CN110982894A - Reagent for detecting and targeting biomarker C8orf77 and application thereof - Google Patents

Abstract

The invention discloses a reagent for detecting and targeting a biomarker C8orf77 and application thereof, provides application of the reagent for detecting the biomarker C8orf77 in preparation of a product for diagnosing Alzheimer's disease, and simultaneously provides application of a targeted C8orf77 biomarker inhibitor in preparation of a pharmaceutical composition for treating Alzheimer's disease.

Description

Reagent for detecting and targeting biomarker C8orf77 and application thereof

Technical Field

The invention relates to the technical field of biology, and relates to a reagent for detecting and targeting a biomarker C8orf77 and application thereof.

Background

Alzheimer Disease (AD) is a degenerative disease causing severe memory impairment and dyskinesia (Reitz C, Brayne C, Mayeux R. epidemiology of Alzheimer disease) [ J ] Nature reviews neurology.2016; 7(3):137-52), and is mainly characterized by Clinical signs of β -cell deposits such as amyloid, Neuron Fiber Tangles (NFTs), neuron loss in parts such as temporal lobe and hippocampus, particle vacuole degeneration and the like (Mielke MM, Vemuri P, RoccaWA Clinical epidemiology of Alzheimer's disease, Clinical signs of disease, Clinical diseases, Clinical.

Several studies have shown that alterations in epigenetic and environmental components (Erster S, Moll UM. stress-induced P53 fungi a direct mitochodrial death program: its role in physiologenic and pathophysiologenic stress in vivo [ J ] Cell cycle.2004; 3:1492-5.), changes in the Metabolic status of cholesterol in vivo (Feldhaus P, Fraga DB, lymphoma of tissues Gheiim FV, oral analysis of respiratory tract infection with insulin strain 2011; 26:229-36.) the mechanisms of mitochondrial dysfunction and apoptosis and the mechanisms of insulin signaling and energy metabolism (Jayaksk β. A. the main pathogenesis of this disease, clinical trial protein, AD, leading to the pathogenesis of pathological conditions, including the pathogenesis of vascular disease, Metabolic disease, clinical trial protein, et al, see the pathogenesis of tumor Cell death, clinical trial protein, et al, and the pathogenesis of Metabolic diseases of insulin, clinical disease, clinical trial protein, et al.

Although numerous signaling mechanisms play an important role in the pathogenesis of AD, and pathological lesions continue to develop throughout the disease (Maccioni RB, Rojo LE, Fernandez JA, et al. the role of neurommuneration in Alzheimer's disease, [ J ] Annals of the New YorkAcadmayment of sciences.2009; 1153: 240-6.). However, each of the above-presented hypotheses does not fully account for all known histopathological and biological behavioral changes in AD. Due to the complexity of the pathogenesis of AD, how to prevent and treat it becomes an important research goal in the biomedical field. In recent years, the research of genes in nervous system diseases and various tumors has been advanced, and the research of the correlation between genes and diseases has important significance for disclosing the pathogenesis of diseases and effectively preventing and treating diseases.

Disclosure of Invention

In order to remedy the deficiencies of the prior art, it is an object of the present invention to provide a biomarker associated with alzheimer's disease and its use in the diagnosis and treatment of alzheimer's disease.

In order to achieve the purpose, the invention adopts the following technical scheme:

the invention provides the use of reagents for the determination of genes associated with Alzheimer's disease, including the biomarker C8orf77, in the manufacture of a product for the diagnosis of Alzheimer's disease.

Further, the expression level of biomarker C8orf77 is increased in alzheimer's patients compared to the normal control expression level of biomarker C8orf 77.

Further, the expression level of the biomarker C8orf77 was determined at the transcriptional level.

Further, the expression level of the biomarker C8orf77 was determined by detecting hybridization of a probe to the gene transcript of the biomarker C8orf77 or by amplifying the gene transcript of the biomarker C8orf 77.

Further, the hybridization step is performed in a nucleic acid microarray chip or a microfluidic assay plate.

Further, the amplification reaction includes Polymerase Chain Reaction (PCR).

Further, the polymerase chain reaction includes real-time quantitative polymerase chain reaction.

Further, the primer sequence of the real-time quantitative polymerase chain reaction is shown as SEQ ID NO. 1-2.

Further, the expression level is detected in a biological sample selected from the group consisting of blood, serum and plasma.

The invention provides a product for diagnosing Alzheimer's disease, which comprises a reagent for detecting a biomarker C8orf 77.

The invention provides application of C8orf77 as a molecular target in preparing a pharmaceutical composition for treating Alzheimer disease.

Further, the pharmaceutical composition comprises an inhibitor of biomarker C8orf 77.

Further, the inhibitor reduces the expression of C8orf 77.

Further, the inhibitor is siRNA.

Further, the sequence of the siRNA is shown in SEQ ID NO. 5-6.

The present invention provides a pharmaceutical composition for the treatment of alzheimer's disease comprising an inhibitor of the biomarker C8orf 77.

Further, the inhibitor is siRNA.

Further, the sequence of the siRNA is shown in SEQ ID NO. 5-6.

Further, the pharmaceutical composition also comprises a pharmaceutically acceptable carrier.

The invention has the advantages and beneficial effects that:

the invention discovers that the expression of the C8orf77 gene is related to the Alzheimer disease for the first time, and the Alzheimer patient can be effectively distinguished from the healthy control by detecting the expression level of the C8orf77 in a subject sample.

According to the invention, based on the expression up-regulation of C8orf77 in an Alzheimer patient, siRNA aiming at C8orf77 is designed, and the discovery that the proliferation activity and apoptosis rate of AD cells can be changed by changing the expression level of C8orf77 in cells suggests that C8orf77 has a good application prospect as a molecular target.

Drawings

FIG. 1 is a graph showing the detection of the expression of C8orf77 gene in the blood of Alzheimer's disease patients by QPCR;

FIG. 2 is a graph showing the detection of the expression of C8orf77 gene by QPCR;

FIG. 3 is a graph showing the effect of C8orf77 gene expression on the growth of Alzheimer's nerve cells using MTT;

FIG. 4 is a graph showing the effect of C8orf77 gene expression on Alzheimer's nerve cell apoptosis by flow cytometry.

Detailed Description

The term or "marker" or "biomarker" generally refers to a molecule, including a gene, mRNA, protein, carbohydrate structure, or glycolipid, whose expression or secretion in/on a tissue or cell can be detected by known methods (or methods disclosed herein) and is predictive or used to predict (or help predict) the risk of a patient to develop a disease. A biomarker of particular interest herein is C8orf 77.

The C8orf77 gene is located on region 4 of chromosome 8 long arm 2, and the gene ID in GeneBank, the international public nucleic acid database, is 286103. In the present invention, C8orf77 includes wild type, mutant type or fragment thereof. For the purposes of the present invention, "C8 orf 77" refers to DNA or mRNA encoding C8orf77, including fragments or portions of either which facilitate detection of C8orf77 in a sample. A representative C8orf77 gene has the sequence shown in NR _ 026974.1.

"patient sample" refers to a collection of similar cells obtained from an Alzheimer's patient. The source of the tissue or cell sample may be a solid tissue, like from a fresh, frozen and/or preserved organ or tissue sample or biopsy sample or punch sample; blood or any blood component; body fluids such as cerebrospinal fluid, amniotic fluid (amniotic fluid), peritoneal fluid (ascites), or interstitial fluid; cells from the subject at any time. Tissue samples may contain compounds that are not naturally intermixed with tissue in nature, such as preservatives, anticoagulants, buffers, fixatives, nutrients, antibiotics, and the like. Examples of patient samples herein include, but are not limited to, serum or plasma, Peripheral Blood Mononuclear Cells (PBMCs), circulating plasma proteins, ascites fluid, patient primary cell cultures or cell lines, and preserved tissue samples, such as formalin-fixed, paraffin-embedded tissue samples or frozen tissue samples. In a preferred embodiment, the sample is blood.

The "amount" or "level" of a biomarker associated with clinical benefit in an alzheimer's patient is a detectable level in a biological sample. These can be measured by methods known to those skilled in the art.

The terms "level of expression" or "expression level" are generally used interchangeably and generally refer to the amount of a polynucleotide, mRNA or amino acid product or protein in a biological sample. "expression" generally refers to the process by which information encoded by a gene is converted into structures present and operating in a cell. Thus, according to the present invention, "expression" of a gene may refer to transcription into a polynucleotide, translation into a protein, or even post-translational modification of a protein. Fragments of the transcribed polynucleotide, of the translated protein, or of the post-translationally modified protein should also be considered expressed, whether they are derived from transcripts generated or degraded by alternative splicing, or from post-translational processing of the protein (e.g., by proteolysis). "expressed gene" includes genes that are transcribed into a polynucleotide (e.g., mRNA) and then translated into protein, as well as genes that are transcribed into RNA but not translated into protein (e.g., miRNA, IncRNA).

An "elevated" or "higher" amount or level of a biomarker refers to an amount equal to or greater than the level of expression of the biomarker in a healthy control population that is at least 1.5 fold, e.g., at least 1.6 fold, 1.7 fold, 1.8 fold, 1.9 fold, 2.0 fold, 2.1 fold, 2.2 fold, 2.3 fold, 2.4 fold, 2.5 fold, 2.6 fold, 2.7 fold, 2.8 fold, 2.9 fold, 3.0 fold, 3.1 fold, 3.2 fold, 3.3 fold, 3.4 fold, or 3.5 fold or more over the level of expression of the biomarker relative to the level of control expression.

By "reduced" or "lower" amount or level of a biomarker is meant that the amount is less than the median amount of the biomarker in a healthy control population, the biomarker is underexpressed relative to the control expression level by at least 1.5 fold, e.g., at least 1.6 fold, 1.7 fold, 1.8 fold, 1.9 fold, 2.0 fold, 2.1 fold, 2.2 fold, 2.3 fold, 2.4 fold, 2.5 fold, 2.6 fold, 2.7 fold, 2.8 fold, 2.9 fold, 3.0 fold, 3.1 fold, 3.2 fold, 3.3 fold, 3.4 fold, or 3.5 fold or more.

The terms "nucleic acid" and "polynucleotide" are used interchangeably herein to refer to deoxyribonucleotides or ribonucleotides and polymers thereof in either single-or double-stranded form. The term includes nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, have similar binding properties to the reference nucleic acid, and are metabolized in a similar manner to the reference nucleic acid. Examples of such analogs include, but are not limited to, phosphorothioate, phosphoramide, methylphosphonate, chiral methylphosphonate, 2-O-methyl nucleotide, Peptide Nucleic Acid (PNA).

Unless otherwise indicated, a particular nucleic acid sequence also includes conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences, as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is replaced with mixed base and/or deoxyinosine residues. The term nucleic acid is used interchangeably with gene, cDNA, mRNA, oligonucleotide, and polynucleotide.

The present invention may utilize any method known in the art for determining gene expression. These techniques include, but are not limited to: nucleic acid sequencing, nucleic acid hybridization, nucleic acid amplification techniques, it will be understood by those skilled in the art that the means by which gene expression is determined is not an important aspect of the present invention.

A pharmaceutical composition as used herein, comprising an effective amount of said inhibitor of C8orf77, and a pharmaceutically acceptable carrier, said pharmaceutical composition being useful for treating alzheimer's disease.

As a preferred mode of the invention, the inhibitor of C8orf77 is a small interfering RNA against C8orf 77. As used herein, the term "small interfering RNA" refers to a short segment of double-stranded RNA molecule that targets mRNA of homologous complementary sequence to degrade a specific mRNA, which is the RNA interference (RNA interference) process. Small interfering RNA can be prepared as a double-stranded nucleic acid form, which contains a sense and an antisense strand, the two strands only in hybridization conditions to form double-stranded. A double-stranded RNA complex can be prepared from the sense and antisense strands separated from each other. Thus, for example, complementary sense and antisense strands are chemically synthesized, which can then be hybridized by annealing to produce a synthetic double-stranded RNA complex.

The medicament of the present invention may be in a form suitable for administration by injection, in a form suitable for oral ingestion (e.g., capsules, tablets, caplets, elixirs), in the form of an ointment, cream or lotion suitable for topical administration, in a delivery form suitable for use as eye drops, in an aerosol form suitable for administration by inhalation (e.g., by intranasal or oral inhalation), in a form suitable for parenteral administration, i.e., subcutaneous, intramuscular or intravenous injection.

The pharmaceutical compositions of the invention can also be used in combination with other drugs for the treatment of alzheimer's disease, and other therapeutic compounds can be administered simultaneously with the main active ingredient, even in the same composition. Other therapeutic compounds may also be administered alone, in a single drug or in a different dosage form than the primary active ingredient.

The present invention will be described in further detail with reference to the accompanying drawings and examples.

The following examples are intended to illustrate the invention only and are not intended to limit the scope of the invention.

The experimental procedures, in which specific conditions are not specified in the examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers.

Example 1 QPCR sequencing verification of C8orf77 Gene expression

1. Sample collection

Blood samples were collected from 49 alzheimer patients and 35 age-matched healthy controls. The included alzheimer patients met the standards set by the national institute of neuropathy and the association of senile dementia (NINCDS-ADRDA), and were excluded from other neurological diseases and common senile diseases by medical history investigation, clinical examination, imaging examination, routine laboratory examination, and the like.

2. Trizol method for extracting RNA in sample

Mixing blood and Trizol in equal volume, standing at room temperature for 5 minutes, adding chloroform in the volume of the Trizol reagent 1/5, fully mixing, standing at room temperature for 5 minutes, centrifuging at 12000g at 4 ℃ for 15 minutes, taking supernate into a new 1.5mL centrifuge tube, adding pre-cooled isopropanol 0.5-1.0 mL/tube, slowly shaking up, and standing at room temperature for 10 minutes. 12000g, centrifuging at 4 ℃ for 10 minutes, discarding the supernatant, adding 1mL of precooled 75% ethanol into a centrifugal tube, 7500g, centrifuging at 4 ℃ for 5 minutes, discarding the supernatant, drying the precipitate with sterile wind until the precipitate is transparent, and adding 10 mu L of enzyme-free water into the precipitate for dissolving.

3. Reverse transcription

2. mu.g of total RNA was subjected to reverse transcription, and 10. mu.l of 2 XTRT Reaction Mix, 2. mu.l of RT Enzyme Mix, and 20. mu.l of ribozyme-free water were added. Reacting at 25 deg.C for 10min, at 50 deg.C for 30min, and at 85 deg.C for 5min, and storing the obtained product cDNA at-20 deg.C for use.

4. QPCR amplification

(1) Primer design, the sequence is as follows:

SEQ ID NO.1F：5’-TCTGCCTTCATAGAACTTGTC-3’；

SEQ ID NO. 2R: 5'-GGAGAGGAATTGTGTAGTGTAG-3' (C8orf77 gene)

SEQ ID NO.3F：5’-AACTCTGGTAAAGTGGATATTG-3’；

SEQ ID NO. 4R: 5'-GGTGGAATCATATTGGAACA-3' (internal reference GAPDH).

(2) Preparing a PCR reaction system: power

Green Master Mix 10. mu.l, forward and reverse primers 0.5. mu.l each, cDNA template 1.0. mu.l, deionized water 8.0. mu.l was added.

10min at 95 ℃ (15 s at 95 ℃, 30s at 60 ℃) multiplied by 40 cycles, drawing a melting point curve at 55-95 ℃, 2-^ΔΔCTThe method is used for relative quantification.

5. Statistical method

The experiment was repeated 3 times, the data were expressed as mean ± sd, and the statistical analysis was performed using SPSS17.0 statistical software, and the difference between the two was considered statistically significant when P <0.05 using the t-test. ROC curve analysis was performed on the variable C8orf77 to determine the diagnostic potency, sensitivity and specificity of the gene.

6. Results

As shown in fig. 1, the expression of C8orf77 gene was significantly up-regulated in the blood of alzheimer patients by about 4.3-fold compared to healthy controls, with the difference being statistically significant (P < 0.05).

ROC curve analysis shows that C8orf77 can be used as a biomarker for diagnosing Alzheimer, the area under the curve is 0.934, and Alzheimer patients can be effectively distinguished from healthy controls.

Example 2 silencing of the C8orf77 Gene

1. Cell culture

Culturing SH-SY5Y cell line in DMEM high-sugar complete medium at 37 deg.C under 5% CO₂And culturing in an incubator with the relative humidity of 90%.

2. Transfection

1) Design to synthesize siRNA against C8orf77, siRNA-C8orf77 and general negative control siRNA-NC were purchased from Shanghai Jima Genenco chemical technology, Inc. The sequence of siRNA against C8orf77 was 5'-UUCAGAUUAGGCAUUUCCGGC-3' for the sense strand (SEQ ID NO.5) and 5'-CGGAAAUGCCUAAUCUGAACU-3' for the antisense strand (SEQ ID NO. 6).

2) Treatment of cells prior to transfection

One day before transfection, 6-well culture plates were seeded with 4X 10⁵Cells/well, cultured for one day in antibiotic-free medium, were transfected when the degree of cell confluence was approximately 50% to 70%.

3) Transfection

The nerve cells were divided into 3 groups, namely a blank control group (SH-SY5Y), a control group (transfection siRNA-NC) and an experimental group (siRNA-C8orf 77). Transfection of the vector was performed using liposome 2000 by the following specific transfection method:

mu.L of siRNA-NC, siRNA-C8orf77 and 7.5. mu.L of Lipo2000 were diluted with 100. mu.L of opti-MEM, respectively, gently mixed, and the dilution was performedMixing the siRNA-NC and siRNA-C8orf77 with Lipo2000 by gentle blowing, incubating at room temperature for 20min to form a compound, adding the compound dropwise into a six-hole plate with fresh culture medium replaced, mixing the mixture with gentle shaking, and adding 5% CO at 37 deg.C₂After culturing for 6h in the culture phase, the culture medium was replaced with a conventional medium containing serum and antibiotic for further 24 h.

3. QPCR detection of transcription level of C8orf77 Gene

1) Trizol method for extracting total RNA of cells

Discarding the culture medium in a 6-well plate, adding 1mL of 1 XPBS, slowly cleaning twice, adding 1mL of TrizoI reagent into each well, uniformly mixing, repeatedly blowing vortex cells by using a pipette gun, transferring the obtained cell suspension into a 1.5mL centrifuge tube, standing for 5 minutes at room temperature, adding 200 muL/tube of chloroform, violently shaking and shaking uniformly to be milky white, standing for 5 minutes at room temperature, centrifuging for 15 minutes at 12000g at 4 ℃, taking the supernatant into a new 1.5mL centrifuge tube, adding 0.5-1.0 mL/tube of isopropanol precooled in advance, slowly shaking uniformly, and standing for 10 minutes at room temperature. 12000g, centrifuging at 4 ℃ for 10 minutes, discarding the supernatant, adding 1mL of precooled 75% ethanol into a centrifugal tube, 7500g, centrifuging at 4 ℃ for 5 minutes, discarding the supernatant, drying the precipitate with sterile wind until the precipitate is transparent, and adding 10 mu L of enzyme-free water into the precipitate for dissolving.

2) Reverse transcription and QPCR the same as example 1

4. Statistical method

The experiment was repeated 3 times, the data were expressed as mean ± sd, and the differences between the silenced C8orf77 genome and the control group, which were statistically analyzed using SPSS17.0 statistical software, were considered statistically significant when P <0.05 using the t-test.

5. Results

The results are shown in fig. 2, in which the expression level of C8orf77 gene was significantly reduced (P <0.05) in the experimental group transfected with siRNA-C8orf77 compared to the siRNA-NC group and the blank control group, without significant difference between the siRNA-NC group and the blank control group.

Example 3 Effect of C8orf77 Gene on nerve cells

MTT and apoptosis experiments are adopted to detect the influence of the C8orf77 gene on the cell survival rate of SH-SY5Y Alzheimer disease cell models.

1. Grouping cells:

blank control, A β_1-40(16μM Aβ_1-40Treatment 48h) treatment group, using A β_1-40Treated siRNA-NC control group, Experimental group (siRNA-C8orf77)

2. The cell culture transfection procedure was as in example 2.

3. MTT detection of cell viability

The cells were inoculated in a 96-well plate for overnight culture, and after plating for 1d, 15. mu.L of MTT (0.5kg/mLPBS) was added to each well, followed by further incubation for 4h, followed by addition of 200. mu.L of DMSO to each well, shaking for 10min, and standing at room temperature for 10min to dissolve the blue-violet crystals sufficiently. The wells were calibrated to zero and the absorbance of each well at a wavelength of 570nm was measured using a microplate reader. Cell survival was calculated according to the formula: cell survival (%) — value (experimental absorbance-blank absorbance)/(control absorbance-blank absorbance).

4. Flow cytometry detection of apoptosis

Cells were digested with pancreatin, the cell suspension was transferred to a centrifuge tube, centrifuged at 1000rpm for 5min, the supernatant was discarded, the cells were gently resuspended in PBS, and counted. Taking 0.5-1 × 10⁵Centrifuging the resuspended cells at 1000rpm at 4 deg.C for 5min, and discarding the supernatant; adding 1mL of precooled PBS, gently shaking and uniformly mixing to suspend cells, centrifuging at 1000rpm and 4 ℃ for 5min, discarding supernatant, and repeating the step for 2 times; using 200 mu L of Binging buffer solution for carrying out heavy suspension, adding 10 mu L of each of Annexin V-FITC and PI, gently mixing uniformly, and carrying out a light-shielding reaction at 4 ℃ for 30 min; add Binging buffer 300. mu.L and detect using C6 flow cytometer.

5. Statistical method

The experiment was performed in 3 replicates using SPSS17.0 statistical software for statistical analysis, and the difference between the two was considered statistically significant when P <0.05 using the t-test.

6. Results

The results of the MTT experiment (FIG. 3) show that A β is comparable to the control group_1-40After treatment, SH-SY5Y cell activity (P) is remarkably inhibited<0.05), the cell viability damage of the transfection siRNA-C8orf77 group is obviously improved, namely the siRNA-C8orf77 group improves A β_1-40Cell viability of SH-SY5Y after treatment (P)<0.05), suggesting that differential expression of C8orf77 affects the proliferative activity of neural cells.

The results of the apoptosis experiments (FIG. 4) showed that A β was compared with the blank control group_1-40The apoptosis rate of SH-SY5Y cells after treatment is obviously increased, which shows that A β_1-40Can remarkably promote the apoptosis (P) of SH-SY5Y cells<0.05), whereas silencing of siRNA-C8orf77 transfection inhibited A β to some extent compared to NC control group_1-40The induced apoptosis of SH-SY5Y cells shows that the differential expression of C8orf77 influences the apoptosis of nerve cells to a certain extent.

The above description of the embodiments is only intended to illustrate the method of the invention and its core idea. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications will also fall into the protection scope of the claims of the present invention.

Sequence listing

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Claims (10)

1. Use of a reagent for the determination of genes associated with alzheimer's disease for the manufacture of a product for the diagnosis of alzheimer's disease, characterized in that the associated genes comprise the biomarker C8orf 77.

2. The use according to claim 1, wherein the level of expression of biomarker C8orf77 is increased in alzheimer's patient compared to the level of expression of biomarker C8orf77 normal control.

3. Use according to claim 1 or 2, characterized in that the expression level of the biomarker C8orf77 is determined at the transcriptional level.

4. The use according to claim 3, characterized in that the expression level of biomarker C8orf77 is determined by detecting hybridization of a probe to the gene transcript of the biomarker C8orf77 or by amplifying the gene transcript of the biomarker C8orf 77.

5. The use according to claim 4, wherein the amplification reaction comprises a real-time quantitative polymerase chain reaction, preferably, the primer sequence of the real-time quantitative polymerase chain reaction is shown in SEQ ID No. 1-2.

6. Use according to claim 1 or 2, wherein the expression level is detected in a biological sample selected from blood, serum and plasma.

7. A product for diagnosing alzheimer's disease comprising reagents for detecting the biomarker C8orf 77.

Use of C8orf77 as molecular target for the preparation of a pharmaceutical composition for the treatment of alzheimer's disease.

9. The use according to claim 8, wherein the pharmaceutical composition comprises an inhibitor of the biomarker C8orf77, preferably wherein the inhibitor reduces the expression of C8orf77, preferably wherein the inhibitor is an siRNA, preferably wherein the sequence of the siRNA is as shown in SEQ ID No. 5-6.

10. A pharmaceutical composition for treating Alzheimer's disease, which comprises an inhibitor of a biomarker C8orf77, preferably the inhibitor is siRNA, preferably the sequence of the siRNA is shown in SEQ ID No. 5-6, and preferably the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

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