KR102692742B1 - miRNA biomarker for diagnosis of pseudoexfoliation glaucoma and uses thereof - Google Patents

Abstract

The present invention relates to a miRNA biomarker for diagnosing pseudoscale glaucoma and its use. In the pseudoscale glaucoma group compared to the normal group, the expression level of 12 types of miRNAs of the present invention is up-regulated or downregulated by more than 2-fold, so it can be used to diagnose pseudoscale glaucoma. It can be used effectively.

Description

miRNA biomarker for diagnosis of pseudoexfoliation glaucoma and uses thereof {miRNA biomarker for diagnosis of pseudoexfoliation glaucoma and uses thereof}

The present invention relates to a miRNA biomarker for diagnosing pseudoscale glaucoma and its use.

Glaucoma is one of the top three blindness diseases worldwide and causes irreversible optic nerve damage. False scale syndrome is closely related to glaucoma, and is an age-related disease with lesions in the extracellular matrix. Small, white, characteristic fibrillar material is deposited in various intraocular as well as extraocular tissues and can progress. Pseudoscale syndrome is the most common observable cause of secondary glaucoma, and when glaucoma occurs, it is called pseudoscale glaucoma. Pseudoscaly glaucoma has higher intraocular pressure, is less responsive to drugs than primary open-angle glaucoma, progresses faster, and is more difficult to treat. The etiology and development mechanism of false scale syndrome are still not well known. It can be explained that individual differences in genetic factors have some influence on the phenotypic diversity of glaucoma and pseudoscale syndrome and the differences in the prevalence of pseudoscale syndrome worldwide.

The Lysyl oxidase-like 1 (LOXL1) gene has been studied extensively in various ethnic groups for its association with false scale syndrome, and several single nucleotide poly-morphisms (SNPs) have also been studied in various ethnic groups. However, the frequency of disease-related mutations in the control group of the disease-related LOXL1 allele is too high, and common disease-related mutations in all races are not evident.

Additionally, the frequency of LOXL1 disease-related SNPs does not correspond to the prevalence of pseudoscaly syndrome. Therefore, the LOXL1 gene alone is insufficient to explain the occurrence and pathogenesis of false scale syndrome.

MicroRNA is a small non-expressing RNA molecule of about 21 to 23 nucleotides and is known to regulate gene expression. Most downregulation of mRNA is caused by destruction of the target mRNA, and some downregulation occurs during translation into protein. It is involved in various biological functions such as cell differentiation, proliferation, apoptosis, individual differentiation, and metabolism, and has been studied in the pathological process of various diseases such as tumors and obesity, and can be used as a non-invasive biomarker for the diagnosis of various diseases. . A single microRNA regulates hundreds of different target mRNAs, and a single target is regulated by multiple microRNAs. Approximately 60% of human protein-coding genes are regulated by miRNAs.

The aqueous humor is structurally close to the ciliary body and the trabecular meshwork, and there is a possibility of discovering new valuable biomarkers by monitoring changes in extracellular miRNAs (e.g., hsa-let-7b-3p) in the aqueous humor.

In a previous study, microRNA was also detected in human aqueous humor, and recently, microRNA was detected in the aqueous humor of normal people and patients with primary open-angle glaucoma, and the differences were reported. In addition, there is a recent study that detected microRNA in the aqueous humor of patients with pseudoscaly glaucoma and compared the results with the microRNA of the aqueous humor of patients with open-angle glaucoma and normal people. However, this study was not conducted on a single race, but rather on a mixed group of whites and blacks. According to another study, microRNA was detected in the eye tissues of normal people, such as the ciliary body, cornea, and trabecular meshwork. In contrast to mRNA, microRNA has remarkable stability in biofluids. Aqueous humor contains not only genes but also various bioregulatory factors and biomarkers, and cytokines such as TNF-a, IL-6, and IL-10 and growth factors such as VEGF are also detected in the aqueous humor, and several studies have been conducted.

In addition, aqueous humor is a widely used specimen not only for diseases of the anterior segment of the eye, but also for diseases of the posterior part of the eye, such as the vitreous body, retinal diseases, glaucoma, and optic nerve diseases. Recent studies have shown that miRNAs are differentially regulated during the pathogenesis of glaucoma. For example, miR-93-5p targets PTEN and is said to regulate NMDA-induced autophagy in retinal ganglion cells.

Korean Patent No. 1585794 discloses a technology for the prevention or treatment of eye diseases using miRNA, and it is disclosed in microRNA expressed in the aqueous humor of patients with exfoliation glaucoma or primary open-angle glaucoma. Although this is disclosed in Human Molecular Genetics (2018, Vol. 27, No. 7 p1263-1275), the present invention's miRNA biomarker for diagnosing pseudoscale glaucoma and its use have not yet been disclosed.

The present invention was developed in response to the above-mentioned needs, and the present invention provides a miRNA biomarker for diagnosing pseudoscale glaucoma and its use, and the expression level of the miRNA of the present invention is more than twice that in the pseudoscale glaucoma group compared to the normal group. By confirming whether it was up- or down-regulated, the present invention was completed.

In order to achieve the above object, the present invention provides hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR- 6728-5p, hsa-miR-6777-5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e- Provided is a biomarker composition for diagnosing pseudoscale glaucoma containing one or more microRNAs selected from 3p as an active ingredient.

In addition, the present invention relates to hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR-6728-5p, hsa -one or more selected from -miR-6777-5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e-3p A composition for diagnosing pseudoscale glaucoma comprising an agent capable of detecting microRNA is provided.

Additionally, the present invention provides a kit for diagnosing pseudoscale glaucoma comprising the composition.

In addition, the present invention provides hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR- 6728-5p, hsa-miR-6777-5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e- Measuring the expression level of one or more microRNAs selected from 3p; and

It provides a method of providing information for the diagnosis of pseudoscale glaucoma, including the step of comparing the expression level of the miRNA with the expression level of the corresponding miRNA in a normal control sample.

The present invention relates to a miRNA biomarker for diagnosing pseudoscale glaucoma and its use. In the pseudoscale glaucoma group compared to the normal group, the expression level of 12 types of miRNA of the present invention is up- or down-regulated by more than 2 times, so it can be used to diagnose pseudoscale glaucoma. It can be used effectively.

Figure 1 shows the results of screening miRNA expression levels to select 12 types of miRNAs with large expression level changes in the pseudoscale glaucoma patient group among the miRNAs present in the aqueous humor.

The present invention relates to a biomarker composition for diagnosing pseudoexfoliative glaucoma, comprising as an active ingredient at least one microRNA selected from hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR-6728-5p, hsa-miR-6777-5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e-3p.

The hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR-6728-5p, hsa-miR-6777 -5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e-3p are each of SEQ ID NOs: 1 to 12 It is preferable that it consists of a base sequence, but it is not limited to this.

In the present invention, micro RNA (miRNA) refers to 21 to 23 non-coding RNAs that post-transcriptionally regulate gene expression by promoting degradation of target RNA or inhibiting their translation. Not only miRNAs represented by specific base sequences, but also precursors (pre-miRNAs, pri-miRNAs) of the above-mentioned miRNAs, miRNAs with equivalent biological functions, such as homologs (i.e. homologs or orthologs), variants such as genetic polymorphisms, and derivatives.

'Diagnosis' in the present invention refers to determining the susceptibility of an object, that is, a test subject, to a specific disease or condition, determining whether an object currently has a specific disease or condition, and determining whether an object currently has a specific disease or condition. A concept that includes determining the prognosis of an affected subject or therametrics (e.g., monitoring the condition of the subject to provide information about treatment efficacy).

In addition, the present invention relates to hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR-6728-5p, hsa -one or more selected from -miR-6777-5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e-3p It relates to a composition for diagnosing pseudoscale glaucoma comprising an agent capable of detecting microRNA.

The hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR-6728-5p, hsa-miR-6777 -5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e-3p are each of SEQ ID NOs: 1 to 12 It is preferable that it consists of a base sequence, but it is not limited to this.

The agent is an agent that can recognize the microRNA by binding complementary to it or amplify the microRNA, and specific examples include antisense oligonucleotides, primers, or probes that can specifically detect the miRNA. It is not limited to this.

The preparation may be directly or indirectly labeled to measure the expression level of the microRNA. Specifically, the label may include, but is not limited to, ligands, beads, radionuclides, enzymes, substrates, cofactors, inhibitors, fluorescent substances, chemiluminescent substances, magnetic particles, haptens, and dyes. It is not limited to this. As specific examples, the ligands include biotin, avidin, and streptoavidin, the enzymes include luciferase, peroxidase, and beta galactosidase, and the fluorescent substances include fluorescein, coumarin, rhodamine, Includes, but is not limited to, phycoerythrin and sulforodamic acid chloride (Texas red). Most of the known labels can be used as these detectable labels, and a person skilled in the art will be able to select an appropriate label to suit the purpose of the invention.

The term "marker or diagnosis marker" as used in the present invention refers to a substance that can diagnose an individual with pseudoscale glaucoma by distinguishing it from normal cells or a normal individual, and pseudoscale glaucoma progresses or develops compared to normal cells. It includes organic biomolecules such as polypeptides, proteins, or nucleic acids (e.g., mRNA, etc.), lipids, glycolipids, glycoproteins, or sugars (monosaccharides, disaccharides, oligosaccharides, etc.) that show an increase or decrease in cells or organisms. For the purpose of the present invention, the diagnostic marker for pseudoscale glaucoma of the present invention is a miRNA or a fragment of the corresponding miRNA that specifically shows a difference in expression level in cells of the pseudoscale glaucoma group compared to normal cells or tissue cells.

As the term of the present invention, "primer" is a base sequence with a short free 3' terminal hydroxyl group, it can form a base pair with a complementary template and is used for copying the template strand. A short sequence that serves as a starting point. In the present invention, the primers used for amplifying the mRNA of the gene are used in an appropriate buffer under appropriate conditions (e.g., four different nucleotide triphosphates and a polymerization agent such as DNA, RNA polymerase or reverse transcriptase) and appropriate temperature conditions. It can be a single-stranded oligonucleotide that can act as a starting point for template-directed DNA synthesis, and the appropriate length of the primer may vary depending on the purpose of use. The primer sequence does not need to be completely complementary to the polynucleotide of the miRNA or its complementary polynucleotide, but can be used as long as it is sufficiently complementary to hybridize.

The term "probe" as used in the present invention refers to a nucleic acid fragment such as RNA or DNA that is as short as a few bases or as long as several hundreds of bases and can form a specific binding to a gene or mRNA, and is called an oligonucleotide. It can be manufactured in the form of a probe, single stranded DNA probe, double stranded DNA probe, RNA probe, etc., and can be labeled for easier detection. Primers or probes of the present invention can be chemically synthesized using the phosphoramidite solid support method or other well-known methods. These nucleic acid sequences can be modified using many means known in the art. Non-limiting examples of such modifications include methylation, capping, substitution of a native nucleotide with one or more homologues, and modifications between nucleotides, such as uncharged linkages (e.g., methyl phosphonate, phosphotriester, phosphoronucleotide). amidates, carbamates, etc.) or charged linkages (e.g. phosphorothioate, phosphorodithioate, etc.).

Considering mutations with biologically equivalent activity, the base sequence used in the present invention is interpreted to include sequences showing substantial identity with the sequences listed in the sequence list. The term 'substantial identity' means that when the sequence of the present invention and any other sequence are aligned to the maximum extent possible and the aligned sequence is analyzed using an algorithm commonly used in the art, the minimum It refers to a sequence that exhibits 60% homology, more specifically 70% homology, even more specifically 80% homology, and most specifically 90% homology. Therefore, a base sequence having high homology to the base sequences represented by SEQ ID NOs: 1 to 12, for example, having a homology of 70% or more, specifically 80% or more, and more specifically 90% or more. Base sequences should also be interpreted as being included within the scope of the present invention.

Additionally, the present invention relates to a kit for diagnosing pseudoscale glaucoma containing the composition of the present invention.

The kit of the present invention uses the composition for diagnosing pseudoscale glaucoma as hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, and hsa-miR-6717-5p. , hsa-miR-6728-5p, hsa-miR-6777-5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa Pseudoscale glaucoma can be diagnosed by measuring the expression level of one or more microRNAs selected from -miR-548e-3p. Specifically, the kit may be an RT-PCR kit, but is not limited thereto. As a specific example, the kit may be a kit containing essential elements required to perform RT-PCR. For example, the RT-PCR kit includes the hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR -6728-5p, hsa-miR-6777-5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e In addition to each primer specific for one or more microRNAs selected from -3p, a test tube or other suitable container, reaction buffer (pH and magnesium concentration are varied), deoxynucleotides (dNTPs), dideoxynucleotides (ddNTPs), Taq-polymer. It may contain enzymes such as enzymes such as enzymes and reverse transcriptases, DNase, RNAse inhibitors, DEPC-water, sterilized water, etc., and may contain primer pairs specific for DNA, RNA or miRNA used as quantitative controls. You can.

In addition, the present invention provides hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR- 6728-5p, hsa-miR-6777-5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e- Measuring the expression level of one or more microRNAs selected from 3p; and

It relates to a method of providing information for the diagnosis of pseudoscale glaucoma, comprising comparing the expression level of the miRNA with the expression level of the corresponding miRNA in a normal control sample.

The expression level was determined by next generation sequencing (NGS), polymerase chain reaction (PCR), reverse transcription polymerase chain reaction (RT-PCR), real-time polymerase chain reaction (Real-time PCR), and RNase protection assay. It is preferable to measure using one or more methods selected from (RNase protection assay; RPA), microarray, and northern blotting, but is not limited thereto.

Hereinafter, the present invention will be described in more detail using examples. These examples are only for illustrating the present invention in more detail, and it is obvious to those skilled in the art that the scope of the present invention is not limited thereto.

1. Adoption of patient and aqueous humor samples

The aqueous humor sample used in the present invention was collected from patients who had undergone cataract surgery and had no problems after obtaining prior consent.

Six patients with pseudoextraglaucoma (PEX) were managed stably using only topical medications, and seven age-matched control subjects who agreed to participate in the present invention were adopted.

Before cataract surgery, approximately 80 to 120 μl of aqueous humor was obtained using a 30-gauge needle. The process of acquiring aqueous humor was performed in all subjects without trauma, thus ruling out the possibility of contamination with cell remnants or blood. All collected samples (aqueous humor) were immediately transferred to liquid nitrogen and stored in a rapidly frozen state.

Clinical data were retrieved from electronic medical records and collected in a completely anonymized manner. Clinical data collected included age, gender, eye orientation, baseline IOP, topical eye drops used, and ocular comorbidities (Table 1).

The clinical trial conducted in the present invention was conducted in accordance with the principles of the Declaration of Helsinki, was approved by the institutional review board of Gyeongsang National University Changwon Hospital College of Medicine (GNUCH-2019-06-001-002), and received prior approval from all subjects registered in the present invention. A consent form was received. All methods were performed in accordance with relevant guidelines and regulations.

Patient and control information # group age gender Eye Laterality Baseline IOP,
mmHg Topical Medication One PEX G 69 F Right 31 Dorzolamide/timolol 2 PEX G 65 M Left 17 Dorzolamide/timolol, brimonidine 3 PEX G 67 M Right 17 Dorzolamide/timolol, brimonidine, bimatoprost 4 PEX G 55 M Left 35 Brinzolamide/timolol, brimonidine, Tafluprost 5 PEX G 64 M Left 37 Dorzolamide/timolol, brimonidine, travoprost 6 PEX G 83 M Right 20 Dorzolamide/timolol, brimonidine, latanoprost 7 PEX G 57 M Right 36 Brinzolamide, timolol 8 PEX G 89 M Right 25 brimonidine, bimatoprost 9 PEX G 63 F Right 18 Dorzolamide/timolol 10 PEX G 78 M Left 11 Dorzolamide/timolol 11 PEX G 86 F Right 32 Dorzolamide/timolol, brimonidine, latanoprost 12 NTG 56 F Left 13 Latanoprost 13 NTG 57 M Left 13 Latanoprost 14 NTG 55 M Left 17 Dorzolamide/timolol 15 NTG 77 F Right 14 Dorzolamide/timolol 16 NTG 72 F Right 15 Latanoprost 17 NTG 76 F Right 17 Tafluprost 18 NTG 76 F Right 14 Latanoprost 19 NTG 74 F Left 12 Dorzolamide/timolol 20 Control 75 F Left 14 None 21 Control 57 F Right 14 None 22 Control 76 F Right 15 None 23 Control 54 M Left 18 None 24 Control 53 M Right 16 None 25 Control 71 F Right 17 None 26 Control 61 M Right 19 None

IOP; Intraocular pressure, NTG; Normal tension glaucoma, PEX; Pseudoexfoliation, Phaco+PCL; phacoemulsification and posterior intraocular lens insertion, Trab; trabeculectomy

2. RNA isolation

Total RNA was extracted using Trizol LS reagent (Invitrogen, USA) according to the manufacturer's instructions. The quality of RNA was assessed with an Agilent 2100 bioanalyzer using an RNA 6000 Pico Chip (Agilent Technologies, Netherlands), and quantification of RNA was performed using a NanoDrop 2000 spectrophotometer system (Thermo Fisher Scientific, USA).

3. Library preparation and RNA sequencing

Libraries were prepared from RNA from the control and patient groups using the NEBNext Multiplex Small RNA Library Prep kit (New England BioLabs, Inc., Ipswich, MA, USA) according to the manufacturer's instructions.

180 pg of total RNA from each sample was used to ligate 1 μg of adapters, and then cDNA was synthesized using adapter-specific primers and reverse transcriptase. For library amplification, aqueous humor collected from pseudoscale glaucoma patient group and control group. The expression pattern of microRNA was analyzed using RNA sequencing, a NGS (Next Generation Sequencing) technique.

As a result, as shown in Figure 1 and Table 2, among the miRNAs present in the aqueous humor, 12 types of miRNAs with a large change in expression level in the pseudoscale glaucoma patient group compared to the control group were selected, and the expression levels of the 12 selected types of miRNAs were As shown in Table 2 below, it was finally confirmed that the expression level increased or decreased by more than twofold in the pseudoscale glaucoma patient group compared to the control group.

miRNAs whose expression levels were increased or decreased in patients with pseudoscale glaucoma compared to the control group miRNA Sequence (5'->3') order
number Accession Number Fold change (Log2) p-value manifestation
change hsa-miR-30d-5p UGUAAACAUCCCCGACUGGAAG One MIMAT0000245 82.070 0.019 Up hsa-miR-320a CUCCCCUCCGCCUUCUCUUCCCGGUUCUUCCCGGAGUCGGGAAAAGCUGGGUUGAGAGGGCGAAAAAGGAUG 2 MI0000542 27.603 0.032 Up hsa-miR-3156-5p AAAGAUCUGGAAGUGGGAGACA 3 MIMAT0015030 0.430 0.034 Down hsa-miR-4458 GAGCGCACAGAGGUAGGUGUGGAAGAAAGUGAAACACUAUUUUAGGUUUUAGUUACACUCUGCUGUGGUGUGCUG 4 MI0016804 0.198 0.036 Down hsa-miR-6717-5p AGGCGAUGUGGGGAUGUAGAGA 5 MIMAT0025846 0.172 0.031 Down hsa-miR-6728-5p UUGGGAUGGGUAGGACCAGAGGGG 6 MIMAT0027357 0.394 0.047 Down hsa-miR-6777-5p ACGGGGAGUCAGGCAGUGGUGGA 7 MIMAT0027454 0.210 0.018 Down hsa-miR-6834-5p GUGAGGGACUGGGAAUUUGUGG 8 MIMAT0027568 0.391 0.026 Down hsa-miR-6864-5p UUGAAGGGACAAGUCAGAUAUGCC 9 MIMAT0027628 0.401 0.029 Down hsa-miR-6879-5p CAGGGCAGGGAAGGUGGGAGAG 10 MIMAT0027658 0.200 0.011 Down hsa-miR-877-3p UCCUCUUCUCCCUCCUCCCAG 11 MIMAT0004950 0.263 0.001 Down hsa-miR-548e-3p AAAAACUGAGACUACUUUUGCA 12 MIMAT0005874 0.162 0.020 Down

<110> INDUSTRY-ACADEMIC COOPERATION FOUNDATION GYEONGSANG NATIONAL UNIVERSITY <120> miRNA biomarker for diagnosis of pseudoexfoliation glaucoma and uses it <130> PN21322 <160> 12 <170> KoPatentIn 3.0 <210> 1 <211> 22 <212> RNA <213> Homo sapiens <400> 1 uguaaacauc cccgacugga ag 22 <210> 2 <211> 72 <212> RNA <213> Homo sapiens <400> 2 cuccccuccg ccuucucuuc ccgguucuuc ccggagucgg gaaaagcugg guugagaggg 60 cgaaaaagga ug 72 <210> 3 <211> 22 <212> RNA <213> Homo sapiens <400> 3 aaagaucugg aagugggaga ca 22 <210> 4 <211> 75 <212> RNA <213> Homo sapiens <400> 4 gagcgcacag agguaggugu ggaagaaagu gaaacacuau uuuagguuuu aguuacacuc 60 ugcugggug ugcug 75 <210> 5 <211> 22 <212> RNA <213> Homo sapiens <400> 5 aggcgaugug gggauguaga ga 22 <210> 6 <211> 25 <212> RNA <213> Homo sapiens <400> 6 uugggauggu aggaccagag gggsa 25 <210> 7 <211> 23 <212> RNA <213> Homo sapiens <400> 7 acggggaguc aggcagguggu gga 23 <210> 8 <211> 21 <212> RNA <213> Homo sapiens <400> 8 ggagggacu gggauuugug g 21 <210> 9 <211> 24 <212> RNA <213> Homo sapiens <400> 9 uugaagggac aagucagaua ugcc 24 <210> 10 <211> 22 <212> RNA <213> Homo sapiens <400> 10 cagggcaggg aaggugggag ag 22 <210> 11 <211> 21 <212> RNA <213> Homo sapiens <400> 11 uccucuucuc ccuccuccca g 21 <210> 12 <211> 22 <212> RNA <213> Homo sapiens <400> 12 aaaaacugag acuacuuuug ca 22

Claims (9)

hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR-6728-5p, hsa-miR-6777- Contains microRNA consisting of 5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e-3p as an active ingredient. Biomarker composition for diagnosing pseudoscale glaucoma. The method of claim 1, wherein hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR-6728-5p , hsa-miR-6777-5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e-3p, respectively. A biomarker composition for diagnosing pseudoscale glaucoma, characterized in that it consists of the base sequences of SEQ ID NOs: 1 to 12. hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR-6728-5p, hsa-miR-6777- An agent capable of detecting microRNA consisting of 5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p, and hsa-miR-548e-3p. A composition for diagnosing pseudoscale glaucoma, comprising: The method of claim 3, wherein hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR-6728-5p , hsa-miR-6777-5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e-3p, respectively. A composition for diagnosing pseudoscale glaucoma, characterized in that it consists of the base sequences of SEQ ID NOs: 1 to 12. The composition for diagnosing pseudoscale glaucoma according to claim 3, wherein the agent is an antisense oligonucleotide, primer, or probe that binds complementary to the microRNA. A kit for diagnosing pseudoscale glaucoma, comprising the composition of any one of claims 3 to 5. The kit for diagnosing pseudoscale glaucoma according to claim 6, wherein the kit is for RT-PCR, real-time PCR, isothermal PCR, Northern blotting, RNA protection assay, or microarray chip. In whole blood or serum, hsa-miR-30d-5p, hsa-miR-320a, hsa-miR-3156-5p, hsa-miR-4458, hsa-miR-6717-5p, hsa-miR-6728-5p, hsa -miR-6777-5p, hsa-miR-6834-5p, hsa-miR-6864-5p, hsa-miR-6879-5p, hsa-miR-877-3p and hsa-miR-548e-3p of microRNA Measuring the expression level; and
A method of providing information for the diagnosis of pseudoscale glaucoma, comprising: comparing the expression level of the miRNA with the expression level of the corresponding miRNA in a normal control sample. The method of claim 8, wherein the expression level is determined by next generation sequencing (NGS), polymerase chain reaction (PCR), reverse transcription polymerase chain reaction (RT-PCR), or real-time polymerase chain reaction (Real-time). Information for diagnosing pseudoscale glaucoma, characterized in that it is measured by one or more methods selected from PCR), RNase protection assay (RPA), microarray, and northern blotting. How to provide.