KR20190112870A - Composition for preventing or treating neurodegenerative disease comprising miR-383-5p inhibitor with inhibitory effect of cell death in hippocampal neural stem cells - Google Patents

Abstract

The present invention relates to a composition for preventing or treating degenerative neurological disease, comprising as an active ingredient a miR-383-5p inhibitor having an inhibitory effect on neural stem cell death. The present inventors performed the microarray-based genome-wide miRNA analysis using HCN cell-derived total RNAs under the condition of the presence of insulin to confirm the characteristics of the miRNAs. As a result, it was confirmed that when miR-383-5p is inhibited, the survival of hippocampal-derived neural stem cells can be controlled to improve neural stem cell engraftment. Therefore, miR-383-5p inhibitor of the present invention can be usefully used for the treatment of neurodegenerative diseases.

Description

Composition for preventing or treating neurodegenerative disease comprising miR-383-5p inhibitor with inhibitory effect of cell death in hippocampal neural stem cells}

The present invention relates to a composition for preventing or treating neurodegenerative diseases comprising a miR-383-5p inhibitor having an effect of inhibiting neural stem cell death as an active ingredient, and when inhibiting miR-383-5p, By controlling survival, neural stem cell engraftment can be improved.

Stem cell-based therapy for regenerating damaged tissue is a new hope for patients with multiple neurological diseases due to permanent damage to the central nervous system. Thus, pluripotent stem cells found in the adult brain, such as the subventricular zone of the lateral ventricle and the dental gyrus of the hippocampus, are drawing attention. Adult neural stem cells contribute to the functional maturation of adult neurons and to maintain a variety of brain activities, including normal cognitive function. In addition, adult rat brain-derived hippocampal neural (HCN) stem cells have been reported to maintain the properties of pluripotent adult neural stem cells. In addition, HCN cells can differentiate into neurons, astrocytes or oligodendrocytes, as well as their self-renewal capacity. This indicates that adult neural stem cells or HCN cells are highly applicable to regenerative treatment of degenerative neurological diseases.

MicroRNAs (miRNAs or miRs) are double-stranded unencoded RNA molecules with about 22-25 nucleotides (nts). It inhibits target mRNA degradation or target mRNA translation, thereby down-regulating gene expression at post-transcriptional levels. MiRNAs produce 70-100 nts of long primary transcripts by RNA polymerase II, called pri-miRNAs. Subsequently, mature miRNA duplexes are made by ribonuclease Drosha and Dicer. The miRNA thus produced complementarily binds to the target mRNA and induces translation inhibition and mRNA destabilization, thereby regulating gene expression mainly related to cell proliferation, death, cell metabolism, intracellular signaling and cell migration. In addition, miRNA expression patterns are tissue-specific and may be defined as the physiological characteristics of cells in certain genetic or environmental situations. In addition, aberrant expression of miRNA disrupts normal physiological activity and inevitably leads cells into a pathological state. Thus, research on miRNAs is increasingly focused on identifying the functional properties of miRNAs in various human diseases. Nevertheless, the role of miRNA (s) in inducing apoptosis of HCN stem cells has not yet been identified.

United States Patent Publication US2015-0037299 (2015.02.05)

An object of the present invention to provide a composition for inhibiting neural stem cell death comprising a miR-383-5p inhibitor as an active ingredient.

Another object of the present invention to provide a pharmaceutical composition for the prevention or treatment of degenerative neurological disease, comprising miR-383-5p inhibitor as an active ingredient.

Still another object of the present invention is to provide a method for screening a therapeutic agent for degenerative brain neuropathy by measuring the expression level of miR-383-5p in neural stem cells.

It is another object of the present invention to provide a biomarker composition for diagnosing degenerative neurological disease comprising miR-383-5p as an active ingredient.

Another object of the present invention to provide a biomarker composition for detecting neural stem cell death comprising miR-383-5p as an active ingredient.

The present invention provides a composition for inhibiting neural stem cell death comprising a miR-383-5p inhibitor as an active ingredient.

The present invention also provides a pharmaceutical composition for the prevention or treatment of degenerative neurological disease, comprising miR-383-5p inhibitor as an active ingredient.

In addition, the present invention comprises the steps of contacting the test substance to the neural stem cells; Measuring the expression level of miR-383-5p in neural stem cells in contact with the test substance; And selecting a test substance having a reduced expression level of miR-383-5p as compared to a control sample.

In another aspect, the present invention provides a biomarker composition for diagnosing degenerative neurological disease comprising miR-383-5p as an active ingredient.

The present invention also provides a biomarker composition for detecting neural stem cell death comprising miR-383-5p as an active ingredient.

The present invention relates to a composition for preventing or treating degenerative neurological disease, comprising as an active ingredient a miR-383-5p inhibitor having an inhibitory effect on neural stem cell death. The present inventors performed the microarray-based genome-wide miRNA analysis using HCN cell-derived total RNAs under the condition of the presence of insulin to confirm the characteristics of the miRNAs. As a result, it was confirmed that when miR-383-5p is inhibited, the survival of hippocampal-derived neural stem cells can be controlled to improve neural stem cell engraftment. Therefore, miR-383-5p inhibitor of the present invention can be usefully used for the treatment of neurodegenerative diseases.

1 shows cell culture results for miRNA arrays. (A) Insulin is present for a specified time; (B) Insulin is deficient for a specified time.
2 shows a sample preparation procedure for miRNA arrays. (A) Insulin (+) experimental group was prepared with a 24-hour sample, insulin (-) experimental group was prepared from the cells cultured in a medium without Insulin for 12, 24, 48 hours according to the time, triple for each experimental group Samples were prepared (n = 3). (B) As a result of total RNA sample analysis, agarose gel electrophoresis (1%) results are shown. (C) As a result of total RNA sample analysis, the result of Nanodrop analysis [(+) 24H] using a spectrophotometer is shown.
3 shows a list of total RNA samples selected for miRNA arrays.
4 shows Affimetrix Genechip 4.0 version information used in a miRNA array.
5 shows the results of miRNA array analysis. (A) Box plot results, which represent the range of signal intensities obtained in the 12 sets of samples used in the miRNA array, showing similar signal intensities in all experimental sets. (B) As a result of the Pearson correlation plot, correlation analysis between array chips is performed, and when the value is close to 1, it means positive correlation. (C) Summary of array analysis.
6 shows the results of quantification of miR-383-5p via RT-qPCR under insulin deficient conditions.
7 shows the effects of miR-383-5p treatment on apoptosis of rHCN cells. (A) Quantitative apoptosis assay (miRNA concentration: 50 nM), (B) Morphological characteristics (48 hours treatment).
8 shows the effect of anti-miR-383-5p treatment on cell death by miR-383-5p. (A) Quantitative apoptosis assay (miRNA concentration: 50 nM, anti-miRNA concentration: 100 nM, 24 hours treatment), (B) Morphological characteristics.
Figure 9 shows the effect of miR-383-5p treatment on autophagy specific cell death. (A) Quantitative caspase-3 activity assay (miRNA concentration: 50 nM, Dox concentration: 1 μM, 24 hours treatment), (B) immunoblotting assay. *: 0.05> p.

The present invention provides a composition for inhibiting neural stem cell death comprising a miR-383-5p inhibitor as an active ingredient.

Preferably, the miR-383-5p inhibitor may be any inhibitor capable of inhibiting miRNA, but may be an antagomir, an antisense oligonucleotide, or an RNA inhibitor molecule. It is not limited to this. More preferably, the miR-383-5p inhibitor may be anti-miR-383-5p represented by SEQ ID NO: 1 (5'-ccacaaucaccuucugaucug-3 ').

Preferably, the composition may improve the engraftment rate of neural stem cells.

Preferably, the neural stem cells may be hippocampal-derived neural stem cells, but are not limited thereto.

The antagomir or antisense oligonucleotide is characterized by having a sequence complementary to the nucleotide sequence of miR-383-5p. Inhibitors having sequences complementary to these miRNAs bind to miR-383-5p and serve to prevent these miRNAs from working.

Meanwhile, miR-383-5p is represented by SEQ ID NO: 2 (5'-c agaucag aaggugacugugg-3 '; miRBase Accession # MIMAT0003114), and rat-derived miR-383-5p or SEQ ID NO: 3 (5'- agaucag). human-derived miR-383-5p, denoted as aaggugauuguggcu-3 '; miRBase Accession # MIMAT0000738). The underlined part of the sequence (bold letters) is an important part of the targeting of miRNA as seed, which used Targetscan information.

As described above, miR-383-5p has a common sequence in humans and rats, and since the common sequence is an important part of the targeting of miRNA, the miR-385-5p inhibitor is a human It can be applied also in can be expected to be effective in the treatment of neurodegenerative diseases.

The present invention also provides a pharmaceutical composition for the prevention or treatment of degenerative neurological disease, comprising miR-383-5p inhibitor as an active ingredient.

Preferably, the miR-383-5p inhibitor may be any inhibitor capable of inhibiting miRNA, but may be an antagomir, an antisense oligonucleotide, or an RNA inhibitor molecule. It is not limited to this. More preferably, the miR-383-5p inhibitor may be anti-miR-383-5p represented by SEQ ID NO: 1 (5'-ccacaaucaccuucugaucug-3 ').

Preferably, the degenerative neurological disease is Alzheimer's disease, stroke, Parkinson's disease, Huntington's disease, Pick's disease, Amyotrophic lateral sclerosis (ALS). , Creutzfeldt-Jakob disease (CJD), Progressive supranuclear palsy, Multiple system strophy, Olive nucleus- Cerebral-cerebellar atrophy (OPCA), Shy-Drager Shy-Drager syndrome, Essential tremor, Cortico-basal ganlionic degeneration, Diffuse Lewy body disease and Striatonigral degeneration It may be, but is not limited thereto.

The pharmaceutical composition of the present invention may include chemicals, nucleotides, antisenses, siRNA oligonucleotides, and natural extracts as active ingredients. The pharmaceutical compositions or complex preparations of the present invention may be prepared using pharmaceutically acceptable and physiologically acceptable auxiliaries in addition to the active ingredients, which may include excipients, disintegrants, sweeteners, binders, coatings, swelling agents, lubricants. Solubilizers such as lubricants and flavoring agents can be used. The pharmaceutical composition of the present invention may be preferably formulated into a pharmaceutical composition including one or more pharmaceutically acceptable carriers in addition to the active ingredient for administration. Acceptable pharmaceutical carriers in compositions formulated in liquid solutions are sterile and physiologically compatible, including saline, sterile water, Ringer's solution, buffered saline, albumin injectable solutions, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

Pharmaceutical formulation forms of the pharmaceutical compositions of the present invention may be granules, powders, coated tablets, tablets, capsules, suppositories, syrups, juices, suspensions, emulsions, drops or injectable solutions and sustained release formulations of the active compounds, and the like. Can be. The pharmaceutical compositions of the present invention may be administered in a conventional manner via intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, sternum, transdermal, nasal, inhalation, topical, rectal, oral, intraocular or intradermal routes. Can be administered. An effective amount of the active ingredient of the pharmaceutical composition of the present invention means an amount required to prevent or treat a disease. Thus, the type of disease, the severity of the disease, the type and amount of the active and other ingredients contained in the composition, the type of formulation and the age, weight, general health, sex and diet, sex and diet, time of administration, route of administration and composition of the patient. It can be adjusted according to various factors including the rate of secretion, the duration of treatment, and the drug used concurrently. For example, in adults, when administered once or several times a day, the inhibitor of the present invention is administered once or several times a day, when the compound is 0.1ng / kg to 10g / kg, a polypeptide, In the case of proteins or antibodies, 0.1ng / kg ~ 10g / kg, antisense nucleotides, siRNA, shRNAi, miRNA can be administered at a dose of 0.01ng / kg ~ 10g / kg.

In addition, the present invention comprises the steps of contacting the test substance to the neural stem cells; Measuring the expression level of miR-383-5p in neural stem cells in contact with the test substance; And selecting a test substance having a reduced expression level of miR-383-5p as compared to a control sample.

As used to refer to the screening method of the present invention, the term "test material" refers to an unknown candidate used in screening to examine whether it affects the expression level of a gene or affects the expression or activity of a protein. do. The test substance includes, but is not limited to, chemicals, nucleotides, antisense-RNAs, miRNAs, small interference RNAs (siRNAs), and natural extracts.

In another aspect, the present invention provides a biomarker composition for diagnosing degenerative neurological disease comprising miR-383-5p as an active ingredient.

The present invention also provides a biomarker composition for detecting neural stem cell death comprising miR-383-5p as an active ingredient.

Hereinafter, the present invention will be described in detail according to embodiments which do not limit the present invention. The following examples of the present invention are not intended to limit or limit the scope of the present invention only to embody the present invention. Therefore, what can be easily inferred by the expert in the technical field to which this invention belongs from the detailed description and the Example of this invention is interpreted as belonging to the scope of the present invention.

< Experimental Example >

The following experimental examples are intended to provide experimental examples that are commonly applied to each embodiment according to the present invention.

One. HCN  Cell culture

Brain-derived hippocampal neural (HCN) stem cells from adult rats were isolated as previously reported (STEM CELLS 2008; 26: 2602-2610). All culture plates were poly-L-ornithine (10 μg / ml plastic plate and 50 μg / ml glass slide; Sigma-Aldrich, St. Louis, MO) and mouse laminin (5 μg / ml; BD Pharmingen, San Diego, CA ). Ham's F-12 medium (Invitrogen), 1 mM L-glutamine (Invitrogen), 100 μg / ml streptomycin, 100 U / ml penicillin (Invitrogen), 20 ng / ml b-FGF (PeproTech, Rocky Hill, NJ) and 5 Dulbecco's modified Eagle's medium (Invitrogen, Carlsbad, Calif.), 16 mg / L putrescin dihydrochloride (Sigma-Aldrich), 100 mg / L transferrin with self-made N2 supplement containing mg / L insulin (Sigma-Aldrich) HCN cells were cultured in serum-free medium containing (Sigma-Aldrich), 30 nM sodium selenite (Sigma-Aldrich), and 20 nM progesterone (Sigma-Aldrich). The medium was adjusted to pH 7.2 using sodium bicarbonate (1.27 g / L, Sigma-Aldrich). Insulin free media was prepared. Cells were passaged at a ratio of 1: 4-6. Cells were cultured at 37 ° C. with 5% CO ₂ conditions. Insulin-containing media was labeled I (+), and media without insulin was labeled I (-).

2. miRNAs  And cell infections

miRNA mimics are double helix RNA molecules that have the same sequence and the same function as endogenous miR-383-5p. To inhibit the miR mimics involved, Anti-miRs chemically synthesized double helix nucleic acids and introduced them into cells via transfection. Negative controls (miR-Con and anti-miR-Con) were purchased from Genolution (Seoul, Korea) and Dharmacon (Lafayette, CO). For transient transfection, HCN cells were inoculated and miRNAs were transfected using Lipofectamine 2000 (Invitrogen).

3. Total RNA Extraction

Total RNA was extracted from HCN cells incubated for 12, 24 and 48 hours depending on the presence of insulin (FIGS. 1 and 2). In RNA-free microcentrifuge tubes, about 2 × 10 ⁶ cells were treated with 0.5 ml Trizol reagent (Invitrogen). After centrifugation at 12,000 g for 15 minutes at 4 ° C., the supernatant was transferred to a new tube containing 0.2 ml chloroform. The tube was shaken vigorously and allowed to react for 5 minutes at room temperature. After centrifugation, the supernatant was transferred to a new tube containing the same amount of pure isopropyl alcohol and allowed to react for 30 minutes at room temperature. After centrifugation, the supernatant was discarded and 1.0 ml of 70% ethanol was added to the RNA pellet for washing. Ethanol was removed by centrifugation and the remaining RNA pellet was suspended in DEPC-treated RNase-free water. The total RNA quantity and quality obtained were measured via NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, Wilmington, DE) and agarose gel electrophoresis, respectively (FIG. 2).

4. miRNA Microarray  And data analysis

Total RNA samples for miRNA microarray analysis were sent to DNALink (Seoul, Korea) (FIG. 3). The quality and concentration of total RNA samples were confirmed by Agilent 2100 bioanalyzer (Agilent Technologies, Santa Clara, Calif.) And NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific), respectively. The RIN value of the total RNA, indicating the degree of RNA integrity, was 9.8 to 10.0, fully satisfying the minimum value> 8.0 required for microarray analysis. In addition, even in the initial electropherogram profile, the RNA sample was found to contain sufficient low molecular weight RNA. Subsequently, microarrays were performed using Affymetrix GeneChip miRNA 4.0 Arrays (Santa Clara, Calif.), Using FlashTag Labeling Kit for probe-labeling and Affymetrix Expression Console Software for data analysis (FIG. 4). . The total number of mature miRNA probes per array was 30,324, including 728 rat mature miRNAs. After background removal, normalization and detection evaluation, signals were shown (FIG. 5). The statistical significance was P-value <0.05 and fold-change cutoff was> 1.3.

5. miRNA  For real time Reverse transcription  Quantitative PCR (RT- qPCR ) analysis

Quantitative analysis of selected mature miRNA expression levels was performed via RT-qPCR. RT was performed using mir-X miRNA first-strand synthesis kit (Clontech, Mountain View, CA). Q-PCR to quantify miRNA expression was performed using IQ SyBr Green I mix (Bio-Rad, Hercules, CA). Forward 5 'specific PCR primers and 3' mRQ primers complementary to miRNAs were used. PCR conditions are as follows. After 95 ° C., 10 seconds (initial denaturation), 40 cycles were performed at 95 ° C., 5 seconds, and 60 ° C., 20 seconds. U6 RNA was used as an endogenous control.

6. Hoechst  33342 / propidium  After iodide (PI) staining, cell death measurement

Cell survival / death of HCN cells was observed morphologically under light microscopy, and the degree of cell death was measured by Hoechst 33342 / PI staining (Thermo Fisher Scientific). At the designated time, 5 μg / ml Hoechst 33342 and PI solution were added to the culture medium, and cells were maintained at 37 ° C. for 1 hour under 5% CO ₂ conditions. Five fields were randomly photographed under fluorescence microscopy, and then the percentage of PI-positive and Hoechst 33342-positive cells was calculated to determine cell death rate.

Cell death (%) = [PI-positive cell number (red) / Hoechst33342-positive cell number (blue)] × 100

< Example  1> HCN  Stem cell culture microRNA  Effect of Insulin Deficiency on Expression Patterns

Within 1 day after HCN deficiency, HCN cells develop cell shrinkage, cellular process damage, and cell death with delayed proliferation of cells. The degree of cell death became most severe at 2-3 days, and the cells eventually separated from the culture plate (FIG. 1). In order to determine whether specific characteristics in miRNA expression during cell death due to insulin deficiency, miRNA microarray analysis was performed (FIGS. 2 and 4). To minimize experimental variation, three independent tests were performed per culture condition with and without insulin. Signal intensity deviations for each array were significantly reduced after quantile normalization as shown in the box plot (FIG. 5A). Correlation coefficients between microarray data sets were greater than 0.97, indicating good reproducibility, as shown in the correlation scatter plot (FIG. 5B). Total miRNA levels were filtered based on p-value cutoff 0.05. Testing of 728 rat mature miRNAs revealed a significant difference in expression levels in total of 175 miRNAs in the absence of insulin as compared to the addition of insulin. After insulin deficiency, 30 miRNAs were up-regulated and 145 miRNAs were down-regulated (fold-change> 1.3, FIG. 5C). Taken together, the results indicate that there is a difference in miRNA expression profile during HCN cell death following insulin deficiency.

As a result of the miRNA array, an increase group of 1.5 or more increased from 6 to 21 species over time, and targets decreased from 0.66 to 57 to 109 species over time. Of these, additional experiments were conducted by selecting miR-383-5p.

Detailed information on miR-383-5p used in the present invention is shown in the table below.

* Blue text: Seeds are important for miRNA targeting (using Targetscan information)

* The other part of sequence between human / rat is marked in red.

* The entire sequence part uses miRBase information.

< Example  2> under insulin deficiency conditions, RT- qPCR  through miR Quantification of -383-5p

The increase observed in the array was confirmed again by qPCR. rHCN cells were incubated for 24 or 48 hours depending on the presence of insulin and RT-qPCR was performed specifically for the indicated miRNAs (n ≧ 3). Expression levels of miR-383-5p were enhanced in insulin deficient conditions (FIG. 6). Each figure is expressed as mean ± SD. U6 RNA was used as an endogenous control.

< Example  3> HCN  cell On viability  Yes, up-regulated miR Effect of -383-5p Treatment

On HCN cells, cells were transfected and observed with synthetic miRNA mimics in the presence of insulin to characterize the physiological role of up-regulated miR-383-5p. In contrast to the control with the control miRNA mimic, cells in the presence of up-regulated miRNA mimics significantly lost viability, confirmed by a significant increase in PI-positive cells.

The degree of cell death by miR-383-5p was similar to that of insulin deficiency (FIG. 7). The results indicate that up-regulated miR-383-5p may be involved in the regulation of HCN cell viability in the absence of insulin.

< Example  4> miR Anti-miR-383-5p inhibits -383-5p-induced cell death and protects against cell death induced by insulin deficiency

To verify the specificity of miR-383-5p in HCN cell death, we used anti-miR-383-5p having complementary sequence with miR-383-5p as a miR-383-5p inhibitor, Knock down the physiological activity of miR-383-5p (FIG. 8). Treatment with anti-miR-383-5p and miR-383-5p simultaneously reduced cell death significantly. Anti-miR-Con with nonspecific mixed sequences did not affect cell death induced by miR-383-5p mimic.

< Example  5> Self-feeding ( autophagy Anti-induced specific cell death miR -383-5p

In the killing induction effect of rHCN cells by miR-383-5p treatment, caspase-3 activity (A) and immunoblotting (B) assays were performed to analyze signal transduction pathways. As a result, referring to Figure 9, while miR-383-5p treatment does not affect the caspase-3 activity, it was confirmed that the caspase-3 activity is increased by the control doxorubicin (Dox) treatment. In addition, the treatment of miR-383-5p effectively induces the conversion from LC3-I to LC3-II, which is a representative marker of autophagy, so that cell death by miR-383-5p is not caused by apoptosis. Specific cell death was confirmed.

<110> University of Ulsan Foundation For Industry Cooperation          THE ASAN FOUNDATION           <120> Composition for preventing or treating neurodegenerative disease          comprising miR-383-5p inhibitor with inhibitory effect of cell          death in hippocampal neural stem cells <130> ADP-2018-0081 <160> 3 <170> KopatentIn 2.0 <210> 1 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> anti-miR-383-5p <400> 1 ccacaaucac cuucugaucu g 21 <210> 2 <211> 21 <212> RNA <213> rat <400> 2 cagaucagaa ggugacugug g 21 <210> 3 <211> 22 <212> RNA <213> human <400> 3 agaucagaag gugauugugg cu 22

Claims (11)

Neural stem cell death inhibition composition comprising miR-383-5p inhibitor as an active ingredient. The composition of claim 1, wherein the miR-383-5p inhibitor is an antagomir, an antisense oligonucleotide, or an RNA inhibitor molecule. The composition for inhibiting neural stem cell death according to claim 2, wherein the miR-383-5p inhibitor is anti-miR-383-5p represented by SEQ ID NO: 1. The composition for inhibiting neural stem cell death according to claim 1, wherein the composition improves engraftment rate of neural stem cells. The composition for inhibiting neural stem cell death according to any one of claims 1 to 4, wherein the neural stem cells are hippocampal-derived neural stem cells. A pharmaceutical composition for preventing or treating neurodegenerative neurodegenerative diseases comprising a miR-383-5p inhibitor as an active ingredient. According to claim 6, wherein the miR-383-5p inhibitor is anti-miR-383-5p represented by SEQ ID NO: 1, characterized in that the pharmaceutical composition for preventing or treating neurodegenerative neurological disease. The method of claim 6 or 7, wherein the degenerative neurological disease is dementia, Alzheimer's disease, stroke, Parkinson's disease, Huntington's disease, Pick's disease, Amyotrophic lateral sclerosis Amyotrophic lateral sclerosis (ALS), Creutzfeldt-Jakob disease (CJD), Progressive supranuclear palsy, Multiple system strophy, Olive nucleus-Cerebral-cerebellar atrophy (Olivopontocerebellar OPPO atrophy) ), Shy-Drager syndrome, Essential tremor, Cortico-basal ganlionic degeneration, Diffuse Lewy body disease, and progenitor-blackening degeneration Pharmaceutical composition for the prevention or treatment of degenerative neurological disease, characterized in that any one selected from the group consisting of striatonigral degeneration. Contacting a test substance with neural stem cells;
Measuring the expression level of miR-383-5p in neural stem cells in contact with the test substance; And
A screening method for treating neurodegenerative neurodegenerative disease, comprising screening a test substance having a reduced expression level of miR-383-5p compared to a control sample. Biomarker composition for diagnosing degenerative neurological disease comprising miR-383-5p as an active ingredient. Biomarker composition for detecting neural stem cell death comprising miR-383-5p as an active ingredient.

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