KR101992785B1 - Method for providing information of prediction and diagnosis of hypertension using methylation level of GNAS gene and composition therefor - Google Patents

Abstract

The invention GNAS The GNAS methylation biomarker according to the present invention can be used not only for predicting pre-hypertension or hypertension, but also for predicting hypertension or hypertension, Screening, risk assessment, diagnosis of disease, diagnosis of stage of disease, and selection of treatment targets.

Description

[0001] The present invention relates to a method for providing information for predicting or diagnosing hypertension using the level of methylation of the GNAS gene and a composition therefor,

The present invention relates to a method of providing information for the prediction or diagnosis of hypertension using a methylation level of the GNAS gene and compositions for this purpose, specifically, GNAS A method for providing information for prediction or diagnosis of hypertension using the methylation level of a promoter region of a gene, a composition and a kit for predicting or diagnosing hypertension.

Hypertension is the highest incidence of cardiovascular disease among the world, and there are 1 billion patients worldwide. In addition, hypertension is the most common type of cardiovascular disease, In Korea, 30% of adults over 30 are suffering from hypertension.

Hypertension refers to the case of a systolic blood pressure of 140 mmHg or more or a diastolic blood pressure of 90 mmHg or more. Symptoms can lead to death if serious, such as headache, dizziness, heart attack, dyspnea, sleep disorder, dysmenorrhea and arteriosclerosis. According to the data provided by the National Hypertension Center, the cause of primary hypertension (essential hypertension), which accounts for the majority of 90% of hypertension, is still uncertain, And dietary habits, and secondary hypertension accounts for 10% of all cases, which are caused by specific causes such as endocrine diseases, aortic stenosis, kidney disease, and drugs.

Essential hypertension is more common in adolescence than in younger age and has a family history. Genetic abnormalities that cause essential hypertension have not been elucidated as genetic factors, but twin or family studies have shown that 30-60% of hypertension is explained by genetic factors . The clinical significance of these genetic factors is to prevent the occurrence of hypertension complications by frequently measuring blood pressure and inhibiting exposure to environmental factors of elevated blood pressure in the presence of family history of hypertension.

Recently, methods for diagnosing diseases such as cancer through DNA methylation measurement have been suggested. When the CpG islands of a particular gene are hypermethylated, expression of the gene is silenced. This is a major mechanism in which the function of the gene is lost without mutation in the coding sequence of the gene coding sequence in the living body. In particular, the function of tumor suppressor genes in the cancer is lost It is interpreted as a cause. Therefore, searching methylation of CpG islands of a specific gene is a great help in disease research, and it has recently been actively tried to use such as methylation specific PCR or automatic base analysis for diagnosis and screening of diseases .

In the genomic DNA of mammalian cells, there is a fifth base besides A, C, G, and T, which is 5-methylcytosine (5-mC) with a methyl group attached to the 5th carbon of the cytosine ring. 5-mC always comes only to C of the CG dinucleotide (5'-mCG-3 '), and this CG is often referred to as CpG. Most of C of CpG is methylated with a methyl group. Such methylation of CpG inhibits the repetitive sequence in the genome, such as Alu and transposon, from being expressed, and is the most common site of extracellular changes in mammalian cells. The 5-mC of this CpG is deaminated naturally to T, and thus the CpG in the mammalian genome has a frequency of 1% which is much lower than the normal frequency (1/4 x 1/4 = 6.25%) .

There are exceptions in CpG that appear to be dense, and this is called CpG island. CpG islands are 0.2 to 3 kb in length, and the distribution percentage of C and G bases is more than 50% and the distribution percentage of CpG is higher than 3.75%. About 45,000 CpG islands are found in the whole human genome, and they are concentrated on promoter regions that regulate gene expression. In somatic cells of normal individuals, CpG islands are not methylated in these important gene expression regulatory regions, but imprinted genes and inactivated genes on X chromosomes are methylated so that they are not expressed during development .

Many diseases are caused by gene abnormalities, and the most common type of gene abnormalities is a change in the coding sequence of a gene, which is called a genetic change. When there is a mutation in a gene, the protein that the gene encodes changes in structure and function, leading to disability and deficiency, and this mutant protein causes disease. However, if there is an abnormality in the expression of the gene without mutation in the gene, the disease can be induced. A typical example is the methylation of methylation at the cytosine base site of the regulatory CpG island of gene transcription, in which case the gene is blocked from expression. This is called epigenetic change, which, like the mutation, is transmitted to progeny cells and causes the same effect, the loss of expression of the protein.

Although there exist some techniques for diagnosing or predicting specific cancers by analyzing the level of methylation of genes, there have been no reports of predicting or diagnosing total hypertension or hypertension through the analysis of methylation level of a specific gene. Accordingly, the present inventors developed a specific methylation marker for predicting or diagnosing an effective pre-hypertension or hypertension capable of early diagnosis or prediction.

Korean Patent Publication No. 2005-0057593 discloses a method for diagnosing hypertension risk and Korean Patent No. 1587635 discloses a method for detecting methylation of a thyroid cancer-specific methylation marker gene for diagnosis of thyroid cancer , There is no description of the 'method for providing information for prediction or diagnosis of hypertension using the methylation level of the GNAS gene and the composition thereof' of the present invention.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs. The present inventors conducted whole genome bisulfite sequencing (WGBS) from a sample of a normal group and a hypertensive group, and found that a base of SEQ ID NO: 1 The GNAS complex locus promoter region consisting of the nucleotide sequence was selected and DNA methylation sequencing was performed on the region. As a result, the hypermethylation level of the region was confirmed in the hypertensive group, thereby completing the present invention.

In order to solve the above problems, the present invention provides a method for separating GNAS Detecting methylation of the promoter region of the gene; And comparing the detected methylation to promoter site methylation of the GNAS gene of a normal control sample.

The present invention also provides a composition for predicting or diagnosing hypertension comprising a substance capable of detecting whether methylation of the GNAS gene promoter region has occurred .

The present invention also provides a kit for predicting or diagnosing hypertension comprising the composition as an active ingredient.

It is expected that the GNAS methylation biomarker according to the present invention can predict the onset of pre-hypertension or hypertension in an early stage. In addition, the GNAS gene of the present invention can be used for hypertension screening, risk assessment, diagnosis of disease, diagnosis of stage of disease, and selection of a therapeutic target, and may be useful for studying epigenetics of hypertension.

1 is a schematic diagram of the present invention.
Figure 2 shows the methylation rate of the GNAS gene promoter region (57464000-57464450) in the normal control (blue) and hypertensive (red) groups.

According to an aspect of the invention, the invention is separated from the biological sample GNAS Detecting methylation of the promoter region of the gene; And comparing the detected methylation to promoter site methylation of the GNAS gene of a normal control sample.

In a method according to an embodiment of the present invention, the hypertension is meant to include pre-hypertension and hypertension.

The GNAS The promoter of the gene is a sequence including a CpG island, and may be composed of the nucleotide sequence of SEQ ID NO: 1, but is not limited thereto. The nucleotide sequence of SEQ ID NO: UCSC Genome Browser Human The nucleotide at position 57,463,876-57,464,621 of chromosome 20 in the GRCh37 / hg19 DNA sequence.

The method for providing information for predicting or diagnosing hypertension according to the present invention may be,

(a) isolating genomic DNA from separate biological samples of a normal control and suspected individual;

(b) treating the separated genomic DNA with a reagent that differentially transforms methylated DNA and unmethylated DNA;

(c) detecting methylation of the promoter region of the GNAS gene in the genomic DNA treated with the reagent or a fragment thereof; And

(d) determining hypertension as an increase in the methylation level of the promoter region of the GNAS gene compared to the methylation of the GNAS gene promoter region of the normal control group.

The biological sample of step (a) of the present invention includes a wide range of all biological fluids obtained from individuals, and preferably includes cells suspected of being hypertensive or cells, tissues, biopsies, paraffin tissues, blood, serum, Plasma, urine, and combinations thereof, and more preferably, it may be blood, but is not limited thereto. Methods for obtaining body fluids, tissues, biopsies and the like from mammals can be performed by conventional methods known in the art.

The subject suspected of hypertension according to the present invention may be a subject having a genetic factor of hypertension due to a family history of hypertension but not diagnosed with hypertension, but is not limited thereto.

The reagent of step (b) of the present invention may be bisulfite, disulfite, hydrogen sulfite or a combination thereof, but is not limited thereto.

Unmethylated cytosine (C) bases on bisulfite treated DNA are deaminated and converted to uracil (U) bases, while methylated cytosine bases remain cytosine intact. That is, they are labeled with other bases so that they can be distinguished from each other depending on whether methylation of the cytosine base is present on DNA by bisulfite treatment.

In the step (c) of the present invention, the promoter region sequence of the GNAS gene for methylation detection may consist of the nucleotide sequence of SEQ ID NO: 1, but is not limited thereto. For detection of methylation, bisulfite, , Disulfite, or a combination thereof can also be included in the range of the promoter region sequence of the GNAS gene for methylation detection according to the present invention. The nucleotide sequence of SEQ ID NO: 2 may be a nucleotide sequence of SEQ ID NO: 1 that has been treated to convert a cytosine base to uracil or another base that is detected to be different from cytosine in the course of hybridization.

In addition, the detection of methylation in step (c) may be accomplished by a method selected from the group consisting of bisulfite sequencing, sequencing by synthesis, sequencing by ligation, methylation specific PCR, time methylation specific PCR), PCR using a methylated DNA-specific binding protein, methylated DNA-specific binding antibody, PCR using aptamer, or nucleic acid chip, but the present invention is not limited thereto.

The bisulfite sequencing method is a method for detecting a nucleic acid containing methylated CpG, comprising the steps of contacting a sample containing nucleic acid with an agent for transforming unmethylated cytosine, and contacting the sample with a CpG- Containing nucleic acid. Here, the oligonucleotide primer may be characterized by amplifying the nucleic acid without distinguishing the modified methylated and unmethylated nucleic acid. The amplified product can be sequenced using the Sanger method using a sequencing primer or the next generation sequencing (NGS) method to detect methylated nucleic acid.

Here, the next generation sequencing method may be a sequencing by synthesis method and a sequencing by ligation method. A feature of this method is that instead of making a bacterial clone, a single DNA fragment is spatially separated, amplified in situ (clonal amplification), and sequenced. At this time, it is called a massive parallel sequencing method because it reads hundreds of thousands of fragments at the same time. Basically, this is a sequencing by-synthesis method. In this method, mono or dinucleotides are sequentially attached to obtain a signal, which includes pyrosequencing, ion torrent, and Solexa methods.

NGS equipment based on sequencing by synthesis includes Roche's 454 platform, Illumina's HiSeq platform, Life Technology's Ion PGM platform, and finally Pacific BioSciences's PacBio platform. have. 454 and Ion PGM use emulsion PCR as a clonal amplification method and HiSeq use bridge amplification. The sequencing bi-synthase method sequentially attaches one nucleotide and detects the phosphate, hydrogen ion, or pre-attached fluorescence generated when the DNA is synthesized and reads the sequence. In the method of detecting the sequence, 454 uses a pyroseqeuncing method using phosphoric acid, and Ion PGM uses hydrogen ion detection. HiSeq and PacBio detect fluorescence and decode the sequence.

Sequencing by-ligation is a technique for identifying nucleotides at specific positions in a DNA sequence by a sequencing technique using DNA ligase. Unlike polymerase, most sequencing techniques do not use any polymerase, and DNA ligase does not ligate mismatched sequences. This is the SOLiD system. In this technique, two bases are read with spacing, which is repeated five times independently through primer reset, so that the accuracy is improved by reading each base twice in duplicate.

In the case of sequencing by-ligation, the dinucleotide primers corresponding to the corresponding nucleotide sequences are sequentially ligated among the denucleotide primer sets made of 16 combinations, and the combination of these ligation is finally analyzed to determine the nucleotide sequence of the corresponding DNA Is completed.

In the method for providing information for predicting or diagnosing hypertension according to an embodiment of the present invention, the increase of the methylation level of the promoter region of the GNAS gene of step (d) is 25, 28, 31 , 34, 47, 50, 72, 99, 109, 116, 125, 127, 131, 133, 140, 146, 148, 155, 157, 163, 170, 174, 176, 178, 184, 190, 210, 212 , 215, 230, 236, 244, 252, 254, 265, 279, 292, 314, 320, 326, 346, 349, 363, 365, 368, 374, 376, 381, 388, 403, 406, 408, 411 , 432, 439, 442, 444, 447, 456, 469, 472, 474, 479, 482, 485, 489, 493, 500, 510, 540, 547, 550, 560, 562, 570, 575, 579, 582 , 595, 598, 604, 620, 624, 628, 631, 644, 649, 655, 696 and 721 th cytosine (see Tables 5 to 7, will, relative to the baseline of the first position in the base sequence of the first (57,463,876) Table 5 to Table 7, the p value of zero describes a position in sequence).

The present invention also provides a composition for predicting or diagnosing hypertension comprising a substance capable of detecting whether or not the GNAS gene promoter region is methylated. The composition of the present invention contains a substance capable of detecting the methylation of the GNAS gene promoter region as an active ingredient. When the methylation of the GNAS gene promoter region is increased as compared with the normal control, hypertension can be determined. The GNAS gene promoter may be a nucleotide sequence of SEQ ID NO: 1, but is not limited thereto.

In hypertension predictive or diagnostic composition of the present invention, the GNAS gene promoter substance which is capable of detecting the methylation of whether the site is methylated GNAS gene promoter region primer set designed to amplify a fragment containing the, methylated GNAS gene promoter region A methylation-specific binding protein capable of binding to the methylated GNAS gene promoter region, a methylation-specific binding antibody or an extramammary and sequencing primer, a sequencing by-synthase primer, and a sequencing by-ligation primer But is not limited to, one or more selected.

Further, in the composition of the present invention, the GNAS detection of methylation whether the gene promoter region is a genomic DNA treated with of different modifications to the methylated DNA and unmethylated DNA reagents to detect methylation if the GNAS gene promoter region And the genomic DNA is selected from the group consisting of suspected hypertension or cells, tissues, biopsies, paraffin tissues, blood, serum, plasma, urine and combinations thereof derived from a subject to be diagnosed May be isolated from the biological sample, and may be, but is not limited to, genomic DNA, preferably separated from the blood sample.

In the composition for predicting or diagnosing hypertension according to the present invention, the methylation may be selected from the group consisting of bisulfite sequencing, sequencing by synthesis, sequencing by ligation, methylation specific PCR, But not limited to, specific PCR (real time methylation specific PCR), PCR using methylated DNA specific binding protein, methylated DNA specific binding antibody, PCR using aptamer, or nucleic acid chip .

The present invention also provides a kit for predicting or diagnosing hypertension comprising the composition according to the present invention as an active ingredient.

The kit for predicting or diagnosing hypertension according to the present invention comprises as an active ingredient a composition comprising a substance capable of detecting whether or not the GNAS gene promoter region comprising the nucleotide sequence of SEQ ID NO: 1 is methylated, DNA samples can be used to predict or diagnose hypertension by detecting methylation of the GNAS gene promoter region.

The genomic DNA sample is preferably CpG-containing. Typically, the CpG-containing nucleic acid may include DNA, but not limited thereto, DNA or a sample containing RNA and mRNA. Wherein the DNA or RNA may be single-stranded or double-stranded, or may contain a DNA-RNA hybrid.

The kit of the present invention comprises a partitioned carrier means for containing a sample, a second container containing a primer capable of amplifying the 5'-CpG-3 'base sequence region of the sample, and a second container containing a cleaved or truncated nucleic acid to amplify the methylated CpG- And a third container containing means for detecting the presence of nucleic acid that has not been detected. In one embodiment, the primers are selected from the group consisting of PCR for detecting methylation on the genome, methylation specific PCR, real time methylation specific PCR, methylated DNA specific binding protein , Primers for use in PCR, quantitative PCR, nucleic acid chip, sequencing, sequencing biosynthesis, or sequencing by-ligation using PCR, methylated DNA-specific binding antibody or extramammer. The carrier means is suitable for containing one or more containers such as bottles, tubes, and each container contains the independent components used in the method of the present invention. In the context of the present invention, those skilled in the art will readily be able to dispense the necessary formulation in a container.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

1. Sample Preparation

For the development of DNA methylation markers for prediction or diagnosis of hypertension according to the present invention, the following subjects were collected and genomic DNA was extracted from the blood samples and used for methylation analysis. Of the collected subjects, 104 subjects were selected in the normal group and 44 subjects in the hypertensive group were finally selected. Physical characteristics and blood analysis were performed on selected subjects (Table 2).

Subject Normal group People without hypertension, diabetes, hyperlipidemia, heart disease, paralysis (cognitive, non-cognitive)
Blood coagulant users,
A person with a BMI of 25 (male) or 23 (female) or less, and a BMI of 20 or more.
Excluding metabolic and paralytic diseases.
* The criteria for obesity were selected according to Korean obesity criteria (Korean Standards for Obesity), and hypertension, diabetes, and hyperlipidemia were selected based on past history data Hypertension group People without diabetes, hyperlipidemia, heart disease, paralysis (cognitive, non-cognitive)
Blood coagulant users,
BMI not adjusted.

delete

In addition, the present invention was conducted under the approval of the Korea Institute of Oriental Medicine, Korea Bioethics Review Committee, and clinical questionnaire data collection and human body collection were conducted after obtaining the subject's written consent.

2. Methylation analysis

Whole genome bisulfite sequencing (WGBS) was performed by selecting four genomic DNA samples from the normal and hypertensive experimental groups. Analysis of WGBS was performed with Macrogen (Korea). WGBS analysis was used to select regions with different methylation ratios in normal group and hypertensive group, and 104 samples in normal group and 44 samples in hypertensive group Bisulfite sequencing was performed for the site. Bysulfite sequencing was commissioned by LEE Co., Ltd. (Korea), and Illumina MiSeq. Platform. The reference sequence used in the analysis was Human GRCh37 / hg19 from the UCSC Genome Browser (http://genome.ucsc.edu/cgi-bin/hgGateway).

Example 1. Selection of analysis area through WGBS

WGBS analysis of genomic DNA samples from normal and hypertensive group showed significant differences in the level of methylation between the two groups at the promoter region of the GNAS gene (Table 4).

Example 2. GNAS  Promoter region (hg19: chr20: 57463876-57464621) Methylation analysis was performed to compare the normal group and hypertension group

All genomic DNA samples collected from the promoter region of the GNAS gene were used to confirm methylation. As a result, increased methylation compared to the normal group at 90 cytosine sites was confirmed in the hypertensive group samples as shown in Tables 5 to 7 below.

From the above results, hypermethylation of the promoter region of the GNAS gene could be predicted or diagnosed, and it was confirmed that GNAS of the present invention can be used as a specific methylation biomarker for prediction or diagnosis of hypertension .

<110> Korea Institute of Oriental Medicine          KOREA FOOD RESEARCH INSTITUTE <120> Method for providing information of prediction and diagnosis of          hypertension using methylation level of GNAS gene and composition          therefor <130> PN17362 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 746 <212> DNA <213> Homo sapiens <400> 1 aaggaggcgc acagcagggg taagcgccgc cgtcgcacac tttgcacgcc gcccagggct 60 acaccactca ccgaaaattg ggaggcaggt ctgggaggcg gggttgagcg cccaccgcct 120 tgggcgcgcc cgcgcccccc gccatcgcgg cccccgcgcc cccggcccac gcccgcgcgg 180 tcccggcacc ggcctgcacc cccagggtgc gcggcggccc atttgggtgc gtccccggtg 240 ggccgatttt tcgcgcttcc ccttcggttt ataggggccg ctgctatggg tcggtcctct 300 gaagaggctg gggcgtcatc ggggccggtt agaagctctg ctccccggcg gggacactca 360 gtcgcgtcgg caccgcggag cgggctgcgt caggtggctg gccggcgcgg cgctcccctg 420 ctctctggct ccgggctgcg gcgcggcggc tggagcgagc ccctgtcccg gcgcggggcg 480 gcggcgggcg gccggcaggc gctgccttgc gtgtgagtgc acctcactca catgtaagtc 540 ggggagcgcc ggggcctccc gcggaagtgc gagtcgcccg gcgcaggcaa gagccgccgg 600 gtccgggaca aggcaggggc gcccgggcgc cgctgccttg ccccggggcg cctccgggca 660 ccccccaatt ctctctcttt tctcttctcc attggcgtgc ccaagagcca aaccaaggag 720 cgcctgaggg taaacagact cctgcc 746 <210> 2 <211> 746 <212> DNA <213> Artificial Sequence <220> <223> Bisulfite-converted GNAS <400> 2 aaggaggygt atagtagggg taagygtygt ygtygtatat tttgtaygty gtttagggtt 60 atattattta tygaaaattg ggaggtaggt ttgggaggyg gggttgagyg tttatygttt 120 tgggygygtt ygygttttty gttatygygg ttttygygtt ttyggtttay gttygygygg 180 tttyggtaty ggtttgtatt tttagggtgy gyggyggttt atttgggtgy gttttyggtg 240 ggtygatttt tygygttttt ttttyggttt ataggggtyg ttgttatggg tyggtttttt 300 gaagaggttg gggygttaty ggggtyggtt agaagttttg tttttyggyg gggatattta 360 gtygygtygg tatygyggag ygggttgygt taggtggttg gtyggygygg ygtttttttg 420 ttttttggtt tygggttgyg gygyggyggt tggagygagt ttttgtttyg gygyggggyg 480 gyggygggyg gtyggtaggy gttgttttgy gtgtgagtgt attttattta tatgtaagty 540 ggggagygty ggggttttty gyggaagtgy gagtygttyg gygtaggtaa gagtygtygg 600 gttygggata aggtaggggy gttygggygt ygttgttttg tttyggggyg ttttygggta 660 ttttttaatt tttttttttt tttttttttt attggygtgt ttaagagtta aattaaggag 720 ygtttgaggg taaatagatt tttgtt 746

Claims (12)

Detecting the promoter site methylation of the GNAS gene consisting of the nucleotide sequence of SEQ ID NO: 1 from the isolated biological sample; And comparing said detected methylation to promoter site methylation of the GNAS gene consisting of the nucleotide sequence of SEQ ID NO: 1 of a normal control sample. The method according to claim 1,
(a) isolating genomic DNA from separate biological samples of a normal control and suspected individual;
(b) treating the separated genomic DNA with a bisulfite, disulfite, hydrogen sulfite or a combination thereof that differentially transforms methylated DNA and unmethylated DNA;
(c) detecting methylation of the promoter region of the GNAS gene consisting of the nucleotide sequence of SEQ ID NO: 1 in the genomic DNA treated with the bisulfite, disulfite, hygrosensulfite or a combination thereof or a fragment thereof; And
(d) determining the hypertension as an increase in the methylation level of the promoter region of the GNAS gene consisting of the nucleotide sequence of SEQ ID NO: 1 as compared to the methylation of the GNAS gene promoter region consisting of the nucleotide sequence of SEQ ID NO: 1 of the normal control group &Lt; / RTI &gt; wherein the information is for prediction or diagnosis of hypertension. [2] The method of claim 2, wherein the biological sample of step (a) is one selected from the group consisting of suspected hypertension or cells, tissues, biopsies, paraffin tissues, blood, serum, plasma, urine, Of the total blood pressure of the subject. delete The method according to claim 2, wherein the detection of methylation in step (c) is performed by a method selected from the group consisting of bisulfite sequencing, sequencing by synthesis, sequencing by ligation, methylation specific PCR, Characterized in that it is carried out by PCR using methylation specific PCR, PCR using methylated DNA specific binding protein, PCR using methylated DNA specific binding antibody or platamer, or nucleic acid chip method. / RTI &gt; 3. The method according to claim 1 or 2, wherein the methylation is detected by the presence or absence of nucleotide sequence change or the result of amplification with a primer capable of amplifying a fragment comprising a GNAS gene promoter region consisting of the nucleotide sequence of SEQ ID NO: &Lt; / RTI &gt; wherein the information is for prediction or diagnosis of hypertension. delete 1. A composition for predicting or diagnosing hypertension comprising a substance capable of detecting whether or not the GNAS gene promoter region consisting of the nucleotide sequence of SEQ ID NO: 1 is methylated. The method of claim 8, wherein the substance capable of detecting a methylation if the GNAS gene promoter region consisting of the nucleotide sequence of the Sequence ID No. 1 is designed to amplify a fragment containing the GNAS gene promoter region consisting of the nucleotide sequences of the methylation SEQ ID NO: 1 methylation-specific binding protein can be combined with the primer set consisting of base sequences of that GNAS gene consisting of the nucleotide sequences of the methylation SEQ ID NO: 1, the promoter region and to the hybridization probe, a methylation SEQ ID NO: 1 GNAS gene promoter region, which , A methylation-specific binding antibody or an exendin, a sequencing primer, a sequencing bi-synthase primer, and a sequencing bi-ligation primer. delete 9. The method of claim 8, wherein the methylation is selected from the group consisting of bisulfite sequencing, sequencing by synthesis, sequencing by ligation, methylation specific PCR, real time methylation specific PCR, PCR using methylated DNA-specific binding protein, PCR using methylated DNA-specific binding antibody or aptamer, or nucleic acid chip method. A kit for predicting or diagnosing hypertension comprising the composition of any one of claims 8, 9 and 11 as an active ingredient.

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