KR101840843B1 - Single nucleotide polymorphism marker in gene for predicting risk of drug induced leukopenia and method for predicting risk of drug induced leukopenia using the same - Google Patents

Abstract

The present invention relates to a composition for predicting drug-induced leukopenia risk including a single nucleotide polymorphism marker in APEX1 or GNB3 gene, and a method for predicting the risk of drug induced leukopenia. The single nucleotide polymorphism marker in the APEX1 or GNB3 gene according to the present invention has a high correlation with the risk of leukopenia induced by drugs, in particular, thiopurine-based drugs, and it can be used to treat leukemia, Crohn's disease, ulcerative colitis , Or a patient susceptible to leukopenia caused by thiopurine-based drug therapy such as organ transplantation, can be predicted or diagnosed early and effectively, thereby achieving the optimal therapeutic effect by administering a suitable drug to the patient The patient-customized treatment can be performed efficiently. Furthermore, the single nucleotide polymorphic marker according to the present invention can be utilized in drug development research for the treatment of drug induced leukopenia.

Description

[0001] The present invention relates to a method for predicting the risk of drug-induced leukopenia and a method for predicting the risk of leukopenia using the single nucleotide polymorphism marker,

The present invention relates to a composition for predicting drug-induced leukopenia risk including a single nucleotide polymorphism marker in APEX1 or GNB3 gene, and a method for predicting the risk of drug induced leukopenia.

6-mercaptopurine, azathiopurine and thioguanine, which are thiopurine-based drugs, are widely used in the treatment of patients such as leukemia, Crohn's disease and ulcerative colitis. It is an immunosuppressive drug. A major problem with the use of thiopurines is that about 6% of patients develop side effects such as leukopenia or hepatotoxicity even if they start taking the prescribed dose of drug calculated according to BSA. The activity of thiopurine-based drugs is associated with thiopurine S-methyltransferase (TPMT). In this regard, mutations in the TPMT gene result in decreased TPMT enzyme activity and increased TGN metabolite The results suggest that severe bone marrow suppression may occur. Therefore, to reduce this risk, the US Food and Drug Administration recommends that TPMT enzyme activity be measured before using thiopurine. However, considering the incidence of TPMT mutation in Asian population is found to be 2 to 3%, which is significantly lower than that in Caucasian (10%), it is doubtful whether the preliminary measurement of TPMT enzyme activity in Asians is useful . In addition, previous studies have shown that the NUDT15 mutation, which has been reported as a genetic cause of thiopurine-induced leukopenia, is relatively frequent in 42.5% of Korean leukopenia patients but not in the remaining patients that are not described as NUDT15 mutations Taking into account, there is still a lack of understanding of the genetic causes of leukopenia in Asian populations.

Therefore, the present inventors conducted a study to develop a marker capable of predicting the risk of leukopenia, one of side effects caused by thiopurine drugs, and found that a specific single nucleotide polymorphism site in APEX1 or GNB3 gene and a leukopenia- The present inventors have completed the present invention.

Korean Patent Application No. 10-2014-0193130

It is an object of the present invention to provide a composition for predicting leukopenia incidence risk including a single base polymorphism marker in APEX1 or GNB3 gene.

Another object of the present invention is to provide a kit for predicting the risk of leukopenia, comprising a polynucleotide that specifically hybridizes to a polynucleotide comprising the single nucleotide polymorphic marker region.

Yet another object of the present invention is to provide a method for providing information on the prediction of the risk of leukopenia, including identifying the genotype of the single nucleotide polymorphism marker.

In order to solve the above-mentioned problems, the present invention provides a nucleic acid construct comprising 10 to 100 consecutive nucleotide polymorphisms (SNPs) comprising at least 101 nucleotides of SEQ ID NO: 1 or 101 nucleotides of SEQ ID NO: DNA sequence, or a complementary polynucleotide of the polynucleotide or its complementary polynucleotide.

The present invention also relates to a polynucleotide consisting of 10 to 100 consecutive DNA sequences comprising a single base polymorphic site of any one of the 101st nucleotide of SEQ ID NO: 1 or the 101st nucleotide of SEQ ID NO: 3, or a complementary poly And a polynucleotide that specifically hybridizes with the nucleotide sequence of the polynucleotide.

In addition, the present invention provides a method for providing information on prediction of leukopenia incidence risk, comprising identifying a genotype of the 101st nucleotide of SEQ ID NO: 1 or the 101st nucleotide of SEQ ID NO: 3 from the sample DNA.

The single nucleotide polymorphism marker in the APEX1 or GNB3 gene according to the present invention has a high correlation with the risk of leukopenia induced by drugs, in particular, thiopurine-based drugs, and it can be used to treat leukemia, Crohn's disease, ulcerative colitis , Or organ transplantation, it is possible to effectively predict or diagnose a patient who is highly susceptible to leukopenia caused by thiopurine-based drug therapy, and thereby to achieve an optimal therapeutic effect by administering an appropriate drug to the patient The patient-customized treatment can be performed efficiently. Furthermore, the single nucleotide polymorphic marker according to the present invention can be utilized in drug development research for the treatment of drug induced leukopenia.

Brief Description of the Drawings Fig. 1 is a diagram illustrating a process for selecting 185 subjects for leukemia using 6-mercaptopurine according to an embodiment of the present invention.
FIG. 2 is a graph showing a result of differentiation of leukopenia in each genotype of rs2307486, which is a single nucleotide polymorphism, when 6-mercaptopurine is administered to a patient with leukemia.

Hereinafter, the present invention will be described in more detail.

In one aspect, the present invention provides a DNA sequence comprising 10 to 100 consecutive DNA sequences comprising a single nucleotide polymorphism (SNP) site of any one or more of the 101st nucleotide of SEQ ID NO: 1 or the 101st nucleotide of SEQ ID NO: A composition for predicting the risk of leukopenia, comprising a polynucleotide or a complementary polynucleotide thereof.

In the present invention, 'polymorphism' means a case where two or more alleles exist in a single gene locus, and a 'polymorphic site' means a locus in which the allele exists. Among the polymorphic sites, a single nucleotide polymorphism (SNP) is referred to as a single nucleotide polymorphism.

In the present invention, an 'allele' refers to various types of a gene existing on the same locus of a homologous chromosome. Alleles are also used to indicate polymorphism, for example, SNPs have two kinds of bialles.

In the present invention, "SEQ ID NO. 1" or "SEQ ID NO. 3" is a polymorphic sequence including a polymorphic site. A polymorphic sequence means a sequence comprising a polymorphic site comprising a SNP in a polynucleotide sequence.

Specifically, the 101st nucleotide of SEQ ID NO: 1 exists at the locus of the APEX1 gene on 14q11.2-q12, and can be represented by rs2307486. The allelic genotype of the 101 < th > nucleotide of SEQ ID NO: 1, i.e., rs2307486, is A / G. Therefore, the 101st nucleotide of SEQ ID NO: 1 may be either A or G and therefore is referred to as "r "

The APEX1 protein (NCBI accession No. NP_542380.1) was composed of 318 amino acids represented by SEQ ID NO: 2. The amino acid sequence of SEQ ID NO: 2 is the APEX1 protein sequence when the 101st nucleotide of SEQ ID NO: 1 is A (wild type). When the 101st nucleotide of SEQ ID NO: 1 is A, the 64th amino acid of APEX1 protein is isoleucine (I) , And valine (V) for G.

In the present invention, the 101st nucleotide of SEQ ID NO: 3 exists at the locus of the GNB3 gene on 12p13, and can be represented by rs2234757. The allele genotype of the 101 < th > nucleotide of SEQ ID NO: 3, i.e. rs2234757, is C / T. Thus, the 101st nucleotide of SEQ ID NO: 3 may be C or T and therefore is described as "y "

The GNB3 protein (NCBI accession No. NP_001284500.1) was composed of 339 amino acids. The amino acid sequence of SEQ ID NO: 4 is the GNB3 protein sequence when the 101st nucleotide of SEQ ID NO: 3 is C (wild type). When the 101st nucleotide of SEQ ID NO: 3 is C, the 253rd amino acid of the GNB3 protein is the aspartic acid ), And when T is aspartic acid (D).

The polynucleotide or its complementary polynucleotide according to the present invention may be composed of at least 10, preferably 10 to 100, more preferably 20 to 80, even more preferably 40 to 60 contiguous bases But are not limited thereto.

In the present invention, leukopenia includes those induced by thiopurine-based drugs. For example, leukopenia, Crohn's disease, ulcerative colitis, or organ transplantation, thiopurine- It can be induced.

Such thiopurine-based drugs include, but are not limited to, 6-mercaptopurine, azathioprine, or thioguanine.

The fact that the two monoclonal polymorphic markers can be used in predicting the risk of leukopenia in the present invention indicates that there is a specific base at a single nucleotide polymorphism site in the group in which leukopenia is caused by side effects It is based on a high probability.

The single nucleotide polymorphism marker in the APEX1 or GNB3 gene according to the present invention has a high correlation with the risk of leukopenia induced by drugs, in particular, thiopurine-based drugs, and it can be used to treat leukemia, Crohn's disease, ulcerative colitis , Or a patient susceptible to leukopenia caused by thiopurine-based drug therapy such as organ transplantation, can be predicted or diagnosed early and effectively, thereby achieving the optimal therapeutic effect by administering a suitable drug to the patient The patient-customized treatment can be performed efficiently. Furthermore, the single nucleotide polymorphic marker according to the present invention can be utilized in drug development research for the treatment of drug induced leukopenia.

In another aspect, the present invention provides a DNA sequence comprising 10 to 100 consecutive DNA sequences comprising a single nucleotide polymorphism (SNP) site of any one or more of the 101st nucleotide of SEQ ID NO: 1 or the 101st nucleotide of SEQ ID NO: And a polynucleotide that specifically hybridizes with the polynucleotide or its complementary polynucleotide to be constructed.

A polynucleotide that specifically hybridizes with the polynucleotide or its complementary polynucleotide is an allele-specific polynucleotide. Allele-specific polynucleotides are meant to specifically hybridize to each allele. That is, hybridization refers to hybridization so that the base of the polymorphic site present in the polymorphic sequence can be specifically discriminated.

But is not limited to, a probe or a primer that specifically hybridizes with the polynucleotide or its complementary polynucleotide.

The term "probe" means a nucleic acid fragment such as RNA or DNA corresponding to a few nucleotides or several hundreds of nucleotides that can specifically bind to an mRNA, and is labeled to confirm the presence or expression level of a specific mRNA . The probe may be prepared in the form of an oligonucleotide probe, a single strand DNA probe, a double strand DNA probe, or an RNA probe. Selection of suitable probes and hybridization conditions can be appropriately selected according to techniques known in the art.

The term "primer" refers to a nucleic acid sequence having a short free 3 'hydroxyl group and capable of forming a base pair with a complementary template and having a short nucleic acid sequence serving as a starting point for template strand copying It says. Primers can initiate DNA synthesis in the presence of reagents for polymerization (i. E., DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates at appropriate buffer solutions and temperatures. The PCR conditions, the lengths of the sense and antisense primers can be appropriately selected according to techniques known in the art.

The kit may be various types of kits depending on the method of using the polynucleotide, including, for example, a PCR kit, a DNA chip kit, a microarray, and the like.

In another aspect, the present invention provides a method for providing information on prediction of the risk of leukopenia, comprising identifying a genotype of the 101st nucleotide of SEQ ID NO: 1 or the 101st nucleotide of SEQ ID NO: 3 from the sample DNA.

When the genotype of the 101st nucleotide of SEQ ID NO: 1 is G or the genotype of the 101st nucleotide of SEQ ID NO: 3 is T, the risk of leukopenia is high.

More specifically, when the allele of the 101st nucleotide of SEQ ID NO: 1 is AG or GG, the risk of leukopenia is higher than that of allelotype AA.

When the allelic genotype of the 101st nucleotide of SEQ ID NO: 3 is CT or TT, the risk of leukopenia is higher than that of allelic type CC.

The method of providing information according to the present invention may further comprise separating DNA from the biological sample to obtain the sample DNA. The DNA can be isolated using phenol / chloroform extraction, SDS extraction, CTAB separation (Cetyl Trimethyl Ammonium Bromide; Murray et al., Nuc. Res., 4321-4325, 1980) or the like commercially available in the art , And is not limited thereto. The biological sample includes all tissues such as blood, saliva, urine, skin cells, mucosal cells and hair of a subject, but is not limited thereto.

Gene sequence analysis can be performed to identify the genotype. Sequence analysis can be performed using any method known in the art, for example, by using an automatic sequencer, pyrosequencing, restriction fragment length polymorphism (PCR), PCR-SSCP allele specific oligonucleotide (ASO) hybridization method, TaqMan-PCR method, MALDI-TOF / MS method, RCA (single strand conformation polymorphism), PCR-SSO method (specific sequence oligonucleotide), PCR-SSO method and dot hybridization Known methods such as rolling circle amplification, high resolution melting (HRM), primer extension, Southern blot hybridization and dot hybridization can be used, but the present invention is not limited thereto.

Hereinafter, the present invention will be described more specifically based on examples. However, the following examples are illustrative of the present invention and are not intended to limit the present invention.

Example 1. Selection of test subjects

Of the 192 leukemia patients treated at Seoul National University Hospital in Korea, 185 patients with 6 - mercaptopurine were first screened. Patients with L3 phenotype were excluded from this study because they were treated according to Burkitt lymphoma and patients who could not be informed about clinical variables could not confirm the presence of leukopenia Seven patients were excluded from this study. All participants participating in this experiment were Koreans, and the process of selecting specific subjects is shown in FIG.

According to the treatment protocols of Chidren's Cancer Group (CCG), the initial recommended dose of 6-mercaptopurine is 75 mg / m ² . But was adjusted to show a serious drug toxicity in a significant number of patients continue treatment in that capacity at Seoul National University Hospital, the results seem to stop chemotherapy bars, the initial dose to 50mg / m ² at 75mg / m ^2. The protocol for treatment was decided on a case-by-case basis by a physician responsible for modifying or discontinuing drug doses due to adverse events. Past medical records for each patient were reviewed and information on 6-mercaptopurine, including dose, duration of treatment, and timing and extent of leukopenia, was independently assessed by physicians who did not know the genotyping results.

6-mercaptopurine was prescribed on a schedule of 6-10 cycles of one cycle of the same chemotherapy that lasted for a total of 12 weeks. In the first cycle, we started taking the prescribed dose of the drug based on the body surface area (BSA) and continued to reduce the dose of 6-mercaptopurine in response to adverse events such as leukopenia or hepatotoxicity. Respectively.

In the present invention, leukopenia is divided into two major categories. According to the evaluation criteria for abnormalities in the pediatric cancer group (CCG), the leukocyte count is 50 × ^{10 9} / L or less and the age at diagnosis is 1-9 And the remaining patients are classified as high risk patients. The normal range of ANC (Absolute Neutropil count) measured by Seoul National University Hospital was over 500. The ANC number of 500-1000 was not judged to be leukopenia, but it was considered to be the boundary of leukopenia. Patients who had multiple leukopenia episodes were recorded by measuring the number of repetitions, and the dose of 6-mercaptopurine was adjusted according to the degree of leukopenia.

Target exome sequencing was performed for 40 patients with relatively leukopenia among the final selected 178 patients as in Example 2, and repeated experiments were performed through genotyping for a total of 178 patients.

Example 2. Genetic analysis through targeted exome sequencing

Of the 178 patients selected in Example 1, 40 patients who were initially treated with less than 25% of the expected dose of leukopenia and severely leukopenia were subjected to target exome sequencing after extracting DNA from blood. Seven out of 40 target exome sequencing patients were those who developed leukopenia within the first 30 days after 6-mercaptopurine treatment. Seven leukopenia patients were treated as the experimental group, and the remaining 33 patients who developed leukopenia after 30 days Were selected as the control group and analyzed using Fisher's exact test. In the Fisher's exact test, specific single nucleotide polymorphic sites and leukopenia incidence status and odds ratio (OR) were calculated by 95% confidence intervals (CI), with a statistical significance of p <0.05.

As a result, it was found that the single nucleotide polymorphism site (rs2307486, corresponding to the 101st nucleotide of SEQ ID NO: 1) on the APEX1 gene and the single nucleotide polymorphism site (rs2234757, SEQ ID NO: 3 on the 101st nucleotide on the GNB3 gene) ) Were significantly associated with leukopenia. The results of confirming the correlation between rs2307486 and leukopenia are shown in Table 1, and the results of confirming the correlation between rs2234757 and leukopenia are shown in Table 2. Data are presented in frequency and statistically significant p values are shown in bold.

As shown in Table 1, when the 33 patients who did not develop leukopenia were set as the control group, the allele frequency of the single nucleotide polymorphism site (rs2307486) on the APEX1 gene was determined as Early Grade 4 Neutropenia) (p = 0.007, OR 8.25), respectively. In a previous study (Yang SK, Hong M, Baek J, et al .: NUDT15 confers susceptibility to thiopurine-induced leukopenia Nat Genet 46: 1017-1020, In order to confirm the relationship between the mutation of rs116855232 of the gene and the group having the corresponding mutation and the case of not having mutation, it was found that there was a statistically significant correlation with the leukocytopenia in the group without the mutation of rs116855232 of the NUDT15 gene (P = 0.007, OR 12.49). Thus, it was confirmed that even at the risk of leukopenia, which could not be confirmed by the existence of the rs116855232 mutation of the NUDT15 gene, which is a known SNP marker, the single nucleotide polymorphism site (rs2307486) on the APEX1 gene of the present invention can be identified .

As shown in Table 2 above, when the 33 patients without leukopenia were set as the control group, the allele frequency of the single nucleotide polymorphism site on the GNB3 gene (rs2234757) 4 Neutropenia), respectively (p = 0.063, OR 5.55). In particular, the significance of leukopenia and mutation was evidenced in a previous study (Yang SK, Hong M, Baek J, et al: A common missense variant in NUDT 15 confers susceptibility to thiopurine-induced leukopenia Nat Genet 46: 1017-1020, . In order to confirm the relationship between the NUDT15 gene and the rs116855232 mutation in the NUDT15 gene, it was found that there was a statistically significant correlation with the leukopenia in the group having the NUDT15 gene mutation (P = 0.05, OR = 8.84), respectively.

In addition, an analysis was performed to confirm whether the possibility of early leukopenia can be predicted by combining two single nucleotide polymorphic sites identified through the above-mentioned experiment results. To this end, it is assumed that the REF (Reference) of the genotype is a reference allele in both rs2307486 and rs2234757, and HET (Heterozygote) shows A / G genotype in rs2307486 or C / T in rs2234757 HOM (Homozygote) was assumed to have a G / G genotype in rs2307486 or a T / T genotype in rs2234757. Therefore, the dominant model was statistically compared with HET + HOM for REF and the recessive model for HOM + HET + REF. At that time, no alternate allele was observed in both SNPs of the same patient. The results are shown in Table 3.

As shown in Table 3, when we combine the effects of two single nucleotide polymorphisms, rs2307486 and rs2234757, we found that the genotype frequency was significantly increased in leukopenia-induced patients (p = 0.011, OR = 10.35). This confirms that the possibility of early leukopenia can be more accurately predicted by 6-mercaptopurine therapy when both monoclonal polymorphic sites rs2307486 and rs2234757 are identified.

Example 3. Genotyping for Genetic Analysis

In order to further verify the results of Example 2, 16 experimental groups showing leukopenia incidence within the first 30 days of 6-mercaptopurine treatment and 62 control groups without leukopenia-induced genotyping for the APEX1 gene Genotyping was performed to confirm the single nucleotide polymorphism site (rs2307486). Additional validation results for the APEX1 gene are shown in Table 4.

As shown in Table 4, when the 162 patients without leukopenia incidence were set as the control group, the allele frequency of the single nucleotide polymorphism site (rs2307486) on the APEX1 gene, as in the result of Example 2, (P = 0.02, OR = 3.15) and genotype frequencies in the dominant model (AG + GG) were also increased in leukopenia patients (p = 0.07).

In addition, we analyzed the cumulative incidence of genotype (151 AA, 26 genotype, and 1 GG genotype) in order to examine the correlation between the time of onset of first leukopenia and genotype of single nucleotide polymorphism rs2307486 genotype in 178 patients . The results are shown in Fig.

As shown in FIG. 2, leucocytosis occurred at the early stage of 6-mercaptopurine treatment when rs2307486 mutation was present on the APEX1 gene, and leukopenia was observed earlier than in the case of GG genotype. The difference between groups according to genotype was statistically significant (p = 0.001527).

(Rs2307486, corresponding to the 101st base of SEQ ID NO: 1) and the single nucleotide polymorphism site on the GNB3 gene (rs2234757, corresponding to the 101st base of SEQ ID NO: 3) And the sensitivity to the reduction was confirmed.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined in the appended claims. will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

<110> Seoul National University R & DB Foundation <120> Single nucleotide polymorphism marker in gene for predicting risk          of drug induced leukopenia and method for predicting risk of drug          induced leukopenia using the same <130> 116 <160> 4 <170> Kopatentin 2.0 <210> 1 <211> 201 <212> DNA <213> Homo sapiens <400> 1 aaagaaaaat gacaaagagg cagcaggaga gggcccagcc ctgtatgagg accccccaga 60 tcagaaaacc tcacccagtg gcaaacctgc cacactcaag rtctgctctt ggaatgtgga 120 tgggcttcga gcctggatta agaagaaagg attagatgtg agtggaattt gagggaaaga 180 gacatttttt agtattgaat g 201 <210> 2 <211> 318 <212> PRT <213> Homo sapiens <400> 2 Met Pro Lys Arg Gly Lys Lys Gly Ala Val Ala Glu Asp Gly Asp Glu   1 5 10 15 Leu Arg Thr Glu Pro Glu Ala Lys Lys Ser Lys Thr Ala Ala Lys Lys              20 25 30 Asn Asp Lys Glu Ala Ala Gly Glu Gly Pro Ala Leu Tyr Glu Asp Pro          35 40 45 Pro Asp Gln Lys Thr Ser Pro Ser Gly Lys Pro Ala Thr Leu Lys Ile      50 55 60 Cys Ser Trp Asn Val Asp Gly Leu Arg Ala Trp Ile Lys Lys Lys Gly  65 70 75 80 Leu Asp Trp Val Lys Glu Glu Ala Pro Asp Ile Leu Cys Leu Gln Glu                  85 90 95 Thr Lys Cys Ser Glu Asn Lys Leu Pro Ala Glu Leu Gln Glu Leu Pro             100 105 110 Gly Leu Ser His Gln Tyr Trp Ser Ala Pro Ser Asp Lys Glu Gly Tyr         115 120 125 Ser Gly Val Gly Leu Leu Ser Arg Gln Cys Pro Leu Lys Val Ser Tyr     130 135 140 Gly Ile Gly Asp Glu Glu His Asp Gln Glu Gly Arg Val Valle Ala 145 150 155 160 Glu Phe Asp Ser Phe Val Leu Val Thr Ala Tyr Val Pro Asn Ala Gly                 165 170 175 Arg Gly Leu Val Arg Leu Glu Tyr Arg Gln Arg Trp Asp Glu Ala Phe             180 185 190 Arg Lys Phe Leu Lys Gly Leu Ala Ser Arg Lys Pro Leu Val Leu Cys         195 200 205 Gly Asp Leu Asn Val Ala His Glu Glu Ile Asp Leu Arg Asn Pro Lys     210 215 220 Gly Asn Lys Lys Asn Ala Gly Phe Thr Pro Gln Glu Arg Gln Gly Phe 225 230 235 240 Gly Glu Leu Leu Gln Ala Val Pro Leu Ala Asp Ser Phe Arg His Leu                 245 250 255 Tyr Pro Asn Thr Pro Tyr Ala Tyr Thr Phe Trp Thr Tyr Met Met Asn             260 265 270 Ala Arg Ser Lys Asn Val Gly Trp Arg Leu Asp Tyr Phe Leu Leu Ser         275 280 285 His Ser Leu Leu Pro Ala Leu Cys Asp Ser Lys Ile Arg Ser Lys Ala     290 295 300 Leu Gly Ser Asp His Cys Pro Ile Thr Leu Tyr Leu Ala Leu 305 310 315 <210> 3 <211> 201 <212> DNA <213> Homo sapiens <400> 3 ggtctgatcc ctgacccact tgccacccgt gccctcagtt cttccccaat ggagaggcca 60 tctgcacggg ctcggatgac gcttcctgcc gcttgtttga yctgcgggca gaccaggagc 120 tgatctgctt ctcccacgag agcatcatct gcggcatcac gtccgtggcc ttctccctca 180 gtggccgcct actattcgct g 201 <210> 4 <211> 339 <212> PRT <213> Homo sapiens <400> 4 Met Gly Glu Met Glu Gln Leu Arg Gln Glu Ala Glu Gln Leu Lys Lys   1 5 10 15 Gln Ile Ala Asp Ala Arg Lys Ala Cys Ala Asp Val Thr Leu Ala Glu              20 25 30 Leu Val Ser Gly Leu Glu Val Val Gly Arg Val Gln Met Arg Thr Arg          35 40 45 Arg Thr Leu Arg Gly His Leu Ala Lys Ile Tyr Ala Met His Trp Ala      50 55 60 Thr Asp Ser Lys Leu Leu Val Ser Ala Ser Gln Asp Gly Lys Leu Ile  65 70 75 80 Val Trp Asp Ser Tyr Thr Thr Asn Lys Val His Ala Ile Pro Leu Arg                  85 90 95 Ser Ser Trp Val Met Thr Cys Ala Tyr Ala Pro Ser Gly Asn Phe Val             100 105 110 Ala Cys Gly Gly Leu Asp Asn Met Cys Ser Ile Tyr Asn Leu Lys Ser         115 120 125 Arg Glu Gly Asn Val Lys Val Ser Arg Glu Leu Ser Ala His Thr Gly     130 135 140 Tyr Leu Ser Cys Cys Arg Phe Leu Asp Asp Asn Asn Ile Val Thr Ser 145 150 155 160 Ser Gly Asp Thr Thr Ala Leu Trp Asp Ile Glu Thr Gly Gln Gln Lys                 165 170 175 Thr Val Phe Val Gly His Thr Gly Asp Cys Met Ser Leu Ala Val Ser             180 185 190 Pro Asp Phe Asn Leu Phe Ile Ser Gly Ala Cys Asp Ala Ser Ala Lys         195 200 205 Leu Trp Asp Val Arg Glu Gly Thr Cys Arg Gln Thr Phe Thr Gly His     210 215 220 Glu Ser Asp Ile Asn Ale Ile Cys Phe Phe Pro Asn Gly Glu Ala Ile 225 230 235 240 Cys Thr Gly Ser Asp Ala Ser Cys Arg Leu Phe Asp Leu Arg Ala                 245 250 255 Asp Gln Glu Leu Ile Cys Phe Ser His Glu Ser Ile Ile Cys Gly Ile             260 265 270 Thr Ser Val Ala Phe Ser Leu Ser Gly Arg Leu Leu Phe Ala Gly Tyr         275 280 285 Asp Asp Phe Asn Cys Asn Val Trp Asp Ser Met Lys Ser Glu Arg Val     290 295 300 Gly Ile Leu Ser Gly His Asp Asn Arg Val Ser Cys Leu Gly Val Thr 305 310 315 320 Ala Asp Gly Met Ala Val Ala Thr Gly Ser Trp Asp Ser Phe Leu Lys                 325 330 335 Ile Trp Asn            

Claims (13)

A polynucleotide consisting of 10 to 100 consecutive DNA sequences comprising a single nucleotide polymorphism (SNP) site that is the 101st nucleotide of SEQ ID NO: 1 or a complementary polynucleotide thereof; And
A polynucleotide consisting of 10 to 100 consecutive DNA sequences, or a complementary polynucleotide thereof, comprising a single nucleotide polymorphic site that is the 101st nucleotide of SEQ ID NO: 3.
The composition according to claim 1, wherein the 101st nucleotide of SEQ ID NO: 1 is located at the locus of the APEX1 gene.
2. The composition according to claim 1, wherein the 101st nucleotide of SEQ ID NO: 3 is located at the locus of the GNB3 gene.
The composition according to claim 1, wherein the leukopenia is induced by a thiopurine-based drug.
5. The pharmaceutical composition according to claim 4, wherein the thiopurine-based drug is at least one selected from the group consisting of 6-mercaptopurine, azathioprine, and thioguanine. Compositions for predicting onset risk.
A polynucleotide consisting of 10 to 100 consecutive DNA sequences comprising a single nucleotide polymorphism (SNP) site that is the 101st nucleotide of SEQ ID NO: 1 or a polynucleotide that specifically hybridizes with its complementary polynucleotide ; And
A polynucleotide that specifically hybridizes with a polynucleotide consisting of 10 to 100 consecutive DNA sequences or a complementary polynucleotide thereof, including a single base polymorphic site that is the 101st nucleotide of SEQ ID NO: 3. Onset risk prediction kit.
7. The kit according to claim 6, wherein the specifically hybridizing polynucleotide is a probe or a primer.
Identifying the genotype of the 101st nucleotide of SEQ ID NO: 1 and the 101st nucleotide of SEQ ID NO: 3 from the sample DNA.
9. The method according to claim 8, wherein when the genotype of the 101st nucleotide of SEQ ID NO: 1 is G or the genotype of the 101st nucleotide of SEQ ID NO: 3 is T, the method predicts that the risk of leukopenia is high , Information on how to predict leukopenia risk.
9. The method according to claim 8, wherein the method comprises predicting that a risk of leukopenia is high when the allelic genotype of the 101st nucleotide of SEQ ID NO: 1 is AG or GG. Way.
9. The method according to claim 8, wherein the method comprises predicting that a risk of leukopenia is high when the allele of the 101 &lt; th &gt; nucleotide of SEQ ID NO: 3 is CT or TT. Way.
The method according to claim 8, wherein the leukopenia is induced by a thiopurine-based drug.
13. The method according to claim 12, wherein the thiopurine-based drug is at least one selected from the group consisting of 6-mercaptopurine, azathioprine, and thioguanine. How to provide information on the risk prediction.

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