expressed in hematopoietic tumor cells.
  • a promoter region can be isolated by the same technique, and a proliferative virus vector whose growth is controlled by the promoter can be constructed, and hematopoietic tumor cells that highly express the promoter in the virus can be constructed. It can be infected and administered to patients.
  • the promoters of mouse and human survivin genes have been isolated and their sequence information has been disclosed (eg Li, F. and Altieri, DC, Cancer Res., 59: 3143-3151, 1999; Li, F and Altieri, DC, Biochem. J., 344: 305-311, 1999).
  • the survivin promoter used in the vector of the present invention the human survivin gene or its ortholog gene in other mammals (eg, monkey, cow, horse, pig, dog, cat, sheep, goat, rabbit, mouse, rat, etc.) Promoters, preferably human or mouse derived survivin gene promoters, more preferably human survivin gene promoters.
  • a promoter may be used.
  • a vector containing a mouse survivin gene promoter can be used as a therapeutic vector for human hematopoietic tumors.
  • the survivin promoter nucleotide sequence length is specific to the target hematopoietic tumor cell (eg, ATL cell) and transcribes the downstream linked gene to the extent that it exhibits sufficient therapeutic activity against the hematopoietic tumor. There is no particular limitation as long as it can be activated.
  • the nucleotide sequence at positions ⁇ 173 to ⁇ 19 the nucleotide sequence from 1124 to 1278 in the nucleotide sequence shown in SEQ ID NO: 1, where the translation start point is +1
  • the human survivin gene promoter If the nucleotide sequence at positions -173 to -1 (the nucleotide sequence from 1296 to 1468 in the nucleotide sequence shown in SEQ ID NO: 2) is included with the translation start point as +1, the desired specificity and transcription activity can be obtained. (See, for example, FIG. 3).
  • the survivin promoter used in the vector of the present invention has at least the 1124 to 1278th nucleotide sequence in the nucleotide sequence shown in SEQ ID NO: 1, or the at least 1296 to 1468th nucleotide in the nucleotide sequence shown in SEQ ID NO: 2.
  • the upper limit of the nucleotide sequence length of the survivin promoter is not particularly limited, but if the length of the 5 'upstream region is too large, it may adversely affect the transcription activity and specificity of the promoter.
  • the desired specificity and transcriptional activity can be obtained as long as the nucleotide sequence is about -6000 to -1 with the translation start point being +1.
  • the 5 ′ end of the promoter is It is downstream from the -3000 position, more preferably downstream from the -1500 position.
  • vectors with reporter genes linked downstream of promoters of various lengths are prepared and introduced into target hematopoietic tumor cells. By evaluating the promoter activity using expression as an index, a suitable range of the sequence length of the promoter can be determined.
  • a survivin promoter is also a nucleic acid that can hybridize under stringent conditions with a natural mammal-derived (ie, wild-type) survivin promoter, and that has substantially the same characteristics as the natural promoter. Include. “Substantially the same property” means a property that drives gene expression specific to a target hematopoietic tumor cell, and the degree of transcriptional activity is preferably equivalent (eg, about 0.5 to about 2 times). However, the quantitative factors may be different as long as they can drive gene expression to the extent that sufficient therapeutic activity can be exerted on the target hematopoietic tumor.
  • a nucleic acid that can hybridize under stringent conditions with the complementary strand sequence of the nucleotide sequence 1124 to 1278 in the nucleotide sequence shown in SEQ ID NO: 1 is also a human survivin promoter.
  • nucleic acid for example, about 80% or more, preferably about 90% or more, more preferably about 95% or more, particularly preferably about 97% or more, and most preferably, the partial nucleotide sequence of SEQ ID NO: 1 or 2 Include nucleic acids containing nucleotide sequences having about 98% or more homology.
  • Hybridization can be performed according to a method known per se or a method analogous thereto, for example, the method described in Molecular Cloning, 2nd ed. (J. Sambrook et al., Cold Spring Harbor Lab. Press, 1989). When a commercially available library is used, hybridization can be performed according to the method described in the attached instruction manual. Hybridization can be preferably performed according to highly stringent conditions.
  • Stringent conditions include: (1) Use low ionic strength and high temperature for washing, for example, 0.015 M sodium chloride / 0.0015 M sodium citrate / 0.1% sodium dodecyl sulfate at 50 ° C, and (2) formamide 50% (v / v) with a denaturant, for example, 0.1% bovine serum albumin / 0.1% Ficoll / 0.1% polyvinylpyrrolidone / 750 mM sodium chloride, 50 mM sodium phosphate buffer (pH 6.5) containing 75 mM sodium citrate.
  • v) Reaction conditions characterized by using formamide at 42 ° C are exemplified.
  • stringent conditions include 50% formamide, 5xSSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5x Denhart solution, sonicated sperm DNA (50 mg / ml), 0.1% SDS, and 10% dextran sulfate at 42 ° C, washed with 0.2xSSC and 50% formaldehyde at 55 ° C, followed by 0.1xSSC containing EDTA at 55 ° C High stringency washing may be performed.
  • a person skilled in the art can easily achieve a desired stringency by appropriately adjusting the temperature during the hybridization reaction and / or washing, the ionic strength of the buffer, and the like according to factors such as the probe length.
  • Survivin promoter is known from genomic DNA extracted from cells and tissues derived from humans or other mammals (eg monkeys, cows, horses, pigs, dogs, cats, sheep, goats, rabbits, mice, rats, etc.) Survivin gene promoter sequences (eg, Li, F. and Altieri, DC, Cancer Res., 59: 3143-3151, 1999; Li, F. and Altieri, DC, Biochem.
  • the genomic DNA containing the promoter region is cloned using a nucleic acid consisting of a DNA fragment as a probe, cleaved into a DNA fragment containing the desired partial promoter sequence using a DNA-degrading enzyme such as an appropriate restriction enzyme, and separated by gel electrophoresis. Thereafter, it can be prepared by collecting the desired band and purifying the DNA.
  • the survivin promoter partial sequence may be amplified and isolated by PCR using the above crude cell extract or genomic DNA isolated therefrom as a template and a primer synthesized based on a known survivin gene promoter sequence. You can also.
  • the survivin promoter is based on a known survivin gene promoter sequence (for example, the nucleotide sequence shown in SEQ ID NO: 1 or 2), and a nucleic acid containing all or part of the nucleotide sequence is converted into a commercially available DNA / RNA automatic It can also be obtained by chemical synthesis using a synthesizer.
  • the vector of the present invention is a propagation type (restriction propagation type) viral vector. That is, the target hematopoietic tumor cells are characterized by specific growth (dominantly over normal cells).
  • the vector of the present invention can be constructed by placing a nucleic acid encoding a protein required for replication of at least one virus under the control of a survivin promoter.
  • proteins necessary for viral replication refers to viral proteins required for transcription of various viral genes (for example, adenovirus E1A, E1B, E2 and E4 which are early genes before transcription of viral structural proteins).
  • E1A is the first protein that is transcribed and translated after infection, and is essential for the initiation of transcription of various viral proteins that are followed by transcription of this E1A protein), or the structure of the virus A gene (for example, L1, L2, L3, L4, L5, etc., which is a late gene in adenovirus) means any gene that is essential for the virus to self-replicate, and varies depending on the virus type used.
  • adeno-associated virus For example, Rep78 and Rep68 under the control of the p5 promoter, Rep52 and Rep40 under the control of the p19 promoter, such as Herpes simplex virus, early gene products such as ICP0, ICP4, ICP22, ICP27, thymidine kinase, etc.
  • the p19 promoter such as Herpes simplex virus, early gene products such as ICP0, ICP4, ICP22, ICP27, thymidine kinase, etc.
  • viruses N protein, P protein, L protein, etc.
  • Such a propagating viral vector can be obtained by replacing the endogenous promoter of a nucleic acid encoding a protein necessary for at least one viral replication with a survivin promoter.
  • the vector of the present invention is an adenoviral vector, preferably a nucleic acid encoding E1A and / or E1B, more preferably a nucleic acid encoding at least E1A is placed under the control of a survivin promoter.
  • the proliferative virus vector introduced into the cell cannot grow in an environment where the survivin promoter is not activated (for example, normal cells), the cell is not damaged.
  • the vector of the present invention enters the environment in which the survivin promoter is activated (that is, the target hematopoietic tumor cell)
  • the virus grows there, and the cell is damaged by the cytotoxicity of the viral protein.
  • the virus released from the lysed cells infects surrounding hematopoietic tumor cells into which no vector has been introduced, and further hematopoietic tumor cells throughout the body through blood circulation, and the same steps are repeated.
  • the vector of the present invention can be finally introduced into all hematopoietic tumor cells.
  • Proteins required for virus replication are essential for inducing the cellular environment necessary for virus growth in normal cells, but lacking regions not required for virus growth in target hematopoietic tumor cells. There may be.
  • Rb and p53 in order to propagate virus in normal cells, it is necessary to inactivate Rb and p53 in order to turn the cell cycle, but hematopoietic tumor cells are already in a state where the cell cycle is turning.
  • the proliferative adenoviral vector of the present invention can allow hematopoietic tumor cell-specific viral growth by deleting the E1A24KDa, E1B55KDa, or E1B19KDa region.
  • the vector of the present invention contains a nucleic acid encoding a protein necessary for replication of two or more viruses
  • viral propagation is survivin if at least one of these nucleic acids is under the control of a survivin promoter. Since it is limited to the environment in which the promoter is activated, the nucleic acid encoding a protein required for replication of other viruses may be placed under the control of any promoter.
  • a cytomegalovirus (CMV) -derived promoter eg, CMV immediate early promoter
  • a human immunodeficiency virus (HIV) -derived promoter eg, HIV LTR
  • a rous sarcoma virus (RSV) -derived promoter eg, RSV LTR
  • Promoter derived from mouse mammary tumor virus (MMTV) eg, MMTV LTR
  • promoter derived from Moloney murine leukemia virus (MoMLV) eg, MoMLV LTR
  • promoter derived from herpes simplex virus (HSV) eg, HSV thymidine kinase (TK) promoter
  • HSV40-derived promoter eg, SV40 early promoter
  • AAV adeno-associated virus
  • AdV adenovirus
  • a metallothionein-1 gene promoter can be used as the inducible promoter.
  • administration of a heavy metal such as gold, zinc, or cadmium, a steroid such as dexamethasone, an alkylating agent, a chelating agent, or a cytokine at any desired time In the target hematopoietic tumor cell can be specifically induced viral protein expression.
  • the promoters used may be the same or different. For example, a human survivin promoter and a mouse survivin promoter can be used together in one vector.
  • nucleic acid encoding a protein required for virus replication controlled by a promoter other than the survivin promoter it is essential for inducing the cellular environment necessary for virus growth in normal cells, but the target hematopoietic tumor It is also possible to use a nucleic acid encoding the above mutant virus protein in which a region not necessary for virus growth in cells is deleted.
  • the vector of the present invention can further contain a therapeutic gene for hematopoietic tumor in addition to a nucleic acid encoding a protein necessary for virus replication.
  • therapeutic gene encodes any protein (or RNA) as long as the gene, when transcribed (and translated), directly or indirectly has a therapeutic effect on the target hematopoietic tumor.
  • tumor suppressor genes p53, p21, etc.
  • cytokine genes GM-CSF, IL-2, IL-4, IFN, etc.
  • apoptosis-inducing genes Fas, etc.
  • ion channels Sodium channel, etc.
  • Genes that encode constituent proteins, protein genes that can damage cells by converting prodrugs into poisons suicide genes
  • suicide genes HSV-thymidine kinase, cytosine deaminase, etc.
  • anti-cancer genes Sense nucleic acid (antisense nucleic acid for TGF- ⁇ , antisense nucleic acid for survivin, etc.), angiogenesis inhibition Gene (platelet IV factor, angiostatin, endostatin, soluble VEGF receptor, etc.) and the like.
  • the vector of the present invention may further comprise a visualization gene (eg, GFP, EGFP, RFP, luciferase, LacZ gene, etc.) for identifying hematopoietic tumor cells infected with the vector instead of or in addition to the therapeutic gene.
  • a visualization gene eg, GFP, EGFP, RFP, luciferase, LacZ gene, etc.
  • the therapeutic gene may be placed under the control of any promoter.
  • a constitutive promoter such as the above-mentioned viral promoter or a gene promoter of a mammalian constituent protein, a promoter of a gene that can be specifically expressed in a target hematopoietic tumor cell, an inducible promoter, and the like can be similarly used. it can.
  • the therapeutic gene can also be placed under the control of the survivin promoter.
  • Nucleic acids encoding therapeutic proteins necessary for viral replication and therapeutic genes can be isolated as cDNA from cells / tissues that produce them by methods known per se, downstream of the survivin promoter or any other promoter. Can be functionally linked.
  • An expression cassette containing a nucleic acid encoding a protein required for viral replication or a therapeutic gene under the control of a survivin promoter or any other promoter is preferably a suitable polyadenylation sequence downstream of the nucleic acid or gene. including.
  • the vector of the present invention imparts resistance to a replication origin for autonomous amplification in host cells and a selection marker gene for selection of transformed cells (tetracycline, ampicillin, kanamycin, hygromycin, phosphinothricin, etc.) Or a gene that complements an auxotrophic mutation).
  • a selection marker gene for selection of transformed cells (tetracycline, ampicillin, kanamycin, hygromycin, phosphinothricin, etc.) Or a gene that complements an auxotrophic mutation).
  • the vector of the present invention may be any virus-derived vector as long as a restricted propagation vector can be constructed, and examples thereof include adenovirus, adeno-associated virus, herpes virus, Sendai virus, and the like, preferably Is an adenovirus vector.
  • a multifactor cancer-specific growth-regulated recombinant adenovirus system (m-CRA; JP 2005-046101 and WO 2005/012536) developed by Kominato et al.
  • a survivin promoter can be used.
  • An example of a plasmid vector suitably used for construction of m-CRA is presented in FIG.
  • survivin promoter is used as promoter A and / or promoter B of plasmid vector P1
  • survivin promoter or another target hematopoietic organ is used as promoter C (controls the expression of therapeutic gene) of plasmid vector P2.
  • Any other promoter such as a tumor cell specific promoter or a constitutive promoter can be used.
  • the target hematopoietic tumor cell-specific promoter include promoters of the above various genes that are specifically expressed in hematopoietic tumor cells.
  • the E1A gene (which may lack the 24 KDa region) operably linked to the survivin promoter, and a constitutive promoter (such as a CMV promoter)
  • a constitutive promoter such as a CMV promoter
  • a reporter gene (as a model system for therapeutic genes) operably linked to a constitutive promoter (such as a CMV promoter)
  • a plasmid vector P2 is provided, as well as a backbone plasmid P3 containing an adenoviral genome (which may have a target cell specific mutation in the fiber gene) lacking the E1 region.
  • a CRA vector can be prepared by transfection into a cell line (eg, 293 cells) that complements E1.
  • the growth of the vector is controlled by two factors, the promoter controlling the expression of E1A (survivin promoter) and the E1B gene (E1B ⁇ 55K), but the expression of the E1A gene and E1B is controlled.
  • the promoter controlling the expression of E1A (survivin promoter)
  • the E1B gene E1B ⁇ 55K
  • the expression of the E1A gene and E1B is controlled.
  • the vector of the present invention is a restricted-proliferation viral vector in which a nucleic acid encoding a protein required for at least one viral replication is under the control of a promoter of a gene (eg, survivin) that is specifically expressed in a hematopoietic tumor. Therefore, the virus cannot grow in cells in which the promoter does not exert transcriptional activity. Therefore, the vector of the present invention comprises introducing a protein necessary for replication of a virus encoded by a nucleic acid under the control of a promoter of a gene specifically expressed in a hematopoietic tumor into a cell that is complementary, and culturing the cell. Can be amplified.
  • a promoter of a gene eg, survivin
  • the 293 cell line in which the E1A and E1B genes are incorporated can be used as a complementary cell line.
  • the amplified virus can be recovered from the culture supernatant of the cells according to a conventional method.
  • the vector of the present invention is administered systemically as it is, for example, in the case of an adenovirus vector, the administered vector is abundant in the liver and spleen, and it is known that it corresponds to 20 times the amount of CRA in tumor cells. Even if this method is applied to hematopoietic tumors, there is a high possibility that sufficient infection efficiency cannot be obtained for tumor cells in peripheral blood.
  • the above vector is infected with the isolated hematopoietic tumor cell, and the infected hematopoietic tumor cell is systemically administered to a patient suffering from the hematopoietic tumor, thereby being transferred into the blood.
  • the present invention provides a therapeutic agent for hematopoietic tumors, comprising hematopoietic tumor cells infected with the above-mentioned proliferative virus vector.
  • Hematopoietic tumor patients who can be treated with a preparation containing hematopoietic tumor cells infected with the vector of the present invention can exhibit transcriptional activity at a level at which the hematopoietic tumor-specific promoter mounted on the vector is therapeutically effective.
  • Any animal having a hematopoietic tumor is not particularly limited.
  • patients with acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, multiple myeloma include adult T-cell leukemia / lymphoma.
  • the hematopoietic tumor cells to be infected with the vector of the present invention are administered to an animal (preferably human) to be treated, it is particularly preferable that they are hematopoietic tumor cells collected from the animal individual, When it is difficult to collect tumor cells from the treatment subject itself, tumor cells can be collected from other individuals having the same or substantially the same type of MHC.
  • the “substantially the same” type of MHC means that the type of MHC matches to such an extent that the transplanted cells can be engrafted when the tumor cells are transplanted into an animal to be treated by using an immunosuppressant or the like. It means that For example, in the case of a human, for example, main HLA (for example, three loci of HLA-A, HLA-B and HLA-DR) are the same.
  • the tumor cells can be easily collected by collecting blood from the animal to be treated.
  • Blood collection can be performed by venous blood collection using, for example, a vacuum blood collection tube or a syringe.
  • target tumor cells can be collected.
  • a target tumor cell can be separated using a blood cell separation device used for blood donation.
  • HTLV-1-infected ATL cells present HTLV-1-derived tax as an antigen, it is also possible to isolate ATL cells by FACS or the like using an anti-tax antibody.
  • the vector of the present invention grows only in cells in which the promoter of a gene specifically expressed in hematopoietic tumor cells such as survivin gene can exert a transcriptional activity of a certain level or more and exhibits a cell killing effect
  • the tumor cell may be infected with the vector of the present invention in a state where other blood cells are mixed without isolating the target tumor cell.
  • the collected hematopoietic tumor cells may be amplified by culturing before infection with the vector of the present invention.
  • the basal medium used for culturing hematopoietic tumor cells include, for example, BME medium, BGJb medium, CMRL 1066 medium, Glasgow MEM medium, Improved MEM Zinc Option medium, IMDM medium, 199 medium, Eagle MEM medium, ⁇ MEM medium, DMEM
  • the medium is not particularly limited as long as it is a medium that can be used for culturing animal cells, such as a medium, Ham's F12 medium, RPMI 1640 medium, Fischer's medium, and mixed medium thereof.
  • the medium may be a serum-containing medium or a serum-free medium, but a serum-free medium is preferred from the viewpoint of ensuring the safety of cell transplantation by eliminating different components.
  • a serum-free medium means a medium that does not contain unconditioned or unpurified serum, and a medium that contains purified blood-derived components or animal tissue-derived components (for example, growth factors) is serum-free. It shall correspond to the culture medium.
  • serum-free medium examples include serum-free medium supplemented with an appropriate amount (for example, 1 to 20%) of commercially available KNOCKOUT TM SR (KSR), serum-free medium supplemented with insulin and transferrin (for example, N2B27 [50% DMEM / F12, 50% Neurobasal medium (Gibco), 25 ⁇ g / mL insulin, 100 ⁇ g / mL apotransferrin, 6 ng / mL progesterone, 16 ⁇ g / mL putrescine, 5.2 ng / mL sodium selenite, 1 ⁇ B27 (Gibco), 0.1% BSA, 2 mM glutamine], CHO-S-SFM II (GIBCO BRL), Hybridoma-SFM (GIBCO BRL), eRDF Dry Powdered Media (GIBCOBRL), UltraCULTURE TM (BioWhittaker), UltraDOMA TM ( BioWhittaker), UltraCHO TM (BioWhithit
  • mN3 medium Mechanism. Dev. 59, 89 (1996), etc.
  • other components such as amino acids (eg, glutamine, etc.), organic acids (eg, pyruvate, lactic acid, etc.), antioxidants (eg, 2-mercaptoethanol, etc.), cytokines (eg, IL- 2), growth factors (eg, insulin, etc.), vitamins (eg, ascorbic acid, etc.), antibiotics (eg, penicillin, streptomycin), etc. may be contained at appropriate concentrations.
  • amino acids eg, glutamine, etc.
  • organic acids eg, pyruvate, lactic acid, etc.
  • antioxidants eg, 2-mercaptoethanol, etc.
  • cytokines eg, IL- 2
  • growth factors eg, insulin, etc.
  • vitamins eg, ascorbic acid, etc.
  • antibiotics eg, penicillin, streptomycin
  • the incubator used for culturing hematopoietic tumor cells is not particularly limited as long as it is for cell culture.
  • a flask, a flask for tissue culture, a dish, a petri dish, a tissue culture dish, a multi-dish, a microplate, a microplate Well plates, multi-plates, multi-well plates, chamber slides, petri dishes, tubes, trays, culture bags, and roller bottles can be mentioned.
  • the incubator may be non-cell-adhesive or cell-adhesive.
  • a cell-adhesive incubator is one in which the surface of the incubator is coated with a cell-supporting substrate for the purpose of improving the adhesion with cells. Examples of such a cell-supporting substrate include collagen.
  • the culture can be performed, for example, in a CO 2 incubator under an atmosphere having a CO 2 concentration of about 1 to about 10%, preferably about 2 to about 5%, at about 30 to about 40 ° C., preferably about 37 ° C. .
  • the method of infecting hematopoietic tumor cells with the vector of the present invention varies depending on the type of vector.
  • seeding was carried out in the above incubator at about 10 5 to about 10 6 cells / ml.
  • a virus solution is added to the hematopoietic tumor cell so that the multiplicity of infection (MOI) is about 0.1 to about 3000, preferably about 1 to about 1000, more preferably about 3 to about 300.
  • MOI multiplicity of infection
  • Infections can be established by incubating under conditions for about 0.5 to about 24 hours, preferably about 1 to about 12 hours. Incubation for too long is not preferred because cell lysis occurs prior to administration to the treatment subject.
  • a serum medium as a pre-culture medium for hematopoietic tumor cells, it may be preferable to replace the medium with a serum-free medium prior to virus infection in order to prevent a reduction in infection efficiency.
  • a pharmaceutically acceptable carrier such as parenteral preparations, preferably parenterals such as injections, suspensions, drops, etc.
  • parenteral preparations preferably parenterals such as injections, suspensions, drops, etc.
  • parenteral preparations preferably parenterals such as injections, suspensions, drops, etc.
  • parenteral preparations preferably parenterals such as injections, suspensions, drops, etc.
  • parenteral preparations preferably parenterals such as injections, suspensions, drops, etc.
  • the agent of the present invention includes, for example, a buffer (eg, phosphate buffer, sodium acetate buffer), a soothing agent (eg, benzalkonium chloride, procaine hydrochloride, etc.), a stabilizer (eg, human serum albumin, Polyethylene glycol, etc.), preservatives, antioxidants and the like.
  • a buffer eg, phosphate buffer, sodium acetate buffer
  • a soothing agent eg, benzalkonium chloride, procaine hydrochloride, etc.
  • a stabilizer eg, human serum albumin, Polyethylene glycol, etc.
  • the tumor cells infected with the vector of the present invention are formulated as an aqueous suspension
  • the tumor cells are suspended in the aqueous solution at about 1.0 ⁇ 10 6 to about 1.0 ⁇ 10 7 cells / ml. Just make it cloudy.
  • the preparation thus obtained can be safely administered to mammals such as humans because it does not kill normal cells under conditions where hematopoietic tumor cells in the animal to be treated can be infected and killed. can do.
  • the administration method is not particularly limited, it is preferably systemic administration by injection or infusion, and examples thereof include intravenous administration and intraarterial administration.
  • the dose of the therapeutic agent for hematopoietic tumor of the present invention varies depending on the animal species to be administered, the type of target hematopoietic tumor, body weight, symptoms, administration method, etc., but usually in ATL patients (weight 60 kg),
  • ATL patients weight 60 kg
  • about 1.0 ⁇ 10 5 to about 1 ⁇ 10 7 cells are administered as a single ATL cell dose, or about 2 to about 8 times at intervals of about 2 to about 8 weeks. Is convenient.
  • Example 1 Infection and cell killing effect of survivin-dependent growth type adenovirus vector (Surv.CRA) on leukemia cell line
  • As survivin-dependent growth type adenovirus vector Kamizono, J. et al., Cancer Res., 2005 , 65 (12): Surv.CRAwt described in 5284-91 (E1A gene under the control of a mouse survivin promoter fragment consisting of a nucleotide sequence at positions -173 to -19 with the start ATG being +1, CMV immediate early promoter The E1B ⁇ 55K gene under the control of the CMV, and the EGFP gene under the control of the CMV immediate early promoter).
  • a replication-deficient adenovirus containing the EGFP gene lacking the E1 region as a control vector (Ad. ⁇ E1; prepared according to Chen, SH et al., Proc. Natl. Acad. Sci. USA, 92: 2577-2581, 1995)
  • HTLV-1-infected cell lines S1T, Oh13T, Su9T01, K3T, F6T established from peripheral blood mononuclear cells of ATL patients as leukemia cell lines (Arima, N. et al., J. Virol., 65 (12) : 6892-6899, 1991) and MT-2 (Miyoshi, I.
  • Example 2 Examination of correlation between survivin promoter activity and cell killing effect of Surv.CRA
  • E1 deficiency that expresses LacZ gene under the control of survivin promoter
  • a replication-defective adenovirus (Ad.Surv-LacZ; made according to Chen, SH et al., Proc. Natl. Acad. Sci. USA, 1995 (described above)) is infected with each ATL cell line at MOI30 and introduced externally
  • the activity of the survivin promoter inserted into the prepared vector was examined using ⁇ -galactosidase activity as an index. The results are shown in FIG. 4A.
  • Adenoviral Gene Transcription Efficiency indicates the infection rate when adenovirus is infected with MOI30.
  • IC50 value ⁇ -galactosidase activity and cell killing effect
  • Example 3 Surv.CRA replication and expansion of infection to uninfected leukemia cells
  • Surv.CRAwt was infected with K3T cell line with MOI3 and observed over time until 48 hours.
  • Ad. ⁇ E1 was used as a control vector.
  • EGFP positive cells were detected with a flow cytometer. The results are shown in FIG.
  • the present invention it is possible to use a proliferating virus vector that has been limited to local administration to solid tumors as a vector for treating hematopoietic tumors. That is, the present invention is extremely useful in that it provides a novel gene therapy strategy for hematopoietic tumors.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Virology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Chemical & Material Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Biophysics (AREA)
  • Oncology (AREA)
  • Physics & Mathematics (AREA)
  • Molecular Biology (AREA)
  • Plant Pathology (AREA)
  • Epidemiology (AREA)
  • Mycology (AREA)
  • Hematology (AREA)
  • Biochemistry (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne une nouvelle forme pharmaceutique d'un vecteur viral à prolifération régulée, qui permet la thérapie génique pour une tumeur du tissu hématopoïétique à l'aide du vecteur ; et un nouveau moyen thérapeutique pour une tumeur du tissu hématopoïétique à l'aide de la forme pharmaceutique. L'invention concerne spécifiquement un agent thérapeutique pour une tumeur du tissu hématopoïétique, qui comprend un vecteur viral à prolifération régulée portant un acide nucléique qui code pour au moins une protéine nécessaire à la réplication des virus et qui est sous la régulation d'un promoteur d'un gène pouvant être exprimé spécifiquement dans une cellule de la tumeur du tissu hématopoïétique (de préférence un gène de la survivine), et qui est caractérisé en ce qu'il est administré par voie systémique de telle façon que la cellule de la tumeur du tissu hématopoïétique est infectée par le vecteur ; et autres.
PCT/JP2011/057477 2010-03-25 2011-03-25 Thérapie génique pour une tumeur du tissue hématopoïétique par vecteur viral à prolifération régulée portant un promoteur de la survivine WO2011118819A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2010-070942 2010-03-25
JP2010070942A JP2011201813A (ja) 2010-03-25 2010-03-25 サービビンプロモーターを含む増殖制御型ウイルスベクターによる造血器腫瘍の遺伝子治療

Publications (1)

Publication Number Publication Date
WO2011118819A1 true WO2011118819A1 (fr) 2011-09-29

Family

ID=44673359

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2011/057477 WO2011118819A1 (fr) 2010-03-25 2011-03-25 Thérapie génique pour une tumeur du tissue hématopoïétique par vecteur viral à prolifération régulée portant un promoteur de la survivine

Country Status (2)

Country Link
JP (1) JP2011201813A (fr)
WO (1) WO2011118819A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2022516129A (ja) * 2018-12-26 2022-02-24 上海元宋生物技術有限公司 インターフェロンを発現する腫瘍溶解性ウイルス及びその応用

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2014039516A (ja) * 2012-08-23 2014-03-06 Kagoshima Univ シノビオリンプロモーターを含む増殖制御型ウイルスベクター
EP3690055A1 (fr) * 2014-01-14 2020-08-05 Kagoshima University Procédé d'étiquetage d'une cellule responsable de la cancérisation dans des cellules souches

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003025190A1 (fr) * 2001-09-14 2003-03-27 The New Industry Research Organization Agent promoteur de tumeur specifique et utilisation associee
WO2005115476A1 (fr) * 2004-05-25 2005-12-08 Univ Kurume Médicament comprenant comme ingrédient actif un vecteur proliférant contenant un promoteur survivin
JP2007525949A (ja) * 2003-06-03 2007-09-13 セル ジェネシス インコーポレイテッド 自己プロセッシング性ペプチド切断部位を含む細胞特異的複製適格ウィルス・ベクター
WO2008013276A1 (fr) * 2006-07-27 2008-01-31 Japan Science And Technology Agency Agent thérapeutique pour mésothéliome malin

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003025190A1 (fr) * 2001-09-14 2003-03-27 The New Industry Research Organization Agent promoteur de tumeur specifique et utilisation associee
JP2007525949A (ja) * 2003-06-03 2007-09-13 セル ジェネシス インコーポレイテッド 自己プロセッシング性ペプチド切断部位を含む細胞特異的複製適格ウィルス・ベクター
WO2005115476A1 (fr) * 2004-05-25 2005-12-08 Univ Kurume Médicament comprenant comme ingrédient actif un vecteur proliférant contenant un promoteur survivin
WO2008013276A1 (fr) * 2006-07-27 2008-01-31 Japan Science And Technology Agency Agent thérapeutique pour mésothéliome malin

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
CALDAS H ET AL.: "Survivin-driven therapy for leukemia", BLOOD, vol. 104, no. 11, 2004, pages 752A *
KAMIZONO J ET AL.: "Survivin-Responsive Conditionally Replicating Adenovirus Exhibits Cancer-Specific and Efficient Viral Replication", CANCER RESEARCH, vol. 65, no. 12, 15 June 2005 (2005-06-15), pages 5284 - 5291, XP002997036, DOI: doi:10.1158/0008-5472.CAN-04-2657 *
MORI N ET AL.: "Expression of Survivin in HTLV-I-Infected T-Cell Lines and Primary ATL Cells", BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, vol. 282, no. 5, 20 April 2001 (2001-04-20), pages 1110 - 1113 *
RYOICHI HORIE ET AL.: "Ketsueki-kei Akusei Shuyo o Model to shita Zoshoku no Bunshi Kiko ni Motozuku Bunshi Hyoteki Idenshi Chiryo no Kisoteki Kento", KITASATO IGAKU, vol. 33, no. 1, 28 February 2003 (2003-02-28), pages 72 - 76 *
SUZUKI S ET AL.: "Induction of Cell Death in Adult T-Cell Leukemia Cell Lines by Survivin- Responsive Conditionally Replicating Adenoviruses", HAEMATOLOGICA -THE HEMATOLOGY JOURNAL, vol. 95, no. SUPP.2, June 2010 (2010-06-01), pages 169 - 170 *
TAKURO WATANABE ET AL.: "Kumikae Adenovirus Vector o Mochiita Seijin T-saibosei Hakketsubyo no Saibo Tokuiteki, Bunshi Hyoteki Idenshi Chiryo no Kisoteki Kento", PROCEEDINGS OF THE JAPANESE CANCER ASSOCIATION, vol. 60, 26 September 2001 (2001-09-26), pages 615 *
WU YH ET AL.: "Reversal of drug resistance by silencing Survivin gene expression in acute myeloid leukemia cells", ACTA BIOCHIMICA POLONICA, vol. 55, no. 4, 2008, pages 673 - 680 *
XIE M ET AL.: "A novel triple-regulated oncolytic adenovirus carrying PDCD5 gene exerts potent antitumor efficacy on common human leukemic cell lines", APOPTOSIS, vol. 14, no. 9, September 2009 (2009-09-01), pages 1086 - 1094, XP019727348, DOI: doi:10.1007/s10495-009-0373-3 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2022516129A (ja) * 2018-12-26 2022-02-24 上海元宋生物技術有限公司 インターフェロンを発現する腫瘍溶解性ウイルス及びその応用
JP7378840B2 (ja) 2018-12-26 2023-11-14 上海元宋生物技術有限公司 インターフェロンを発現する腫瘍溶解性ウイルス及びその応用

Also Published As

Publication number Publication date
JP2011201813A (ja) 2011-10-13

Similar Documents

Publication Publication Date Title
Olson et al. Examination of mesenchymal stem cell-mediated RNAi transfer to Huntington's disease affected neuronal cells for reduction of huntingtin
Sonabend et al. Mesenchymal stem cells effectively deliver an oncolytic adenovirus to intracranial glioma
Zhu et al. Oncolytic adenovirus armed with IL-24 inhibits the growth of breast cancer in vitro and in vivo
JP2022506515A (ja) 制御性ｒｎａを発現させるためのベクターシステム
US20150291980A1 (en) Cell capable of producing adeno-associated virus vector
JP2022543445A (ja) ウイルスベクターを含む細胞組成物及び処置方法
US11850215B2 (en) Recombinant adenoviruses and stem cells comprising same
Hamada et al. Adenovirus-mediated transfer of human papillomavirus 16 E6/E7 antisense RNA and induction of apoptosis in cervical cancer
WO2011118819A1 (fr) Thérapie génique pour une tumeur du tissue hématopoïétique par vecteur viral à prolifération régulée portant un promoteur de la survivine
Li et al. E1A-engineered human umbilical cord mesenchymal stem cells as carriers and amplifiers for adenovirus suppress hepatocarcinoma in mice
JP6238485B2 (ja) Ａｕｒｏｒａキナーゼプロモーターを含む増殖制御型ウイルスベクター
CN111500632B (zh) 表达st13和trail的溶瘤腺病毒构建及其应用
US7371570B2 (en) Cell-specific adenovirus vector comprising EBV-specific promoter
JP6483019B2 (ja) 新規アデノウイルス及びその増殖促進方法
JP2022552870A (ja) 改変型アデノウイルスヘキソンタンパク質を有するアデノウイルス
JP5186693B2 (ja) 低酸素状態にある細胞で増殖するウイルスまたは遺伝子を発現するウイルスベクター
JP6099121B2 (ja) 遺伝子導入効率を高めたウイルスベクター及びその利用
NL2031110B1 (en) Recombinant adeno-associated virus vector, recombinant adeno-associated virus aav8-pd1 and use thereof
WO2019183042A1 (fr) Extraction et expansion des mitochondries autologues
WO2020166727A1 (fr) Virus oncolytique utilisant l'adénovirus humain de type 35 comme base
JP5580043B2 (ja) 放射線増感増強剤
Zhu et al. Establishment of a lentiviral vector encoding human HGF and the infection of human ADSCs
Larochelle et al. Modulation of coxsackie and adenovirus receptor expression for gene transfer to normal and dystrophic skeletal muscle
CN117004574A (zh) 改造的免疫细胞、靶向KMT5A基因的gRNA和应用
CN117025674A (zh) 一种重组溶瘤病毒及制备方法与应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 11759623

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 11759623

Country of ref document: EP

Kind code of ref document: A1