CN112424381A

CN112424381A - SNP marker for diagnosing cerebral aneurysm, comprising single base polymorphism of ARHGAP32 gene

Info

Publication number: CN112424381A
Application number: CN201980005520.2A
Authority: CN
Inventors: 全进平
Original assignee: Hanlin University Industry University Cooperation Group
Current assignee: Hanlin University Technology Holding Co ltd
Priority date: 2019-04-03
Filing date: 2019-12-24
Publication date: 2021-02-26
Also published as: WO2020204312A1; KR102158716B1

Abstract

The present invention relates to an SNP marker composition for diagnosing cerebral aneurysm, which includes SNP of ARHGAP32 gene, a composition for diagnosing cerebral aneurysm, which includes a preparation capable of detecting or amplifying the SNP marker, a sample kit for diagnosing cerebral aneurysm, which includes the composition, and a method for providing information for diagnosing cerebral aneurysm, which uses the composition and the sample kit. According to the SNP marker composition for diagnosing cerebral aneurysm, the sample kit for diagnosing cerebral aneurysm, and the method for providing information for diagnosing cerebral aneurysm of the present invention, a Single Nucleotide Polymorphism (SNP) in the ARHGAP32 gene can be used as a marker for predicting the risk of developing cerebral aneurysm or diagnosing cerebral aneurysm in korean or asian.

Description

SNP marker for diagnosing cerebral aneurysm, comprising single base polymorphism of ARHGAP32 gene

Technical Field

The present invention relates to an SNP marker composition for diagnosing cerebral aneurysm, including an ARHGAP32 gene SNP, a composition for diagnosing cerebral aneurysm, including a preparation for detecting or amplifying the SNP marker, a sample kit for diagnosing cerebral aneurysm, including the composition, and an information providing method for diagnosing cerebral aneurysm, using the composition and the sample kit.

Background

Base sequence diversity (Polymorphism) refers to variations in base sequence with a frequency of more than 1%, of which 90% is Single Nucleotide Polymorphism (SNP). Human gene sequences are 99.9% identical, 1 SNP occurs in about 250-1000 bp, about 2 million SNPs exist in the human genome, and about 21,000 SNPs exist in the gene. Furthermore, mutation of other single bases with a frequency of 1% or less is generally called mutation (mutation).

The nucleotide sequence mutation analysis or SNP can analyze not only a DNA sequence but also a marker, and plays an important role in mining a trigger gene of a disease such as cancer, asthma, diabetes, or judging a correlation with a specific disease. Therefore, analysis of single nucleotide polymorphisms is useful for understanding the human sensitivity to specific human diseases, preventing congenital diseases or treating onset diseases due to differences in nucleotide sequences.

Cerebral aneurysms, in particular intracranial aneurysms, account for approximately 85% of all subarachnoid haemorrhages (subarachnoid haemorrhages), with a mortality rate of up to 50%. Cerebral aneurysms usually cause subarachnoid hemorrhage, which can be confirmed by first taking a brain CT. When there is a suspected subarachnoid hemorrhage but not observed in CT, diagnosis can be made by extracting cerebrospinal fluid mixed with blood by lumbar puncture. In addition, when a treatment plan is prepared for a confirmed case of a cerebral aneurysm rupture, it is necessary to search for a ruptured part and accurately grasp the position, size, shape, number, and the like, and therefore, it is necessary to perform a "cerebrovascular angiography" examination, and in addition, CT angiography, MRI angiography, or the like may be used, but the above-described method has disadvantages in that: not only is it cumbersome, but it is also difficult to predict the pre-onset or prognosis of cerebral aneurysms.

In view of the above, the present inventors have never abandoned the development of a method for efficiently and easily diagnosing the onset of a cerebral aneurysm, predicting the risk of onset, and predicting the prognosis, and as a result, have developed a diagnostic composition that can diagnose and predict a cerebral aneurysm from a specific gene on an SNP allele, and have completed the present invention.

Disclosure of Invention

Problems to be solved

In the 1 st aspect of the present invention, there is provided an SNP marker composition for diagnosing cerebral aneurysm, comprising Single Nucleotide Polymorphism (SNP) rs371331393 of ARHGAP32(Rho GTPase Activating Protein 32) gene.

In the 2 nd aspect of the invention, there is provided an agent comprising a nucleotide sequence that can detect or amplify rs371331393, which rs371331393 is a SNP of the ARHGAP32 gene.

The present invention provides, in accordance with claim 3, a cerebral aneurysm diagnosis kit containing a cerebral aneurysm diagnosis composition.

In the 4 th aspect of the present invention, there is provided an information providing method for diagnosing a cerebral aneurysm, comprising the step of determining an rs371331393 allele, which is an SNP of the ARHGAP32 gene, from a biological sample isolated from an individual.

Means for solving the problems

The following is a detailed description. The various descriptions and embodiments of the present disclosure may also be applied to various other descriptions and embodiments. That is, all combinations of the various components disclosed herein are within the scope of the invention. The scope of the present invention is not to be considered limited by the following detailed description.

Effects of the invention

The SNP marker composition for diagnosing cerebral aneurysm, the sample kit for diagnosing cerebral aneurysm, and the information providing method for diagnosing cerebral aneurysm of the present invention can use Single Nucleotide Polymorphism (SNP) in ARHGAP32 gene as a marker for predicting risk of developing cerebral aneurysm or diagnosis in korean or asian.

Drawings

<1 > development of 13 SNP markers for diagnosing cerebral aneurysm >

Fig. 1 shows a cerebral aneurysm diagnosis process using ARHGAP32 gene SNP.

Figure 2 shows GWAS results in manhattan plots. Red line indicates significant criticality of the whole genome (P-value ═ 5x 10-8).

FIG. 3 shows a 500kb region map (regional plot) of the variant position of rs371331393, which is a SNP of the ARHGAP32 gene. This figure shows the association of rs371331393 and cerebral aneurysm in 250 patients with cerebral aneurysm and 294 control groups (adjusted for age, sex, hypertension, diabetes, hyperlipidemia, smoking and 4 major factors). The X-axis represents the chromosome position (Mb) and the Y-axis represents the value that converts the P-value to-log 10. The purple diamonds show the rs371331393 variant (P ═ 9.3 kholdham 10-27) used to represent the most significant association in the GWAS of the present invention, and the gray circles represent the other SNPs within 500kb of the rs371331393 variant.

FIG. 4A shows a region map (regional plot) of variant positions (+ -500 kb) of SNPrs6741819 of RNF144A gene. This figure shows the association of rs6741819 with cerebral aneurysms in 250 cerebral aneurysm patients and 294 control groups (adjusted for age, sex, hypertension, diabetes, hyperlipidemia, smoking, and 4 major factors). The X-axis represents the chromosome position (Mb) and the Y-axis represents the value that converts the P-value to-log 10. The purple diamonds show other SNPs within 500kb used to represent the rs6741819 variant (P ═ 4.0 kholdi 10-14) in the GWAS of the present invention, and the gray circles show other SNPs within 500kb of the rs6741819 variant.

B of fig. 4 shows the expression level of the rs6741819 variant in accessory kidney tissue (n 175) using single cell ettl. The X-axis represents genotype and the Y-axis represents order-normalized gene expression levels. The reference (Ref)/opposite (Alt) allele of the variant genotype is the same as the macroeffect/microaffect allele.

Fig. 4C shows a region map (regional plot) of variant positions (± 500kb) of the TMOD1 gene SNPrs 1052270. This figure shows the association of rs1052270 with cerebral aneurysms in 250 patients with cerebral aneurysms and 294 control groups (adjusted for age, sex, hypertension, diabetes, hyperlipidemia, smoking and 4 major factors). The X-axis represents the chromosome position (Mb) and the Y-axis represents the value that converts the P-value to-log 10. The purple diamonds show the other SNPs within 500kb used to represent the rs1052270 variant (P ═ 4.0 kholdi 10-14) in the GWAS of the present invention, and the gray circles represent the rs1052270 variant.

Fig. 4D shows the expression level of the rs1052270 variant in testicular tissue (n ═ 225) using single cell ettl. The X-axis represents genotype and the Y-axis represents order-normalized gene expression levels. The reference (Ref)/opposite (Alt) allele of the variant genotype is the same as the macroeffect/microaffect allele.

FIG. 5A shows a map of the region (regional plot) of variant positions (+ -500 kb) of SNPrs6841581 of the EDNRA gene. This figure shows the association of rs6741819 with cerebral aneurysms in 250 cerebral aneurysm patients and 294 control groups (adjusted for age, sex, hypertension, diabetes, hyperlipidemia, smoking, and 4 major factors). The X-axis represents the chromosome position (Mb) and the Y-axis represents the value that converts the P-value to-log 10. The purple diamonds are shown to represent the rs6841581 variant (P ═ 4.0 kholdi 10-14) in the GWAS of the present invention, and the gray circles represent other SNPs within 500kb of the rs6841581 variant.

B and C of fig. 5 show the expression levels of rs6841581 variant in esophagus (mucosa) (B) (358 samples, effect size 0.38) and skin not exposed to sunlight (suprapubic) (C) (335 samples, effect size 0.21, p 1.2 h 10-6) using single cell eQTL. The X-axis represents genotype and the Y-axis represents order-normalized gene expression levels. The reference (Ref)/opposite (Alt) allele of the variant genotype is the same as the macroeffect/microaffect allele.

<2 > diagnosis of cerebral aneurysm Using SNP of LOX (Lysyl oxidase) Gene >

Fig. 6 shows the results of correlation analysis between cerebral aneurysm and 10 SNPs in example 4.

FIG. 7 shows the result of haplotype correlation analysis of 10 SNPs with cerebral aneurysms in example 4.

< 3> diagnosis of cerebral aneurysm by SNP Using SOX17(SRY-box 17) Gene >

Fig. 8 shows the results of correlation analysis between cerebral aneurysm and 4 SNPs in example 4.

Fig. 9 shows the results of the correlation analysis of the cerebral aneurysm rupture and 4 SNPs in example 4.

Detailed Description

Hereinafter, a specific description will be made. The various descriptions and embodiments of the present disclosure may also be applied to various other descriptions and embodiments. That is, all combinations of the various components disclosed herein are within the scope of the invention. The scope of the present invention is not to be considered limited by the following detailed description.

The term "SNP (single nucleotide polymorphism)" used throughout the present specification refers to a single-base polymorphism, which is a genetic polymorphism in a human body due to a high frequency and stability of SNPs occurring due to a general mutation occurring in one of a plurality of DNA bases occurring at a single site of a chromosome and distributed over the entire genome. In the present invention, the "SNP marker" may be used in combination with "SNP".

The term "marker" used throughout the specification of the present invention means a biomarker that can detect a change in a living body, objectively detect a normal or pathological state of a living body, drug reactivity, and the like.

The term "cerebral aneurysm (or" brain aneurysm) ", also known as Intracranial Aneurysm (IA), used throughout the present specification, is a cerebrovascular disease caused by weakening of the wall of a cerebral artery or a cerebral vein, causing local dilatation of a blood vessel or balloon dilatation (balloon). Cerebral aneurysms usually occur in arteries located at the base of the brain called the basilar artery of the brain (Circle of Willis). About 85% of cerebral aneurysms occur in the anterior portion of the basilar artery ring, including the internal carotid artery and its major branches, which supply blood to the anterior and middle portions of the brain. Cerebral aneurysms are classified by size and morphology. The diameter of the small aneurysm (small aneurysm) is less than 15 mm. Larger aneurysms (lager anesrysm) include large (15 to 25mm), giant (25 to 50mm) and super (over 50mm) aneurysms. Saccular aneurysms (saccular aneurysms) are aneurysms with balloon eversion (saccular outpouching), the most common form of cerebral aneurysm. A strawberry aneurysm is a saccular aneurysm with a similar neck or trunk to strawberries. Fusiform aneurysm (fusiform anesystems) refers to an aneurysm without a main stem.

According to an embodiment of the present invention, the SNP marker composition for diagnosing a cerebral aneurysm according to the present invention may diagnose the invention of a cerebral aneurysm, or predict the risk of developing a cerebral aneurysm, or predict the prognosis of a cerebral aneurysm, but is not limited thereto.

The term "diagnosis" as used throughout the specification of the present invention means to determine whether a particular human has developed a cerebral aneurysm.

The term "prediction" used throughout the specification of the present invention means to determine whether or not there is a possibility of occurrence of a cerebral aneurysm in a specific human body, and if there is a possibility of occurrence, to determine whether or not the possibility of occurrence of a cerebral aneurysm is relatively higher than that in an unspecified large number of people.

According to an embodiment of the invention, rs371331393 is an SNP of the ARHGAP32 gene, which may include the 26 th nucleotide of sequence No. 4, and the base allele of rs371331393 may be G or a. According to an embodiment of the present invention, when the rs371331393 base allele is G or the complementary base allele is C, the cerebral aneurysm can be diagnosed as having the onset or the onset risk is predicted to be high. According to an embodiment of the present invention, the whole genome-wide correlation analysis result indicates that rs371331393 is statistically significant (statistical significance 1.00) when diagnosing a cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed using the SNP.

The term "allele" or "allele" as used throughout the specification is intended to refer to multiple forms of a gene located at the same genetic position on homologous chromosomes. A duality gene is used to indicate diversity, e.g., a SNP has more than two alleles, e.g., has two alleles.

According to an embodiment of the present invention, the SNP marker composition for diagnosing cerebral aneurysm may further include: a SNP selected from the group consisting of 1) rs75822236, which is the GBA (gluconerebrosidase) gene; 2) rs112859779, which is a SNP of TCF24(transcription factor 24) gene; 3) rs79134766, which is an SNP of OLFML2A (Olfactomedin Like 2A) gene; 4) rs 13852525217, which is an SNP of the CD163L1(CD163 molecular Like 1) gene; 5) rs74115822, which is a SNP of the CUL4A (cullin 4A) gene; 6) rs75861150, which is a SNP of the LOC102724084 gene; 7) rs116969723, which is a SNP of LRRC3 (leucoine rich repeat conjugation 3) gene; 8) rs6741819, which is a SNP of RNF144A (Ring Finger Protein 144A) gene; 9) rs59626274, which is a SNP of the FLJ45964 gene; 10) rs17688188, which is a SNP of SPCS3(Signal peptide Complex subBunit 3) gene; 11) rs56942085, which is a SNP of LINGO2(Leucine Rich Repeat And Ig Domain containment 2) gene; and 12) rs72835045, which is 1 or more SNPs of the group consisting of SNPs of MINK1(misshapen/Nck-interacting kinase (NIK) -related kinase 1) gene, but is not limited thereto.

According to an embodiment of the invention, rs75822236 is a SNP of the gba (glucocerebrosidase) gene, which may include the 26 th nucleotide of sequence No. 1, and the base allele of rs75822236 may be C or T. According to an embodiment of the present invention, when the base allele of rs75822236 is C or the complementary base allele thereof is G, the cerebral aneurysm can be diagnosed as having the onset or the risk of the onset is predicted to be high. According to an embodiment of the present invention, the whole genome-wide correlation analysis result indicates that rs75822236 is statistically significant (statistical significance 1.00) when diagnosing a cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed using the SNP.

According to an embodiment of the invention, rs112859779 is a SNP of the TCF24 gene, and may include the 26 th nucleotide of sequence number 2, and the base allele of rs112859779 may be C or T. According to an embodiment of the present invention, when the base allele of rs112859779 is C or the complementary base allele thereof is G, it can be diagnosed that the cerebral aneurysm has no onset or is predicted to have a low risk of onset. According to an embodiment of the present invention, the whole genome-wide correlation analysis result indicates that rs112859779 is statistically significant (statistical significance 0.94) when diagnosing a cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed using the SNP.

According to an embodiment of the invention, rs79134766 is an SNP of the OLFML2A gene, and may include the 26 th nucleotide of sequence number 3, and the base allele of rs79134766 may be C or a. According to an embodiment of the present invention, when the base allele of rs79134766 is a or the complementary base allele thereof is T, it can be diagnosed that the cerebral aneurysm has no disease or is predicted to have a low risk of disease. According to an embodiment of the present invention, the whole genome correlation analysis result indicates that rs79134766 is statistically significant (0.96 statistical significance) when diagnosing cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed by using the SNP.

According to an embodiment of the invention, rs 13852525217 is a SNP of said CD163L1 gene, and may comprise the 26 th nucleotide of sequence No. 5, and said base allele of rs138525217 may be C or T. According to an embodiment of the present invention, when the base allele of rs138525217 is C or the complementary base allele thereof is G, the onset of the cerebral aneurysm can be diagnosed or the risk of the onset can be predicted to be high. According to an embodiment of the present invention, the whole genome-wide correlation analysis result indicates that rs138525217 is statistically significant (statistical significance 1.00) when diagnosing cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed using the SNP.

According to an embodiment of the present invention, rs74115822 is a SNP of the CUL4A gene, and may include the 26 th nucleotide of sequence No. 6, and the base allele of rs74115822 may be G or a. According to an embodiment of the present invention, when the base allele of rs74115822 is G or the complementary base allele thereof is C, the disease of the cerebral aneurysm can be diagnosed or the risk of the disease is predicted to be high. According to an embodiment of the present invention, the whole genome correlation analysis result indicates that rs74115822 has a high statistical significance (statistical significance 0.80) when diagnosing a cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed using the SNP.

According to an embodiment of the present invention, rs75861150 is a SNP of said LOC102724084 gene, and may include the 26 th nucleotide of sequence No. 7, and said base allele of rs75861150 may be T or C. According to an embodiment of the present invention, when the base allele of rs75861150 is C or the complementary base allele thereof is G, it can be diagnosed that the cerebral aneurysm does not develop disease or the risk of developing disease is predicted to be low. According to an embodiment of the present invention, the whole genome-wide correlation analysis result indicates that rs75861150 is statistically significant (statistical significance 1.00) when diagnosing a cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed using the SNP.

According to an embodiment of the invention, rs116969723 is a SNP of the LRRC3 gene, and may include the 26 th nucleotide of sequence number 8, and the base allele of rs116969723 may be G or a. According to an embodiment of the present invention, when the base allele of rs116969723 is a or the complementary base allele thereof is T, it can be diagnosed that the cerebral aneurysm has no onset or is predicted to have a low onset risk. According to an embodiment of the present invention, the whole genome correlation analysis result indicates that rs116969723 is statistically significant (statistical significance 0.81) when diagnosing a cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed using the SNP.

According to an embodiment of the invention, rs6741819 is an SNP of the RNF144A gene and may comprise the 26 th nucleotide of sequence number 9, and the base allele of rs6741819 may be C or T. According to an embodiment of the invention, when the base allele of rs6741819 is T or the complementary base allele of rs6741819 is A, the cerebral aneurysm can be diagnosed not to be attacked or the attack risk is predicted to be low. According to an embodiment of the present invention, the whole genome correlation analysis result shows that rs6741819 is statistically significant (statistical significance 0.72) when diagnosing cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed by using the SNP.

According to an embodiment of the invention, rs59626274 is a SNP of the FLJ45964 gene, and may include the 26 th nucleotide of sequence number 10, and the base allele of rs59626274 may be C or T. According to an embodiment of the present invention, when the base allele of rs59626274 is T or the complementary base allele thereof is a, it can be diagnosed that the cerebral aneurysm does not occur or the risk of the occurrence is predicted to be low. According to an embodiment of the present invention, the whole genome-wide correlation analysis result indicates that rs59626274 has a high statistical significance (statistical significance 0.71) when diagnosing a cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed using the SNP.

According to an embodiment of the invention, rs17688188 is an SNP of the SPCS3 gene, and may include the 26 th nucleotide of sequence No. 11, and the base allele of rs17688188 may be G or a. According to an embodiment of the present invention, when the base allele of rs17688188 is a, or the complementary base allele thereof is T, the cerebral aneurysm can be diagnosed as not having the onset or the onset risk is predicted to be low. According to an embodiment of the present invention, the whole genome correlation analysis result indicates that rs17688188 is statistically significant (statistical significance 0.73) when diagnosing cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed using the SNP.

According to an embodiment of the invention, rs56942085 is a SNP of the LINGO2 gene, and may include the 26 th nucleotide of sequence number 12, and the base allele of rs56942085 may be G or a. According to an embodiment of the present invention, when the base allele of rs56942085 is G, or the complementary base allele thereof is C, the cerebral aneurysm may be diagnosed as having the onset or the risk of the onset may be predicted to be high. According to an embodiment of the present invention, the whole genome-wide correlation analysis result indicates that rs56942085 is statistically significant (statistical significance 0.78) when diagnosing cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed using the SNP.

According to an embodiment of the invention, rs72835045 is a SNP of the MINK1 gene, and may include the 26 th nucleotide of sequence number 13, and the base allele of rs72835045 may be G or a. According to an embodiment of the present invention, when the base allele of rs72835045 is a or the complementary base allele thereof is T, it can be diagnosed that the cerebral aneurysm has no onset or is predicted to have a low risk of onset. According to an embodiment of the present invention, the whole genome-wide correlation analysis result indicates that rs72835045 is statistically significant (statistical significance 0.79) when diagnosing a cerebral aneurysm (table 2), and the cerebral aneurysm can be effectively diagnosed using the SNP.

According to an embodiment of the present invention, the SNP marker composition for diagnosing cerebral aneurysm may further include: a polynucleotide consisting of 10 or more consecutive bases or a polynucleotide complementary thereto, which comprises the 205 th base (rs2303656) of sequence number 14 in the lox (lysyl oxidase) gene; a polynucleotide consisting of 10 or more consecutive bases or a polynucleotide complementary thereto, comprising the 488 th base of seq id No. 15 (rs 3900446); a polynucleotide consisting of 10 or more consecutive bases or a polynucleotide complementary thereto, including, but not limited to, base 678 (rs763497) of SEQ ID NO. 15 and combinations thereof.

According to an embodiment of the present invention, when the allele of the 205 th base of SEQ ID NO. 14 is T or the allele of the complementary base thereof is A, it is predicted that: when G is the allele of the 205 th base of SEQ ID NO. 14 or C is the allele of the complementary base thereof, the risk of developing a cerebral aneurysm is low.

According to an embodiment of the present invention, when the 488 th base allele of SEQ ID NO. 15 is G or the complementary base allele thereof is C, it is predicted that: a risk of developing a cerebral aneurysm higher when the 488 th base allele of SEQ ID NO. 15 is A or the complementary base allele thereof is T; or the onset of a cerebral aneurysm can be diagnosed.

According to an embodiment of the present invention, when the allele of the 678 base of SEQ ID NO. 15 is G or the allele of the complementary base thereof is C, it is predicted that: when the allele of the 678 base of SEQ ID NO. 15 is A or the allele of the complementary base thereof is T, the risk of developing a cerebral aneurysm is high; or the onset of a cerebral aneurysm can be diagnosed.

According to an embodiment of the present invention, when the 488 th base allele of SEQ ID NO. 15 is G or the complementary base allele thereof is C, and the 678 th base allele of SEQ ID NO. 15 is G or the complementary base allele thereof is C, it is predicted that: when the 488 th base allele of SEQ ID NO. 15 is A or the complementary base allele thereof is T and the 678 th base allele of SEQ ID NO. 15 is A or the complementary base allele thereof is T, the risk of developing a cerebral aneurysm is high; or the onset of a cerebral aneurysm can be diagnosed.

According to an embodiment of the present invention, the SNP marker composition for diagnosing cerebral aneurysm may further include: a polynucleotide consisting of 10 or more continuous bases or a polynucleotide complementary thereto, which comprises, but is not limited to, base 326 (rs1072737) of sequence No. 38 in the SOX17(SRY-box 17) gene.

According to an embodiment of the invention, it is possible to predict: when the 326 th base allele of SEQ ID NO. 38 is C or the complementary base allele thereof is G, the risk of developing a cerebral aneurysm is higher than when the 326 th base allele of SEQ ID NO. 38 is A or the complementary base allele thereof is T.

Genetic Information of the SNP included in the SNP marker composition for diagnosing cerebral aneurysm according to an embodiment of the present invention may be obtained from a well-known database, for example, GenBank of National Center for Biotechnology Information: NCBI (National Center for Biotechnology Information), etc., but is not limited thereto.

According to an embodiment of the present invention, the SNP marker composition for diagnosing cerebral aneurysm may be used for cerebral aneurysm diagnosis in korean and asian, but is not limited thereto.

According to an embodiment of the present invention, the SNP included in the SNP marker composition for diagnosing cerebral aneurysm is developed through genome-wide association analysis, and can be used to diagnose cerebral aneurysm easily and efficiently by confirming the allele of the SNP.

The term "Genome-wide association study (GWAS)" as used throughout the specification of the present invention refers to a method of detecting the genotype of about 50 to 200 ten thousand single-base polymorphisms and statistically analyzing the association with a disease, which is the first attempted research method of Ozaki (2002) group of Japan physicochemical research institute, and particularly, GWAS research considers that a variation is "related" to a disease when more genetic variation is found in a population with a disease by comparing DNA or SNP of a patient and a healthy person in a manner of analyzing the variation. At present, due to the completion of HapMap project and the rapid development of microarray technology, the whole genome scan related to gene diversity becomes a common work, and the related analysis of hundreds of whole genomes is successfully implemented, leading to the discovery of a plurality of known or newly added gene variants related to cardiovascular diseases, diabetes and other common diseases. In addition, the technology provides a powerful means for identifying gene diversity related to diagnosis of various diseases.

In the 2 nd aspect of the present invention, rs371331393 is an SNP of the ARHGAP32 gene, and a composition for diagnosing cerebral aneurysm, comprising an agent which can be detected or amplified, is provided. The same applies to the cerebral aneurysm diagnostic composition of claim 2 as the same applies to the same repeated matters as in claim 1.

According to an embodiment of the present invention, the composition for diagnosing a cerebral aneurysm may further include: an agent that can detect or amplify a SNP selected from the group consisting of 1) rs75822236, which is a SNP of the GBA gene; 2) rs112859779, which is a SNP of the TCF24 gene; 3) rs79134766, which is an SNP of the OLFML2A gene; 4) rs 13852525217, which is a SNP of the CD163L1 gene; 5) rs74115822, which is a SNP of the CUL4A gene; 6) rs75861150, which is a SNP of the LOC102724084 gene; 7) rs116969723, which is a SNP of LRRC3 gene; 8) rs6741819, which is a SNP of RNF144A gene; 9) rs59626274, which is a SNP of the FLJ45964 gene; 10) rs17688188, which is a SNP of the SPCS3 gene; 11) rs56942085, which is a SNP of LINGO2 gene; and 12) rs72835045, which is 1 or more SNPs of the group consisting of SNPs of MINK1 gene, but is not limited thereto.

The term "an agent that can detect a SNP" used throughout the specification of the present invention refers to an agent that specifically binds to a SNP marker or SNP included in the SNP marker composition to facilitate identification, or that can detect the SNP and amplify, and "an agent that can amplify a SNP marker" refers to an agent that repeatedly replicates a SNP marker or SNP included in the SNP marker composition and increases the number thereof, for example, a primer that specifically amplifies a polynucleotide including the SNP or a probe that can specifically bind, but is not limited thereto.

The primers used for the SNP amplification may be appropriate conditions in an appropriate buffer (for example, 4 different nucleoside triphosphates, a polymerizing agent such as DNA, RNA polymerase or reverse transcriptase) and single-stranded oligonucleotides that function as a column indicating the origin of DNA synthesis at an appropriate temperature, and the appropriate length of the primers may vary depending on the purpose of use, but is usually 15 to 30 nucleotides. Short primer molecules generally require lower temperatures in order to form a mixture of columnar and stable molecules. The primer sequence need not form complete complementarity with the polynucleotide including the SNP, but its complementarity needs to be such that it can be mixed with the polynucleotide including the SNP.

The term "primer" as used throughout the specification of the present invention refers to a short sequence that can function as a columnar chain replication origin by forming a base pair (base pair) with a complementary template (template) using a base sequence having a short free 3'hydroxyl group (free 3' hydroxyl group). Primers can be used as samples for polymerization reactions (i.e., DNA polymerases or reverse transcriptases) in appropriate buffer solvents and temperatures, and DNA synthesis can be turned on in the presence of 4 different nucleoside triphosphates. At this time, PCR conditions, sensitivity and length of the antisense primer may be modified according to the well-known technique in the art.

The probe used for the SNP detection may be a mixed probe or an oligonucleotide that can bind to a nucleic acid complementary strand depending on a specific sequence. The mixing conditions should show significant differences in the mixing intensity between alleles to allow for tight control so that they mix into only one of the alleles. Most preferably, the central portion of the probe of the invention is aligned with the diversity portion of the diversity sequence. Thus, optimal mixed differences between the allelic forms that differ from each other may result. The probe can be applied to a test kit such as a microarray for detecting alleles and diagnosing cerebral aneurysm, a prediction method and the like. Important probes may be labeled for detection, for example, with a radioisotope, a fluorescent compound, a bioluminescent compound, a chemiluminescent compound, a metal chelate or an enzyme. Appropriate labeling of the probe is well known in the art and can be performed by conventional methods.

According to an embodiment of the present invention, the primer or probe may be chemically synthesized by using a phosphoramidite solid support method or other known methods. The nucleic acid sequence may be modified by a variety of techniques well known in the art. Non-limiting examples of such variations are: substitutions are methylation, adsorption, one or more homologs of a natural nucleotide, and modifications between nucleotides, e.g., modifications to uncharged linkers (e.g., methylphosphonates, phosphotriesters, phosphoramidites, carbamates, etc.), or charged linkers (e.g., phosphorothioates, phosphorodithioates, etc.).

According to an embodiment of the invention, rs371331393 is an SNP of the ARHGAP32 gene, and may include the 26 th nucleotide of sequence No. 4, and the allele of the base of rs371331393 may be G or a.

According to an embodiment of the present invention, the composition for diagnosing a cerebral aneurysm may further include: the following formulations of components can be detected or amplified: a polynucleotide consisting of 10 or more consecutive bases including the 205 th base (rs2303656) of the sequence number 14 in the lox (lysyl oxidase) gene or a polynucleotide complementary thereto, a polynucleotide consisting of 10 or more consecutive bases including the 488 th base (rs3900446) of the sequence number 15 or a polynucleotide complementary thereto, a polynucleotide consisting of 10 or more consecutive bases including the 678 th base (rs763497) of the sequence number 15 or a polynucleotide complementary thereto, and a combination thereof, but is not limited thereto.

According to an embodiment of the present invention, the composition for diagnosing cerebral aneurysm further comprises an agent capable of detecting or amplifying: a polynucleotide consisting of 10 or more consecutive bases including 326 th base (rs1072737) of sequence No. 38 in the SOX17(SRY-box 17) gene or a polynucleotide complementary thereto, but not limited thereto.

The present invention provides, in accordance with the 3 rd aspect thereof, a cerebral aneurysm diagnosis cartridge comprising the cerebral aneurysm diagnosis composition. The same applies to the sample cartridge for diagnosing cerebral aneurysm of claim 2 as the same contents as those repeated in

claims

1 and 2.

According to an embodiment of the present invention, the test cartridge may be a PCR test cartridge, an RT-PCR test cartridge, or a DNA chip test cartridge, but is not limited thereto.

According to an embodiment of the invention, the cartridge may comprise, in addition to the SNPs, polynucleotides, cdnas etc. of the invention, other component compositions, solutions or devices according to the analytical method.

According to an embodiment of the present invention, the PCT sample cell for diagnosing cerebral aneurysm may be a sample cell including components necessary for performing PCR. In addition to the polynucleotides, primers or probes specific for the SNPs, the PCR cartridges may further comprise: test tubes or other appropriate containers, reaction buffers (various pH's and magnesium concentrations), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNAse inhibitors, DEPC-water (DEPC-water), and sterilized water.

According to an embodiment of the present invention, the DNA chip cartridge for diagnosing cerebral aneurysm may be a cartridge including essential components for operating a DNA chip, the DNA chip cartridge including: a substrate to which the polynucleotide, primer or probe specific for the SNP is attached, the substrate may include a nucleic acid corresponding to a quantitative comparison region or a fragment thereof.

According to an embodiment of the present invention, the RT-PCR cartridge for diagnosing cerebral aneurysm may be a cartridge including components necessary for performing RT-PCR. In addition to the various primers specific for the SNPs, the RT-PCR kit may further comprise: test tubes or other appropriate containers, reaction buffers (various pH and magnesium concentrations), deoxynucleotides (dNTPs), dideoxynucleotides (ddNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNAse inhibitors, DEPC-water (DEPC-water), and sterilized water.

In the 4 th aspect of the present invention, there is provided an information providing method for diagnosing a cerebral aneurysm, comprising the step of determining the rs371331393 allele of the SNP of the ARHGAP32 gene from a biological sample isolated from an individual. The same applies to the information providing method for diagnosing a cerebral aneurysm of aspect 4 as the same as the contents repeated in aspects 1 to 3.

According to an embodiment of the present invention, the cerebral aneurysm diagnosis according to the present invention may diagnose onset of cerebral aneurysm, or predict risk of onset of cerebral aneurysm, or diagnose prognosis of cerebral aneurysm, but is not limited thereto.

The term "individual" as used throughout the specification of the present invention means all organisms in which a cerebral aneurysm has become diseased or may become diseased, and specific examples may include: the hiccup includes mammals, cultured fishes, etc., including, but not limited to, white rats, monkeys, cows, pigs, mini pigs, livestock, humans, etc.

The term "sample" used throughout the specification of the present invention means a substance derived from an individual in which a cerebral aneurysm has developed or may develop, and specifically, may include, but is not limited to, tissue, cells, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid, urine, and the like. The gene sample may be obtained from a sample thereof, and the type of the gene sample is not limited as long as the base information of the SNP is confirmed from the gene sample, which may include nucleic acids, for example, cDNA synthesized from DNA, mRNA, or mRNA.

According to an embodiment of the present invention, the individual may be a human, and particularly, may be a korean or asian person, but is not limited thereto.

According to an embodiment of the present invention, the method further comprises: a step of analyzing non-genetic (non-genetic) information for risk prediction or diagnosis of the individual, wherein the non-genetic information may be one or more selected from the group consisting of sex, age, hypertension, diabetes, hyperlipidemia, smoking, family history of aneurysm, biochemical scale and clinical scale.

According to an embodiment of the present invention, in the information providing method, the step of determining the base of the SNP may be a step of confirming the genotype of the SNP. The step of confirming the SNP genotype may be carried out by sequencing analysis, for example, sequencing analysis using an automatic base sequence analyzer, pyrosequencing (pyrosequencing), mixing by microarray, PCR-RELP (restriction fragment length polymorphism) method, PCR-SSCP (single strand hybridization) method, PCR-SSO (specific sequence oligonucleotide) method, ASO (allele specific oligonucleotide) hybridization method combining PCR-SSO method and dot hybridization method, TaqMan-PCR method, MALDI-TOF/MS method, RCA (rolling circle amplification) method, RCA (high resolution hybridization) method, primer extension hybridization method, blotting method, dot hybridization method, and the like. Specifically, for example, the step of determining the base of the SNP in the information providing method according to the present invention may directly confirm the allele of the SNP based on the base sequence including the SNP site sorted (sequencing) by nucleic acid isolated from a sample taken from a subject. The base sequence can be sequenced by methods known in the art. For example, the sequencing step of the base sequence may be carried out by: a step of casting a nucleic acid isolated from a sample taken from an individual into a columnar shape, and performing PCR using primers capable of amplifying gene positions including SNP sites; and confirming the sequence of the PCR reaction product by adopting a restriction enzyme cutting method.

According to an embodiment of the invention, rs371331393 is an SNP of the ARHGAP32 gene, and may include the 26 th nucleotide of sequence number 4, further including: and (c) when the allele of rs371331393 is G, diagnosing that the cerebral aneurysm has developed or predicting high risk of developing the cerebral aneurysm.

According to an embodiment of the present invention, the information providing method for diagnosing a cerebral aneurysm may further include: a step of determining alleles of 1 or more SNPs selected from the group consisting of the following SNPs based on a biological sample isolated from an individual: 1) rs75822236, which is a SNP of the GBA gene; 2) rs112859779, which is a SNP of the TCF24 gene; 3) rs79134766, which is an SNP of the OLFML2A gene; 4) rs 13852525217, which is a SNP of the CD163L1 gene; 5) rs74115822, which is a SNP of the CUL4A gene; 6) rs75861150, which is a SNP of the LOC102724084 gene; 7) rs116969723, which is a SNP of LRRC3 gene; 8) rs6741819, which is a SNP of RNF144A gene; 9) rs59626274, which is a SNP of the FLJ45964 gene; 10) rs17688188, which is a SNP of the SPCS3 gene; 11) rs56942085, which is a SNP of LINGO2 gene; and 12) rs72835045, which is a SNP of the MINK1 gene.

According to an embodiment of the present invention, the information providing method for diagnosing a cerebral aneurysm may further include: 1) rs75822236 is a SNP of the GBA gene, when its allele is C; 2) rs138525217 is a SNP of the CD163L1 gene, when its allele is C; 3) rs74115822 is an SNP of CUL4A gene, when its allele is G; 4) rs56942085 is SNP of LINGO2 gene, and when its allele is G, or when two or more kinds of them are combined, it is diagnosed that cerebral aneurysm has onset or high onset risk.

According to an embodiment of the present invention, the information providing method for diagnosing a cerebral aneurysm may further include: 1) rs112859779 is the SNP of TCF24 gene whose allele is T is 2; 2) rs79134766 is an SNP of the OLFML2A gene, when its allele is a; 3) rs75861150 is a SNP of LOC102724084 gene, when its allele is C; 4) rs116969723 is a SNP of LRRC3 gene, when its allele is a; 5) rs6741819 is the SNP for RNF144A gene, when its allele is T; 6) rs59626274 is a SNP of the FLJ45964 gene, when its allele is T; 7) rs17688188 is a SNP of the SPCS3 gene, when its allele is A; 8) rs72835045 is SNP of MINK1 gene, and when its allele is A, or when two or more kinds of them are combined, the step of diagnosing that the cerebral aneurysm has not developed disease or predicting that the risk of developing disease is low.

According to an embodiment of the present invention, the information providing method for diagnosing a cerebral aneurysm further includes: confirming alleles of the 205 th base (rs2303656) of SEQ ID NO. 14, the 488 th base (rs3900446) of SEQ ID NO. 15, the 678 th base (rs763497) of SEQ ID NO. 15, and combinations thereof in the LOX gene from a biological sample isolated from the individual.

According to an embodiment of the present invention, the information providing method for diagnosing a cerebral aneurysm further includes: confirming the 326 th base (rs1072737) allele of sequence number 38 in the SOX17 gene from a biological sample isolated from the individual.

According to an embodiment of the present invention, in the information providing method for diagnosing cerebral aneurysm, the result of the SNP diversity may be statistically processed by a statistical analysis method generally used in the art, and for example, may be analyzed by variables such as continuous variables (continuous variables), classification variables (categorical variables), Odds Ratios (OR), and 95% confidence intervals (confidence, CI) obtained by using t-check (Student's t-test), x2 test (Chi-square test), regression analysis (linear regression analysis), multiple regression analysis (multiple local regression analysis), and integrated analysis (meta-analysis).

[ embodiments of the invention ]

The present invention will be described in more detail below with reference to examples and experimental examples. However, the examples and experimental examples are only for illustrating the present invention, and the scope of the present invention is not limited by the examples and experimental examples.

<1 > development of 13 SNP markers for diagnosing cerebral aneurysm >

Example 1: collecting the experimental group and the comparative group

In order to carry out the present invention, the cohort (cohort) was derived from the stroke database (2015-2018) of saint heart hospital in chun, in order to collect the database, clinical diagnosis and neuroimaging display patients consistent with hemorrhagic or ischemic stroke were called, and in the GWAS study, patients over 18 years old with saccular (saccular) and dispersive (sporadic) cerebral aneurysms were analyzed. In addition, non-saccular aneurysms such as malformed, anatomical, traumatic and infectious aneurysms and family history aneurysms were excluded.

The control group met the following criteria: 1) patients receiving computed tomography or magnetic resonance angiography for headache outpatient or physical examination; 2) without accompanying with the neurological symptoms such as cerebral artery and vein malformation, intracranial hemorrhage or infarction, etc.; 3) family history of the near relatives showing no cerebral aneurysm or subarachnoid hemorrhage; 4) no history of parkinson's disease or alzheimer's disease; and 5) over 18 years old.

In addition, medical records were studied considering various variables such as sex, age, clinical symptoms such as non-ruptured cerebral aneurysm or subarachnoid hemorrhage, hypertension, diabetes, hyperlipidemia, smoking, family history of arteries, and the like, and angiographic variables of the size, position (anterior or posterior circulation), and number (singular or plural) of aneurysms were examined. The study was approved by the clinical trial committee of the saint heart hospital, Chunchuan (No.2016-3, 2017-9).

Example 2: genotyping and sample quality management (sample) quality control)

Genomic DNA Using HiGeneTM Genomic DNA Prep Kit (BIOFACT, field, Korea)Peripheral blood from the experimental and comparative groups. To analyze the genotype of the samples, based on human genome version 19(build 37), including people in southeast Asia, an AxiomTM Asia Precision Medicine area (PMRA) kit (Thermo Fisher Scientific, MA, USA) containing 75 ten thousand SNPs was used, including 5 ten thousand new markers. Of the 798,148 variants including small insertions/deletions (indels), 508,192 variants passed the following standard quality assessment test (genotype callrate 95%, micro-equivalent gene frequency (MAF) 0.01 and genetic balance law (HWE) p-value 1 kholff 10^-6) GWAS was implemented using the mutations passed.

Example 3: statistical analysis

Technical analysis (Descriptive analysis) is presented using the body number (percentage) of individual and category variables, the average being expressed in standard error. In the present invention, after removing 2 missing values, multivariate logistic analysis (multivariate logistic analysis) with 7 interference factors (sex, age, hypertension, diabetes, hyperlipidemia, smoking and 4 major causes) was performed for 250 cerebral aneurysm patients and 294 control groups, which analysis utilized STATA v11.2(STATA corp., TX, USA). In addition, after adjusting the 7 interference factors, in order to confirm the candidate gene variation of brain aneurysm sensitivity under additional genetic effect model, the program of PLINK 1.9 (https://www.cog- genomics.org/plink/1.9/) Whole genome correlation was performedAnd (6) analyzing.

In order to evaluate the reliability of the potential inheritance in GWAS for the purpose of calculating statistical significance (statistics Power), Genetic Power Calculator (GWAS) was used based on the following assumptionshttp://pngu.mgh.harvard.edu/～ purcell/gpc/): probability of cerebral aneurysm incidence of 5%: ratio of ratios (odds-ratio, OR) in additive effect model (additive effect model): d-prime 0.8; experimental-to-comparative ratio of 1: 1.18; and 5 Piper 10^-8(genome-wide significance) class 1 error rate.

Further, in the analysis hereafter, genetic correlations between 151 subarachnoid hemorrhage and 99 non-disruptive cerebral aneurysm patients were evaluated, and, in order to evaluate the degree of Expression of individual variants of genome-wide correlations in GWAS, the single cell Expression quantitative trait locus (eQTL) was analyzed using Genotype-Tissue Expression (GTEx v7) (https:// gtexport. org/home /). Manhattan plots (Manhattan plots) and regional association plots (regional association plots) utilize the "qqman" instruction of R packet v3.4.0, respectively (seehttps://cran.r-project.org/web/packages/qqman) And LocusZoom v1.3(http://locuszoom.org/)。

Test example 1: confirmation of specific characteristics of the experimental and comparative groups

The patients (experimental group) and the control group (non-patients) were divided according to the criteria described in example 1, and their characteristics are shown in table 1 below. Specifically, of 546, 250 were classified Into Aneurysm (IA) patients, and the remaining 296 were classified into non-patients (control group). Between the two groups, there were no statistical differences in occurrence of gender, Hypertension (HTN), Diabetes (DM), hyperlipidemia (hyperlipidemia) and smoking, and the control group was younger (p) than the cerebral aneurysm patients<0.001). Among the cerebral aneurysm patients, those suffering from subarachnoid hemorrhage (SAH) and non-ruptured cerebral aneurysm (UIA) were 151 patients, respectively

(60.4%) and 99 (39.6%). Also, the majority of patients with cerebral aneurysms are aneurysms of the Anterior circulatory system (n 221, 88.4%) occurring in the middle cerebral artery (n 71), Anterior communicating artery (or Anterior cerebral artery) (n 50), internal carotid artery (n 38), and posterior communicating artery (n 53).

[ TABLE 1 ]

Data in the table are expressed as number of patients (%) and mean ± standard error.

P values were evaluated in a multiple logistic regression model.

Principal components were analyzed and predicted in 4 clusters.

*128

Experimental example 2: analysis of results on Whole genome Association analysis (GWAS)

In order to discover SNP markers associated with the onset of cerebral aneurysms, the following experiments were performed.

First, in the experimental group and the comparative group confirmed in experimental example 1, after adjusting the interference factors (age, sex, hypertension, diabetes, hyperlipidemia, smoking) and 4 major components (primary components), genome-wide association analysis (GWAS) was performed to screen out the experimental group and the comparative group, which showed that the p-value was less than 1 and 10^-5A total of 315 SNPs (including indels) of the default association of (2).

Secondly, out of the 315 SNPs, SNPs with a pagewide disequilibrium (linkage disequilibrium) of less than 0.8 were removed, and genome-wide significance threshold (genome-wide significance threshold) of less than 5 × 10 was selected^-8Total 29 SNPs of p-value of (a). The details of the SNPs to be screened are shown in Table 2 below.

[ TABLE 2 ]

*L95,lower 95％confidence interval；OR,odds ratio；U95,upper 95％confidence interval.

M/M, major allele (major allele type)/minor allele (minor allele type)

MAF, dual genotype rate of experimental group (left) versus control group (right) (minor Allele frequency)

C: after modifying age, gender, hypertension, diabetes, hyperlipidemia, smoking and 4 major components, the OR, 95% confidence interval and P-value were predicted by multiple logistic regression analysis

D: statistical significance (Power) of each SNP was determined by using a network-based Genetic Power Calculator (A)http://zzz.bwh.harvard.edu/gpc/cc2.html) The following parameters were used for the prediction. The incidence rate of cerebral aneurysm is 5 percent, MAF in a contrast group, OR and D-prime in an additive effect model are 0.8; 1:1.18 ratio of experimental group to comparative group (296 in comparative group/250 in experimental group ═ 1.18) and 5H 10^-8 Class 1 error rates (genome wide significance).

Pairwise linkage disequilibrium (LD, r)²<0.8) of the 1 SNP shown in (a); rs7964241 and rs56168082 (r)²0.81); rs2440154 and rs2289668 (r)²＝1.0)

In particular, rs371331393 located on ARHGAP32(11q24.3) showed the most significant correlation in the process of cerebral aneurysm formation (OR 43.57; 95% confidence interval 21.84-86.95; p 9.3 KHz 10^-27) (FIG. 3), rs75822236 located on GBA Gene

SNPs also show significant associations in the development of cerebral aneurysms (OR-161.46; 95% CI 53.86-483.60; p-1.1, 10 of Piperk^-19) The variation regressed for the "T" allele in the control group compared to the "C" allele in patients with cerebral aneurysms (MAF 0.01vs 0.33). And rs 13852525217 located in the non-coding region (UTR) of the 3' non-coding region of the CD163L1 geneAmong SNPs, the "T" allele has a low frequency of occurrence and shows a high effect size (MAF experiment/control group 0.31/0.01; OR 75.98; p 6.2 khkal 10)^-23)。

Also, it was confirmed that rs59626274(stop-gain, FLJ45964, 2q37.3) of the additional 4 SNPs [ rs75822236(R535H, GBA, 1q22), rs112859779(G141S, TCF24, 8q13.1), rs79134766(a208T, OLFML2A, 9q33.3), rs3744644(E639D, SCARF1, 17p13.3) ] and the additional human sensitivity SNP have potential direct correlation with the encrypted protein in sequence.

In summary, in the problems related to the invention of cerebral aneurysm, the statistical significance of SNPs located on GBA, TCF24, OLFML2A, ARHGAP32, CD163L1, CUL4A, LOC102724084, and LRRC3 genes was up to 0.8, and the statistical significance of SNPs located on RNF144A, FLJ45964 SPCS3, LINGO2, and MINK1 genes was up to 0.7 to 0.8. The 13 SNPs can be effectively used to predict the risk of cerebral aneurysm onset or diagnose whether cerebral aneurysm onset has occurred.

Experimental example 3: analysis of Single cell eQTL

In order to confirm whether or not the expression has been carried out in the tissues of the SNP markers screened in experimental example 2, the following experiment was performed.

Specifically, 3 SNPs showed significant differences in expression (p) in single-cell eQTL analysis using more than 70 specimens taken from 45 human tissues<1Х10^-5). In particular rs6741819(RNF144A,2p 25.1; p ═ 4.0 KHz 10^-14) Downregulation in accessory kidney tissue was obtained (175 samples, effect size-0.36, p-1.5 KHz 10)^-6) (fig. 4A, B), rs1052270 (tmod1.9q22.33; p2.7 KHz 10^-8) Up-regulation was observed in testis tissue (225 samples, effect size 0.42, p 8.6 KHz 10)^-10) (FIG. 4C, D).

And rs6841581(EDNRA. 4q31.22; p ═ 5.2 KHz 10^-12) And unexposed skin (suprapubic part) (335 samples, effect size 0.21, p 1.2 KHz 10^-6) Were all up-regulated (fig. 5).

<2 > diagnosis of cerebral aneurysm by SNP of LOX (Lysyl oxidase) gene >

Example 1 study object

A single institution was commissioned with 80 patients with cystic (saccular) cerebral aneurysms and 80 control groups matching the age and gender of the patients in the patient group, as confirmed by radiology. Of the cerebral aneurysms, spindle or anatomically non-saccular aneurysms, as well as traumatic or infectious aneurysms, are excluded. The control group consisted of matched patients who received computed tomography or magnetic resonance angiography for headache outpatient or physical examination, except patients with other neurological disorders such as arteriovenous malformations, intracranial hemorrhage or infarction. Medical records were studied considering various variables such as gender, age, clinical symptoms such as non-ruptured cerebral aneurysm or subarachnoid hemorrhage, hypertension, diabetes, hyperlipidemia, smoking, family history of aneurysm, and the like, and examining angiographic variables of size, location (pre-or post-circulation) and number (single or plural) of aneurysm. The study was approved by the clinical trial committee of saint heart hospital, Chunchuan (No. 2016-31).

According to the study results, the number of women in the cerebral aneurysm patient group was 43 (53.8%), and the average age was 57.1 ± 12.9 years. 41 (51.3%) patients developed subarachnoid hemorrhage symptoms. The pre-circulating aneurysm (n-73; 91.3%) was recorded as follows with respect to the aneurysm location: internal carotid artery, n is 15; anterior communicating artery or anterior cerebral artery, n ═ 21; middle cerebral artery, n ═ 25; and a posterior communicating artery, n-12. 75 (93.8%) patients had a single aneurysm, family history of subarachnoid hemorrhage. The incidence of hypertension, diabetes, hyperlipidemia and smoking did not differ significantly between the two groups.

Example 2 Single base diversity screening and detection

In order to examine the correlation between single base diversity on LOX gene and the incidence probability of cerebral aneurysm, linkage disequilibrium (LD;r²<0.8), 10 tag (tag) SNPs located between 5 '-upstream and 3' -downstream of the LOX gene were selected (see SEQ ID NO. 14 to SEQ ID NO. 21).

[ TABLE 3 ]

To analyze the genotype (genotyping) of 10 tagged SNPs, HiGene was used^TMGenomic DNA extraction kit (BIOFACT, Tada, Korea), Genomic DNA was extracted from peripheral blood of 160 subjects. The 10 SNP primers were designed using the primer-3 v.0.4.0 program (http:// bioinfo. ut. ee/primer3-0.4.0/), and the primers were designed as shown in Table 4.

[ TABLE 4 ]

Using the primers and Solg of Table 4 above^TM2X Taq PCR Pre-Mix (Solgent, Tada., Korea) Polymerase Chain Reaction (PCR) was performed. Pre-modification at 95 ℃ for 5 minutes, modification at 95 ℃ for 30 seconds, annealing at 63 ℃ for 30 seconds, stretching at 72 ℃ for 1 minute, final stretching at 72 ℃ for 5 minutes, and the final stretching was repeated 34 times. The amplified fragments were confirmed by 1.5% agarose gel electrophoresis using Solg^TMAfter the PCR refinery cartridge (SolGent, field, Korea) was refined, each sequence was analyzed using ABI PRISM 3730XL Analyzer (Applied Biosystems, Calif., USA).

Example 3 statistical analysis

The non-genetic factor differences between 80 cerebral aneurysms and 80 control groups were assessed by the Kruskal-Wallis test (Kruskal-Wallis). To predict the Odds Ratio (OR), the oppositions between cerebral aneurysms and 10 SNPs adjacent to OR located on the LOX gene were evaluated by the fisher snow exact test.

In subsequent analyses, the genetic effect of the LOX gene on the development of cerebral aneurysms resulting from rupture of the aneurysm was analyzed. And, a likelihood ratio test method is adopted to analyze haplotype specific correlation by a progressive chi-square test statistical method including h-1 degree of freedom (h is the number of all feasible haplotypes in a sliding window according to the number of SNPs), thereby testing the significant haplotype correlation with the cerebral aneurysm. In the assay, 10 SNPs were used, excluding haplotype structures with a mini-effective haplotype frequency (MHF) of less than 0.01 for a feasible combination.

Correlation between monomeric SNPs and haplotypes, significant p-values corrected by pairwise comparison (Bonferroni) were found to be less than 0.005 and less than 2.0x 10 after multiple comparison corrections and default valid p-value detection for 0.01 and 0.001, respectively^-4. STATA software v.11.2(Stata Corp., College Station, Texas, USA) was used for technical and univariate analysis. To evaluate Genotyping Call Rate (GCR), micro-effective dual gene frequency (MAF), genetic equilibrium law (Hardy Weinberg equibrium, HWE) p-value and paired LD (pairwise LD), Haploview v.4.2 (GCR), Haploview V.4.2 (GCR), and the like, were usedhttps://www.broadinstitute.org/haploview/haploview), quality management tests were performed for 10 SNPs. The correlation between genotype and haplotype was analyzed using the PLINK program v.1.07(http:// zzz. bwh. harvard. edu/PLINK /).

Example 4 study of genetic correlation between Single base diversity and cerebral aneurysms

After quality management testing of all 10 SNPs selected in example 2, the call rate for complete Genotyping (GCR), the frequency of the micro-effective dual genes (MAF) exceeded 0.01, the law of genetic balance (HWE) p value exceeded 0.05, and the pairwise Linkage Disequilibrium (LD) exceeded 0.8.

Of the 10 SNPs, 3 SNPs (rs2303656, rs3900446 and rs763497) showed statistically significant association with cerebral aneurysms (p<0.05) (fig. 6), 2 SNPs (rs2303656 and rs3900446) reached significance (p) corrected using pairwise comparison<0.005). The C-allele of rs3900446 showed the most significant strong association with increased risk of cerebral aneurysm (OR 20.15, p 4.8 γ 10)^-5). On the contrary, the a-pair gene of rs2303656 has a cerebral aneurysm protection effect, and the effect is not seen in the patient group (p is 8.2 γ 10)^-4). The G-allele of rs763497 shows a correlation with increased risk of cerebral aneurysms (OR ═ 2.26, p ═ 0.009). However, in the subsequent analysis, no SNP (p) associated with rupture of cerebral aneurysm was observed in 80 patients with cerebral aneurysm>0.05)。

In the likelihood ratio test on haplotype associations, 136 of 247 haplotype structures of 45 sliding windows (SNP groups) showed progressive p-values less than 0.05 and 17 haplotypes showed default associations with cerebral aneurysms (asymptotic p-values)<0.001, fig. 7). Of the 10 SNP haplotype combinations, the CG combination of rs3900446 and rs763497 was analyzed in a single SNP assay (MHF ═ 0.113, asymptote p ═ 1.3 γ 10^-5) Showing significant correlation.

In the single SNP analysis, 6 SNPs (rs10040971, rs17148773, rs3792801, rs10519694, rs2956540 and rs1800449) do not have correlation alone, but the haplotype structure combining the SNPs with rs2303656, rs3900446 or rs763497 has significance in haplotype analysis (6.5 γ 10)^-4<p<7.5Υ10^-4)。

< 3> diagnosis of cerebral aneurysm by SNP Using SOX17(SRY-box 17) Gene >

Example 1 study object

A single institution (chunchuan saint heart hospital) was entrusted with 187 patients with saccular (saccular shape) cerebral aneurysms and 372 patients in the patient group matched in age and sex (ratio of patient group to control group 1: 2). Of the cerebral aneurysms, spindle or anatomically non-saccular aneurysms, as well as traumatic or infectious aneurysms, are excluded. The control group consisted of matched patients who received computed tomography or magnetic resonance angiography for headache outpatient or physical examination, except patients with other neurological disorders such as arteriovenous malformations, intracranial hemorrhage or infarction. Medical records were studied considering various variables such as gender, age, clinical symptoms such as non-ruptured cerebral aneurysm or subarachnoid hemorrhage, hypertension, diabetes, hyperlipidemia, smoking, family history of aneurysm, and the like, and examining angiographic variables of size, location (pre-or post-circulation) and number (single or plural) of aneurysm. The study was approved by the clinical trial committee of saint heart hospital, Chunchuan (No. 2017-9).

According to the study results, the number of women in the cerebral aneurysm patient group and the control group was 108 (57.8%) and 212 (57.9%), respectively, and the mean ages were 58.2 ± 11.5 years and 56.9 ± 14.2 years (P ═ 0.305), respectively. There was no significant difference between the two groups including hypertension, diabetes, hyperlipidemia and other causes of smoking status. 95 (50.8%) patients developed subarachnoid hemorrhage symptoms. The pre-circulating aneurysm (n 167; 89.3%) was recorded as follows with respect to aneurysm location: internal carotid artery, n ═ 34; anterior communicating artery or anterior cerebral artery, n ═ 39; middle cerebral artery, n ═ 56; and the posterior communicating artery, n 38. In this group, no family history of cerebral aneurysms is mentioned.

Example 2 Single base diversity screening and detection

To examine the correlation between single base diversity on SOX17 gene and the possibility of cerebral aneurysm onset, 5 SNPs (rs1072737, rs1504749, rs10958409, rs12541742 and rs9298506) previously reported were screened from the previous GWAS consisting of eastern asian and european cohorts (see sequence No. 38 to 42 of table 5).

[ TABLE 5 ]

To analyze the genotypes (genotyping) of the 5 SNPs, HiGene was used^TMGenomic DNA extractionGenomic DNA was extracted from peripheral blood of 559 subjects in a kit (BIOFACT, Tada, Korea). The 5 SNP primers were designed using the primer-3 v.0.4.0 program (http:// bioinfo. ut. ee/primer3-0.4.0/), and the primers were designed as shown in Table 6.

[ TABLE 6 ]

Using the primers and Solg of Table 6 above^TM2X Taq PCR Pre-Mix (Solgent, Tada., Korea) Polymerase Chain Reaction (PCR) was performed. Pre-modification at 95 ℃ for 5 minutes, modification at 95 ℃ for 30 seconds, annealing at 63 ℃ for 30 seconds, stretching at 72 ℃ for 1 minute, final stretching at 72 ℃ for 5 minutes, and the final stretching was repeated 34 times. The amplified fragments were confirmed by 1.5% agarose gel electrophoresis using Solg^TMAfter the PCR refinery cartridge (SolGent, field, Korea) was refined, each sequence was analyzed using ABI PRISM 3730XL Analyzer (Applied Biosystems, Calif., USA).

Example 3 statistical analysis

In order to evaluate the difference between 187 patients with cerebral aneurysms and 372 patients with age, gender, hypertension, diabetes, hyperlipidemia and smoking, multiple logistic regression analysis (Univariate logistic regression analysis) was performed. To assess the genetic correlation between cerebral aneurysms and SOX17 gene diversity under a synergistic genetic model, a 95% Confidence Interval (CI) and Odds Ratio (OR) were predicted using a general linear model. In subsequent analyses, the genetic effect of SOX17 on the formation of cerebral aneurysms by rupture of the aneurysm was analyzed. Furthermore, the description and univariates were analyzed using Stata II.2(StataCorp LLC, College Station, Texas, USA). In order to evaluate Genotyping Call Rate (GCR), micro-effective dual gene frequency (MAF), genetic equilibrium law (Hardy Weinberg equibrium, HWE) p-value and paired LD (pair LD), a quality management test was performed on 5 SNPs adjacent to or on SOX17 gene using Haploview 4.2(https:// www.broadinstitute.org/Haploview/Haploview). The correlation between genotype and haplotype was analyzed using PLINK program 1.07(http:// zzz. bwh. harvard. edu/PLINK /).

After quality management tests, the complete Genotyping Call Rate (GCR), the micro-effect-pair frequency (MAF) of 4 SNPs out of 5 SNPs screened in example 2 exceeded 10%, the genetic equilibrium law (HWE) p value exceeded 0.05, and the pairwise Linkage Disequilibrium (LD) exceeded 0.8. Due to the law of genetic equilibrium (HWE), the present correlation test removed rs12541742 located in the intron region of the SOX17 gene.

Of the 4 SNPs, only 1 SNP, i.e., only the minor C-mate gene of rs1072737, which is located about 41kb upstream from the 5 '-noncoding region (5' -UTR) of SOX17 gene, showed statistically significant correlation with cerebral aneurysms (OR 0.69, 95% CI 0.49-0.96, P0.03) (fig. 8). This variation showed an opposite effect compared to expression studies including the european ancestor group (OR 0.69 and OR 1.02, respectively). In the subsequent analysis, among 4 SNPs, no SNP associated with cerebral aneurysm rupture was seen (P >0.2, fig. 9).

As described above, it will be understood by those skilled in the art to which the present invention pertains that the present invention can be implemented in other specific embodiments without changing the technical ideas or essential features thereof. In view of the above, it should be understood that all the embodiments described above are only for illustrating the present invention by way of example and the present invention is not limited thereto. All changes and modifications that come within the meaning and range of equivalency of the claims are to be embraced within their scope.

<110> Hanlin University college of labor Cooperation Foundation (Hall University)

<120> SNP marker for diagnosing cerebral aneurysm comprising single base polymorphism of ARHGAP32 gene

<130> 4

<160> 52

<170> KoPatentIn 3.0

<210> 1

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 1

ccttgagtat ctgctccatc actggygacg ccacaggtag gtgtgaatgg a 51

<210> 2

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 2

tgcttcagaa actgaccagt agcacygatg tacaatcttg atcgcatggg c 51

<210> 3

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 3

cctccagctg ctgcagaagg atgccrccgc cgcccctgcc acccctgcca c 51

<210> 4

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 4

ttgtggttca gtcttaaaga ttcttrcccg gatgcagcat attttactcc a 51

<210> 5

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 5

ttcacaagtc ggacatctgt atatcytagg aggagacaag gccatagaag a 51

<210> 6

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 6

gaatccacgt tgaaaaatct gtccarggct cacccagtat caacaatagt g 51

<210> 7

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 7

atatctttct gtgcctttat ttattytgca tatgttaaaa gtggatgagg c 51

<210> 8

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 8

ttcaggaggt ccccgaggac atcccrgcca acaccgtgct cctgaagctc g 51

<210> 9

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 9

tattgaacag atagaacaaa cgggcygcgc tgcccacacc accgcttctc t 51

<210> 10

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 10

gaggcccgag aaccaggcac gccgaygaca gcacacttcc agcctgagag c 51

<210> 11

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 11

acagagaaaa tgtgggaggc aatacrgtga cagtggtgta attaaaatgt t 51

<210> 12

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 12

aagttttgac tattctactg gcctgrattt atctaacttg tgttgtactt t 51

<210> 13

<211> 51

<212> DNA

<213> human (Homo sapiens)

<400> 13

ctcagtctca gggggactgg gaagarggaa ggacaaaagg atggtggccc t 51

<210> 14

<211> 504

<212> DNA

<213> human (Homo sapiens)

<400> 14

tacgcatgat gtcctgtgta gcgaatgtca cagcgcacaa cattgttggt atagtcagat 60

tcaggaacca ggtagctggg gtttacactg acctgggcaa cacaaagagt tcctcagtat 120

ttctttttcc atagggctac aataaggaaa ttgttaatga ggacttagct aaatcaagca 180

gggaagggat tttaacttaa gtaagtggtt aaactctgga gacgttatgg aaagagactg 240

catattttcc cctgaagttc tttaaaataa gacagattag attagattag attgtttata 300

aatagtttga ggatgtataa ttgcttccaa taccatgatt atttaacatt tgaatccaga 360

gaagagggcc tattgatctg caatatcaat atatgatata ttttcaaagg tctttacctt 420

taggatatag tttccaggtt ttacatctgt aatatcaatc cactggcagt ctatgtctgc 480

accataggta tcataacagc cagg 504

<210> 15

<211> 815

<212> DNA

<213> human (Homo sapiens)

<400> 15

gagcatgcaa cctaaatccc tcatatgcac agttcacaat ggggttcacg ctcatatgag 60

aatctaatgc catgattgat ctgacaggaa gcaaagctca ggtggtaatg tgaggaaaga 120

ggatcttggt aaacatgcta ttgtaagttt aatgtctcta ctccccttcc gagaacccca 180

tgtctcctta ctgggaaact tcaagtcaaa tggttttagg atttactgac agaggttact 240

tgctagtatg aatttgagaa attcatggga ggtacctcac caggaaccca aaaaatggaa 300

tttttgacgt ggggagtgtc tgtaaataca gatgaagctt cacttgctgg cccactgctc 360

acctcctgct gtgcagccca gttcctaaca gggatggtac tggtccgtgt cctggggttt 420

gggaaccact ggtttattga aaagacattc tgctgggttg gtatggagcc caagttatat 480

ttgggttaca cattagggac tacttcttgt gattttgagc ctaaattata ataaactcta 540

gatatacaca ccctatgcct atcagaatag taatgaaggg gtaaatggcc cccaacacaa 600

gtggcaatgg tttttggaat aataccaaaa gccaacgaaa tgtgatggag atttactgaa 660

gctaagcatt aattttgggg ataggatgtt ctcgatgtag gcctagatgt atgcagcatc 720

gtgtaaggct atctattaaa gaagtctaaa aaattcactt gtgatattga gtcataaagt 780

tggtttttaa tccattctca acagactctc aattt 815

<210> 16

<211> 603

<212> DNA

<213> human (Homo sapiens)

<400> 16

tctagctcag ggcatcaaca aacagtacaa aaatgagagt gcaaatgaat aaataaatgg 60

agaatggatg aataaattaa ggaattataa aaattaagaa aaaactaatc tgtttattta 120

agtgcaaatt attttagctt aaattataat ggtgttctgg cggtgggggt cgggggttgg 180

ttctactata accaactcca cacaatatgg gccataaaac ctgccaggtt ttctgtgctg 240

tgggcatgtt cagaagaatg tatgtgactc tgttttttca gtgctatcct tttgctatca 300

caggcttcat ctatctctaa tgttgtgaca acacttgtaa agcacagtct ctgcttcttt 360

atgtagaaag tcctgggttt ttgtatcagg cagaaatttc tattaattat gagattctag 420

tatagctctg ctatacctgt ctgtatgatc cagtggcact ggtggttctc cagtgacaaa 480

gcaaagctga tgagtgttga gtatcattcc aatggaaata gtattctcat tgttgggaag 540

gcataactca agtggcatat gtaattataa tctagcaccc agcaacactt ggggaggtag 600

tgc 603

<210> 17

<211> 696

<212> DNA

<213> human (Homo sapiens)

<400> 17

aataggcaga aactggacca aagctaatca atcaaatatt actgtcttta aatgtgacca 60

tagatcttca tgtaaggaag tgaaaattca cttattcaaa tcataaacta ccaatttcca 120

aacagtaaat tctgtgagga gaaaaaaaaa aacttacata gtatcttata actgaaagaa 180

gcctcagagt cacctggtcc gtacattttg tttcagtgag gaaaatgaag gagagagggt 240

acattaacag cacataccta ttcaactatt tactgggtgg cttttacaca tcaacagaat 300

gctcttaaat atctggcagg aaaagcaatc agtgaactga tacatgatat gtcttcagag 360

tcacactcac tgctgcttac acagccacca gactgacctg atggccttag gcaagttaca 420

cttcctgctc aaaaagcttc agcctcccac tgcctccaga ataaatttta gcctgctctt 480

cagggctgtc ttccatctgg ctcaacttac ttttccatcc tttttgtcca actcgtaact 540

ccacaaaatt cbatgcttta gccaaactag aagacttgtt tttccaaaca tgccctgcag 600

tttcctcctg ccatgcaagt tgcctgtgac gttgacttgc cctgggcttc ctttccttaa 660

tgctccccgg tgcattcccc agacttcaag gctcaa 696

<210> 18

<211> 549

<212> DNA

<213> human (Homo sapiens)

<400> 18

ctacatgagg ggctattatc tccattttat agatgagaaa actaaggcac actaagatca 60

aataacttac ccaagggtat ccagcctagt aagtggctaa gctggttttg agtttaggta 120

atcaaggtcc agagtccctg ttcttaacta ccccactata ctctctctca tacagtcact 180

gaaatatgac atattttgtt gaaggggtaa atgcatgact aaattgatga atgccacatc 240

actccacttg ctaaaatatt cacatcaata agtaaatgaa tgccttctac aggctgggag 300

gagaaaaata taacaaatac aatccttgct tttaagaatg ttatagtcta gagaagtttc 360

ctcagattgg agtctgtagg tatattcttg attccattag ttcttcacaa aaagttttaa 420

cttgtatttt tattctaata gtaatttaaa ttcaaagtac atgctagatt cattttaact 480

caatactgcc attcaattct agcaaccaat attgctttgc tgtttaagat cactttggtc 540

acccaagag 549

<210> 19

<211> 706

<212> DNA

<213> human (Homo sapiens)

<400> 19

ccgggactgc aaagcaatgt gaaaaggaag caggaggggc cagacgcgcg gtttgcactg 60

gattccaggg ctgccactgc aggcgcgtgg gggagggatc ggatctgcga ggaccggggc 120

ccgccgcgcc caggcagcca cgtcgagaag ccacatagct ggggaccagg tgcacgggtg 180

cttccagcgg acttgggggt acttaccgta ctggaagtag ccagtgccgt atccgggccg 240

gtacctgccc ccaggtctgg gcctttcata agtatcgtag tagttgtaat aagggttgtc 300

gtcagagtac ttgtaggggt tgtaagggtc gtcgcccacc atgccgtcca cgcggctggg 360

cggccgcagg ttactgagcg caggaacttc tcccggcgct gtctggttct ccgcgcgcga 420

ggcgccagct tcgcgggctc tagatgtcga gtagccagct tggaaccagt gacgggcggt 480

gggcctgggg cggccagcgg tgactccaga tgagccggcc gtccgcgttc gcgccgcggc 540

ggtgcggttg tcgcggatca gcaggatcgg agtgcggggc tgctgggcgg aggcgttggc 600

tgcaccaggg acggcggcgc ccgggtcccg gcggcgctga ggctggtact gtgagcccag 660

gctcagcaag ctgaacacct gcccgttgtt ctcccattgg atctgc 706

<210> 20

<211> 505

<212> DNA

<213> human (Homo sapiens)

<400> 20

gatctgggcg ggagtcaaat tataatgtcc ttttacaaat tagggtttca tatggaaata 60

ctgacataga ttttaactga cacgcatttt ccaatgataa aacatgcaaa ctgcttagtt 120

cagcactact ttaaaaaaat ccatccaaac aacatctgac atcaattata ctgtagattt 180

aaatatatat gtgtggagaa agaaatggtg tccttctgct cttatttgca ttattaaaag 240

aggcaacttt ttaaagtgct tttaaagaaa cttatttttc ctccatttgc taaccgcaac 300

cactattcta ttttcagcat aaaacagaag gaaggaatgg tttcacaggt gaaaaaacag 360

agatatcttt ttttacagtt atttactaag ccggttaagg aatacagaat gggtgcatat 420

gttgtcaacc attcagactt tttcagagag taaatttttg ttcttcattg tggactgtaa 480

caaggaccca cactgacctg tgatc 505

<210> 21

<211> 698

<212> DNA

<213> human (Homo sapiens)

<400> 21

ctaccctttg tcacacatgc taatgaaaag cttagttgat tttcactctc cttcatttcc 60

tctaaaatct ctctggctta ggactctgtc agctgaaaaa caaacatgtg gctaccacat 120

ttggggacaa tacagtgttt gcaagccctg cgggagataa tctagccaca cattgtgttc 180

cctgttcaat aacaaaacta ttattcacaa aattggagaa accatagttc ctttccactc 240

aaatctgaga tgataatgat gatgacaata ataataataa gccacggcta catcaagata 300

caaacagctt tttttgcttt gataagatcc acagctgatt tcacttttga ccatgagatt 360

ttcttctcgt gaacaattct gcagtatgtg ccataagaga agggaaggaa tgttgctaat 420

tctttttttg agtttctagc ccatcaatat caaatcttta aatggcaatg tctggcccat 480

tggccaagaa atgaaagttg ttgtaatgct atgttcctgg tatttgttaa atacatttat 540

ttttgtagga catttcacat aaatggaaaa cagaaagccg aaaccataaa gcagggcctc 600

tgaattgcag aagccagaac agtaatgcca cattcaaagc aatcaggttc aagtgtaaat 660

tcttttgttt gaggctcaag aagctcatac aaagcttg 698

<210> 22

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 22

cgcatgatgt cctgtgtagc 20

<210> 23

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 23

tggctgttat gatacctatg gtg 23

<210> 24

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 24

catgcaacct aaatccctca t 21

<210> 25

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 25

ttgagagtct gttgagaatg ga 22

<210> 26

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 26

agctcagggc atcaacaaac 20

<210> 27

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 27

ctacctcccc aagtgttgct 20

<210> 28

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 28

aggcagaaac tggaccaaag 20

<210> 29

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 29

agccttgaag tctggggaat 20

<210> 30

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 30

catgaggggc tattatctcc a 21

<210> 31

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 31

ttgggtgacc aaagtgatct t 21

<210> 32

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 32

ggactgcaaa gcaatgtgaa 20

<210> 33

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 33

gatccaatgg gagaacaacg 20

<210> 34

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 34

ctgggcggga gtcaaattat 20

<210> 35

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 35

cacaggtcag tgtgggtcct 20

<210> 36

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 36

ccctttgtca cacatgctaa tg 22

<210> 37

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 37

gctttgtatg agcttcttga gc 22

<210> 38

<211> 573

<212> DNA

<213> human (Homo sapiens)

<400> 38

gacatggttt tgccatgttg gcccggctgg tatcaaactc ctggcctcag gtgatccacc 60

tgcctcggcc tcccaaagtg ctgggattat aggcgtgagc caccgtggct taacctaaac 120

ttctagatat gaaaactata atgtctgagg tgaaaaatag actggaggaa gaaaaaggca 180

gattggacat tgtagaagaa aggcgagatc agtgaaatta aaagtgtggc aatgaaaact 240

acccaaaata aaacacgcag aaaataaaat tccaaaaaat gaaaaagcat aagtgatatg 300

tgggatagtt tcagtggcct aatacatgag taactggagt ccctgaagaa gaaagactga 360

aaaatatatt tttagaaata atggctgaaa aatttccaaa cttagtgaag ctataaacct 420

acagatctaa gctcaatgaa caccaagaaa aataaacatg aaaataacta ccccaaatta 480

cagcataagt tactaaaaaa caatgacaaa gagaaaaatc tgacagccag aaaatatacc 540

cgtattacat ccagagaaac aaagataagg ctg 573

<210> 39

<211> 494

<212> DNA

<213> human (Homo sapiens)

<400> 39

cgtgctggca aacttgaaat ctgatgaacc cactaccttc actacactca ctgtctggtc 60

tctctcaggt agctgaggac tgagaaacac cccagccagg caggttgggg tcactataaa 120

ttcacagtca ccaaccacac atggtgctct gtttggctgg ccatctcatt atacctgtct 180

gacttgctta ctgtcctgcc ctccgaaaaa gcttttcaaa cactatctcc ttctgcctca 240

aatgtatcac ctccccctca atgactttgc ttcatatgtg acacaaagaa atgttaatac 300

cagccaagac cttcctcaac ttccggccag caaatctgga acacaagctg catctgtcct 360

ggtttgcctc ttctctcttc tcgttacagt agagtagcca ttcctcttct aaaaaacact 420

gatctcccat cctgctctgg acattaaagt actgagaagt ggttttgtgt tcccagcaat 480

atgggagtca ttgg 494

<210> 40

<211> 450

<212> DNA

<213> human (Homo sapiens)

<400> 40

aagtgatcca cccacctgag cctcccaaag ttctgggatt acaggcttga gccactgcgc 60

ccagctataa ccttcacgtt caaagcactc tcccatctat tattaaggcc tctgagaaaa 120

tctccttttt atatatattc tcaagtcaga attattagtc tccatctttc atcttagtct 180

tcttcaaata ccaaactctt tactaagcag ttagacacaa gtagtttcac cagttctatc 240

actcaaatac ggtttcataa gttctgccta gtgattgccc atcagagggg atatggattt 300

gtgggcagga aatagaatag aaacctaact ctaactcttc tgtgaaatgt agtgtttagg 360

ttaatattct tccttaactg ccatctgctt ttttttgttt gtttaacact ttgtccaaat 420

ctctcaaacc ttacaccgca gaatcatgga 450

<210> 41

<211> 568

<212> DNA

<213> human (Homo sapiens)

<400> 41

gctttcatgg tgtgggctaa ggacgagcgc aagcggctgg cgcagcagaa tccagacctg 60

cacaacgccg agttgagcaa gatgctgggt gagtccgagt cgcagaccca ggcggccggg 120

cgcgctggcg cgaatcgcta ggccgatttc ttaaacccca aactgttctt tgcgagcctg 180

acgcccaaaa ccaggggtgt gtagcggcca cgtcctttct taaggctctg ggttcccttc 240

ccgcttcccg ccctccgacc ctccaaagca gctttccgcc ttgctctccg gctcccggat 300

tccccaggtg gccgggggcg cgggtccaac ggctctggga aggcgacttc ccggcacctc 360

cgggcggcgc gagagcaccc ttggccctga actgggccgg ttgtgtccat ccctcgaccc 420

cttccctagt taggtgtcct tttctgtttt tcgaacgacc gggtgatggg tgagcggaaa 480

gccgcttcca ggagaccaaa agaaaggggt gcctttagag gacgggtgtt ccccaagggc 540

tcggactcag gagtcccaga tctccctc 568

<210> 42

<211> 529

<212> DNA

<213> human (Homo sapiens)

<400> 42

aggtgtacca accccagaca ctggcagaac gctctcttta tgtaagacct caaaatctta 60

ctggttggtg tctgctgact tgtttttctc tgttatcttg cctgtctgca gtgacaaatt 120

cagtgcagct ctaactcatg tggacaggga ggaaatgatt ctaggattga ggacttaagg 180

gtgtctggaa gagaagagaa ttgttttgtt ttgttttgtt ttgttagttg ttgttttcct 240

cagacaggac cttgtcaacg ctttcaaata tgtaggctgt ttgctgtttc ctttatgttg 300

gtccctgaga aggatctgcc cttctaccct gtttccctgg ggggtgtgga cagagccttt 360

gtctttgggg aagggggtca tcttgggaaa aggagaacag ggcatcctga ggacctgctc 420

cgtctaagga gagaaagccg aacagatggc agctgccacg cagagggcac tttgtaggaa 480

ctctggctgg agcaggcttc ctgcgctcca atgccaaatc cttcccttc 529

<210> 43

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 43

gacatggttt tgccatgttg 20

<210> 44

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 44

cagccttatc tttgtttctc tgg 23

<210> 45

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 45

cgtgctggca aacttgaaat 20

<210> 46

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 46

ccaatgactc ccatattgct g 21

<210> 47

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 47

aagtgatcca cccacctgag 20

<210> 48

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 48

tccatgattc tgcggtgtaa 20

<210> 49

<211> 19

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 49

gctttcatgg tgtgggcta 19

<210> 50

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 50

gagggagatc tgggactcct 20

<210> 51

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (Forward)

<400> 51

aggtgtacca accccagaca 20

<210> 52

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> primer (reverse)

<400> 52

gaagggaagg atttggcatt 20

Claims

1. An SNP marker composition for diagnosing cerebral aneurysm, characterized by: including rs371331393, which is a Single Nucleotide Polymorphism (SNP) of ARHGAP32(Rho GTPase Activating Protein 32) gene.

2. The SNP marker composition for diagnosing cerebral aneurysm according to claim 1, wherein: rs371331393 is an SNP of the ARHGAP32 gene, including the 26 th nucleotide of SEQ ID NO. 4.

3. The SNP marker composition for diagnosing cerebral aneurysm according to claim 1, wherein: the composition further comprises 1 or more SNPs selected from the group consisting of:

1) rs75822236, which is a SNP of the GBA (gluconerebrosidase) gene;

2) rs112859779, which is a SNP of TCF24(transcription factor 24) gene;

3) rs79134766, which is an SNP of OLFML2A (Olfactomedin Like 2A) gene;

4) rs 13852525217, which is an SNP of the CD163L1(CD163 molecular Like 1) gene;

5) rs74115822, which is a SNP of the CUL4A (cullin 4A) gene;

6) rs75861150, which is a SNP of the LOC102724084 gene;

7) rs116969723, which is a SNP of LRRC3 (leucoine rich repeat conjugation 3) gene;

8) rs6741819, which is a SNP of RNF144A (Ring Finger Protein 144A) gene;

9) rs59626274, which is a SNP of the FLJ45964 gene;

10) rs17688188, which is a SNP of SPCS3(Signal peptide Complex subBunit 3) gene;

11) rs56942085, which is a SNP of LINGO2(Leucine Rich Repeat And Ig Domain containment 2) gene; and

12) rs72835045, which is a SNP of MINK1(misshapen/Nck-interacting kinase (NIK) -related kinase 1) gene.

4. A composition for diagnosing a cerebral aneurysm, comprising: an agent of rs371331393, which is a SNP of the ARHGAP32 gene, can be detected or amplified.

5. The composition for diagnosing cerebral aneurysm according to claim 4, wherein: the composition further comprises more than 1 SNP that can be detected or amplified selected from the group consisting of:

1) rs75822236, which is a SNP of the GBA gene;

2) rs112859779, which is a SNP of the TCF24 gene;

3) rs79134766, which is an SNP of the OLFML2A gene;

4) rs 13852525217, which is a SNP of the CD163L1 gene;

5) rs74115822, which is a SNP of the CUL4A gene;

6) rs75861150, which is a SNP of the LOC102724084 gene;

7) rs116969723, which is a SNP of LRRC3 gene;

8) rs6741819, which is a SNP of RNF144A gene;

9) rs59626274, which is a SNP of the FLJ45964 gene;

10) rs17688188, which is a SNP of the SPCS3 gene;

11) rs56942085, which is a SNP of LINGO2 gene; and

12) rs72835045, which is a SNP of the MINK1 gene.

6. An information providing method for diagnosing a cerebral aneurysm, comprising: the method comprises the following steps: a step of determining an rs371331393 allele of an ARHGAP32 gene in a biological sample isolated from an individual, wherein the rs371331393 is an SNP of the ARHGAP32 gene.

7. The information providing method for diagnosing a cerebral aneurysm according to claim 6, further comprising: rs371331393 is an SNP of the ARHGAP32 gene, and when the allele is G, it is a step of diagnosing the onset of cerebral aneurysm or predicting the high risk of the invention.

8. The information providing method for diagnosing a cerebral aneurysm according to claim 6, wherein: the method further comprises the following steps: a step of determining alleles of 1 or more SNPs selected from the group consisting of:

1) rs75822236, which is a SNP of the GBA gene;

2) rs112859779, which is a SNP of the TCF24 gene;

3) rs79134766, which is an SNP of the OLFML2A gene;

4) rs138525217 is a SNP of the CD163L1 gene;

5) rs74115822, which is a SNP of the CUL4A gene;

6) rs75861150, which is a SNP of the LOC102724084 gene;

7) rs116969723, which is a SNP of LRRC3 gene;

8) rs6741819, which is a SNP of RNF144A gene;

9) rs59626274, which is a SNP of the FLJ45964 gene;

10) rs17688188, which is a SNP of the SPCS34 gene;

11) rs56942085, which is a SNP of LINGO2 gene; and

12) rs72835045, which is a SNP of the MINK1 gene.

9. The information providing method for diagnosing a cerebral aneurysm according to claim 8, wherein: the method further comprises the following steps:

1) rs75822236 is a SNP of the GBA gene, when its allele is C;

2) rs138525217 is a SNP of the CD163L1 gene, when its allele is C;

3) rs74115822 is an SNP of CUL4A gene, when its allele is G;

4) rs56942085 is SNP of LINGO2 gene, when its allele is G,

or a combination of two or more of them, a step of diagnosing the onset of a cerebral aneurysm or predicting a high risk of onset of the cerebral aneurysm.

10. The information providing method for diagnosing a cerebral aneurysm according to claim 8, wherein: the method further comprises the following steps:

1) rs112859779 is a SNP of TCF24 gene, when its allele is T;

2) rs79134766 is an SNP of OLFML2A gene, when its allele is a;

3) rs75861150 is a SNP of LOC102724084 gene, when its allele is C;

4) rs116969723 is a SNP of LRRC3 gene, when its allele is a;

5) rs6741819 is an SNP of RNF144A gene whose allele is T;

6) rs59626274 is a SNP of the FLJ45964 gene, when its allele is T;

7) rs17688188 is a SNP of the SPCS3 gene, when its allele is A;

8) rs72835045 is SNP of MINK1 gene, when its allele is A,

or a combination of two or more of them, and diagnosing the onset of cerebral aneurysm or predicting a low risk of onset of cerebral aneurysm.

11. The information providing method for diagnosing a cerebral aneurysm as set forth in claim 6, wherein: the individuals are Korean and Asians.