Genetic marker, metabolic marker and kit for statin drug muscle toxicity

Abstract

The invention discloses a genetic marker for statin muscular toxicity and a kit thereof, the genomic level of the genetic marker is remarkable, the genetic marker has a causal relationship with the statin muscular toxicity, the prediction accuracy of the statin muscular toxicity is high, the statin muscular toxicity can be reduced, and the medication compliance of a patient can be improved. The invention also provides a metabolite marker for statin drug muscle toxicity and a kit of the metabolite marker.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a genetic marker of statin myotoxicity, a metabolic marker of statin myotoxicity, and a kit for predicting statin myotoxicity. Background Art

[0002] Statins can lower low-density lipoprotein cholesterol (LDL-C) levels and are the first-line medication for primary and secondary prevention of atherosclerotic cardiovascular disease (ASCVD). However, statins can cause muscle adverse reactions such as muscle pain and abnormal creatine kinase. According to reports, the incidence of muscle symptoms in the Chinese population is 1.173%, which is close to the incidence in foreign populations (0.5-1.0%). Muscle pain and abnormal creatine kinase lead to poor compliance of patients taking statins. In the first 12 months of follow-up after taking statins, the average proportion of days covered (PDC) for patients with primary and secondary prevention of ASCVD was only 19%, of which 66% of patients had a PDC of less than 20%. Intermittent use of statins increases the risk of major adverse cardiovascular events.

[0003] Regarding the study of genetic markers of statin myotoxicity, although SNP sites related to statin myotoxicity have been found in genes such as SLCO1B1, TMEM9, ABCB1, CACNA1S, COQ2, COQ3, COQ5, GATM, RYR1, and EYS, the SNP sites related to statin myotoxicity have not reached the significance level at the genome level (p < 5×10 –8 ), although the SNP sites of SLCO1B1 and TMEM9 genes are significant at the genomic level, it is still unknown whether they are significant in the Chinese population. In addition, there is no large-scale genome-wide association analysis (GWAS) study that reveals the genetic markers of statin myotoxicity in the Chinese population. The mechanism of statin-induced muscle adverse reactions has not yet been fully elucidated. There is currently no study that reveals genetic markers that are causally related to statin myotoxicity.

[0004] At present, the research on metabolic markers of statin muscle toxicity has found that lipid combinations such as 15-HETE, 15-HETrE, Cer (d18:1 / 18:0), Cer (d18:1 / 22:0), 9-HODE, 5-HETE metabolite combination, 8_9-DHET, 15-HETE, 15-HETrE combination, 8_9-DHET, 15-HETE can be used as metabolic markers of statin muscle toxicity, but the metabolites detected are more, and the isomers in the lipids are more, and the detection method is more complicated. At present, there is no research to reveal the metabolic markers that have a causal relationship with statin muscle toxicity. Summary of the invention

[0005] In order to overcome the defects of the prior art, the technical problem to be solved by the present invention is to provide a genetic marker for statin myotoxicity, which has a significant genomic level and is a genetic marker that has a causal relationship with statin myotoxicity. It has a high prediction accuracy for statin myotoxicity, helps to reduce statin myotoxicity, and improves patient compliance with medication.

[0006] The technical scheme of the present invention is: a genetic marker for statin muscle toxicity, which includes one or more combinations of the following: rs188023796, rs566514102, rs190586080, rs199940336, rs148096882, rs1393208, rs1553319, rs140244938, rs74691889, rs72782677, rs201658393, rs3 73907065, rs1393209, rs117835046, rs190061190, rs557716092, rs550828998, rs78631901, rs60702 844, rs145925774, rs35833908, rs117238038, rs74127604, rs76246225, rs57930606, rs56280891, rs7 7262822, rs7636552, rs1471306, rs3094812, rs9816116, rs6442738, rs7636894, rs9883730, rs98371 78. rs9829406, NDUFS5, ACP5, GAA, HSD17B12, TTC39B, TYW1, LRRC14, HLA-DOB, HLA-L, CCDC32, RP11-195 F19.9, RP11-252K23.2, RP11-526I2.5, WAC-AS1, PRKXP1, USP32P2, ANAPC1P2, COQ5, KHK, NAGPA, DAGLB , ALDH16A1, DUS3L, FN3KRP, GSTM3, DDT, TMEM176B, MOB3C, SAMD9L, RRP12, ZNF83, MIF-AS1, ZFAND2A-DT.

[0007] The beneficial effects of the present invention are: providing genetic markers for statin myotoxicity, which are significant at the genomic level and are genetic markers that have a causal relationship with statin myotoxicity. The prediction accuracy of statin myotoxicity is high, which helps to reduce statin myotoxicity and improve patient compliance with medication.

[0008] A kit for genetic markers of statin muscle toxicity is also provided, which includes a storage medium on which genetic markers of statin muscle toxicity are stored. The statin muscle toxicity-related SNP sites are analyzed by GWAS chi-square test. When any of the following results is obtained, the risk of statin muscle toxicity is increased: rs188023796 is allele T, rs566514102 is allele G, rs190586080 is allele T, rs1393208 is allele G, rs1553319 is allele T, rs140244938 is allele A, rs74691889 is allele G, rs72782677 is allele T, and rs201658393 is allele C, rs373907065 is allele C, rs1393209 is allele G, rs190061190 is allele G, rs557716092 is allele G, rs550828998 is allele G, rs78631901 is allele T, rs60702844 is allele A, rs145925774 is allele G, rs35833908 is allele C, rs117238038 is allele T, rs74127604 is allele G, rs76246225 is allele T, rs57930606 is allele T, rs56280891 is allele G, and rs77262822 is allele T.

[0009] A kit for genetic markers of statin muscle toxicity is also provided, which includes a storage medium on which genetic markers of statin muscle toxicity are stored. Gene fine positioning is performed using the SuSiE model, and the risk of statin muscle toxicity increases when any of the following results is obtained: rs188023796 is allele T, rs7636552 is allele T, rs1471306 is allele T, rs140244938 is allele A, rs3094812 is allele A, rs145925774 is allele G, rs9816116 is allele T, rs6442738 is allele C, rs7636894 is allele T, rs9883730 is allele T, rs9837178 is allele G, and rs9829406 is allele T.

[0010] A kit for genetic markers of statin muscle toxicity is also provided, which includes a storage medium on which genetic markers of statin muscle toxicity are stored. Through Mendelian randomization analysis, when any of the following is upregulated, the risk of statin muscle toxicity increases: NDUFS5, ACP5, GAA, HSD17B12, TTC39B, TYW1, LRRC14, HLA-DOB, HLA-L, CCDC32, RP11-195F19.9, RP11-252K23.2, RP11-526I2.5, WAC-AS1, PRKXP1, USP32P2, and ANAPC1P2.

[0011] Also provided is a kit for genetic markers of statin muscle toxicity, which includes a storage medium on which genetic markers of statin muscle toxicity are stored. Through Mendelian randomization analysis, when the expression of any of the following is downregulated, the risk of statin muscle toxicity increases: COQ5, KHK, NAGPA, DAGLB, ALDH16A1, DUS3L, FN3KRP, GSTM3, DDT, TMEM176B, MOB3C, SAMD9L, RRP12, ZNF83, MIF-AS1, and ZFAND2A-DT.

[0012] Furthermore, statin myotoxicity is defined as creatine kinase greater than 200 U / L (units / liter) or symptoms of muscle pain after taking statins.

[0013] A metabolic marker of statin muscle toxicity, methylsuccinylcarnitine, is also provided.

[0014] A kit for metabolic markers of statin muscle toxicity is also provided, which includes a storage medium on which metabolic markers of statin muscle toxicity are stored. When the concentration of methylsuccinylcarnitine in plasma increases, the risk of statin muscle toxicity increases. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 Locuszoom map of SNP sites significantly associated with statin muscle toxicity. Figure 1 A- Figure 1 F shows the single nucleotide polymorphism (SNP) signals, genetic recombination rates, and gene annotations within ±1.5Mb of rs188023796, rs199940336, rs140244938, rs201658393, rs145925774, and rs77262822. Potential causal variants identified by fine mapping have been identified and annotated as follows: rs7636552, rs1471306 ( Figure 1B); rs3094812 ( Figure 1 D); rs9816116, rs6442738, rs7636894, rs9883730, rs9837178, rs9829406 ( Figure 1 F). Among them, chromosome is chromosome and recombination rate is recombination rate.

[0016] Figure 2 This is a forest plot of the causal relationship between whole blood and muscle expressed genes and statin muscle toxicity. The odds ratio (OR) and its 95% confidence interval (95%CI) of the incidence of statin-induced muscle adverse reactions when the gene transcription level increases by one standard deviation. The light black error bars represent the GTEs eQTLs V8 whole blood dataset, the gray error bars represent the GTEs eQTLs V8 skeletal muscle dataset, and the black error bars represent the eQTLsGen phase Ⅰ dataset.

[0017] Figure 3 The forest plot shows the causal relationship between plasma methylsuccinylcarnitine concentration and statin muscle toxicity. The odds ratio (OR) and its 95% confidence interval (95%CI) of the incidence of statin muscle adverse reactions when the plasma metabolite concentration increases by one standard deviation. DETAILED DESCRIPTION

[0018] The present invention provides a genetic marker for statin muscle toxicity, which comprises one or more combinations of the following: rs188023796, rs566514102, rs190586080, rs199940336, rs148096882, rs1393208, rs1553319, rs140244938, rs74691889, rs72782677, rs201658393, rs3739 07065, rs1393209, rs117835046, rs190061190, rs557716092, rs550828998, rs78631901, rs60702844 , rs145925774, rs35833908, rs117238038, rs74127604, rs76246225, rs57930606, rs56280891, rs772 62822, rs7636552, rs1471306, rs3094812, rs9816116, rs6442738, rs7636894, rs9883730, rs9837178 , rs9829406, NDUFS5, ACP5, GAA, HSD17B12, TTC39B, TYW1, LRRC14, HLA-DOB, HLA-L, CCDC32, RP11-195F 19.9, RP11-252K23.2, RP11-526I2.5, WAC-AS1, PRKXP1, USP32P2, ANAPC1P2, COQ5, KHK, NAGPA, DAGLB, ALDH16A1, DUS3L, FN3KRP, GSTM3, DDT, TMEM176B, MOB3C, SAMD9L, RRP12, ZNF83, MIF-AS1, ZFAND2A-DT.

[0019] The beneficial effects of the present invention are: providing genetic markers for statin myotoxicity, which are significant at the genomic level and are genetic markers that have a causal relationship with statin myotoxicity. The prediction accuracy of statin myotoxicity is high, which helps to reduce statin myotoxicity and improve patient compliance with medication.

[0020] A kit for genetic markers of statin muscle toxicity is also provided, which includes a storage medium on which genetic markers of statin muscle toxicity are stored. The statin muscle toxicity-related SNP sites are analyzed by GWAS chi-square test. When any of the following results is obtained, the risk of statin muscle toxicity is increased: rs188023796 is allele T, rs566514102 is allele G, rs190586080 is allele T, rs1393208 is allele G, rs1553319 is allele T, rs140244938 is allele A, rs74691889 is allele G, rs72782677 is allele T, and rs201658393 is allele C, rs373907065 is allele C, rs1393209 is allele G, rs190061190 is allele G, rs557716092 is allele G, rs550828998 is allele G, rs78631901 is allele T, rs60702844 is allele A, rs145925774 is allele G, rs35833908 is allele C, rs117238038 is allele T, rs74127604 is allele G, rs76246225 is allele T, rs57930606 is allele T, rs56280891 is allele G, and rs77262822 is allele T.

[0021] In addition, a kit for genetic markers of statin muscle toxicity is also provided, which includes a storage medium on which genetic markers of statin muscle toxicity are stored. Gene fine positioning is performed using the SuSiE model, and the risk of statin muscle toxicity increases when any of the following results is found: rs188023796 is allele T, rs7636552 is allele T, rs1471306 is allele T, rs140244938 is allele A, rs3094812 is allele A, rs145925774 is allele G, rs9816116 is allele T, rs6442738 is allele C, rs7636894 is allele T, rs9883730 is allele T, rs9837178 is allele G, and rs9829406 is allele T.

[0022] In addition, a kit for genetic markers of statin muscle toxicity is also provided, which includes a storage medium on which genetic markers of statin muscle toxicity are stored. Through Mendelian randomization analysis, when any of the following expressions is upregulated, the risk of statin muscle toxicity increases: NDUFS5, ACP5, GAA, HSD17B12, TTC39B, TYW1, LRRC14, HLA-DOB, HLA-L, CCDC32, RP11-195F19.9, RP11-252K23.2, RP11-526I2.5, WAC-AS1, PRKXP1, USP32P2, and ANAPC1P2.

[0023] In addition, a kit for genetic markers of statin muscle toxicity is also provided, which includes a storage medium on which genetic markers of statin muscle toxicity are stored. Through Mendelian randomization analysis, when the expression of any of the following is downregulated, the risk of statin muscle toxicity increases: COQ5, KHK, NAGPA, DAGLB, ALDH16A1, DUS3L, FN3KRP, GSTM3, DDT, TMEM176B, MOB3C, SAMD9L, RRP12, ZNF83, MIF-AS1, and ZFAND2A-DT.

[0024] Furthermore, statin myotoxicity is defined as creatine kinase greater than 200 U / L or muscle pain symptoms after taking statins.

[0025] A metabolic marker of statin muscle toxicity, methylsuccinylcarnitine, is also provided.

[0026] A kit for a metabolic marker of statin myotoxicity is also provided, which includes a storage medium on which a metabolic marker of statin myotoxicity is stored. When the concentration of methylsuccinylcarnitine in plasma increases, the risk of statin myotoxicity increases. Furthermore, statin myotoxicity is defined as: after taking a statin, creatine kinase is greater than 200 U / L or there are symptoms of muscle pain.

[0027] The embodiments of the present invention are described in detail below.

[0028] In the present invention, a total of 1321 patients taking statins were collected, 308 patients who did not have creatine kinase detected and did not have muscle pain were excluded, and a total of 1013 patients were included. According to the creatine kinase greater than 200 U / L or muscle pain symptoms after taking statins, the patients were divided into a muscle toxicity group of 147 cases and a control group of 866 cases.

[0029] Peripheral blood was collected from patients, and the Infinium Asian Screening Array-24+ v1.0 gene chip was used to perform genotyping of SNP (single nucleotide polymorphism) sites. Genotype interpolation was performed using Impute 2 software based on the reference genome 1000 Genomes Project (phase 3). Correlation analysis was performed using PLINK v1.9 software, with a p value of less than 5×10 –8 Defined as genomic significance. Fine-mapping was performed using the Sum of Single Effects (SuSiE) model. The susieR package was used to analyze the GWAS summary data and calculate the posterior inclusion probability (PIPs) and 95% confidence interval of the SNP sites within the range of Top SNP + / - 1.5 Mb. The Locuszoomr package was used to plot the SNP signals, annotated genes, and genetic recombination rates within the range of Top SNP + / - 1.5 Mb. Muscle and whole blood cis-eQTLs were obtained from the GTEs eQTLs V8 dataset and the eQTLsGen phase Ⅰ dataset, with p values ​​less than 5×10 –8 The significant eQTLs were used as exposure data. The results of statin muscle toxicity GWAS were used as outcome data. Two-sample Mendelian randomization was used to analyze genes that were causally related to statin muscle toxicity. Metabolite-related QTLs (mQTLs) datasets from three studies by Chen Y et al., Karjalainen MK et al., and Ottensmann L et al., with p values ​​less than 5×10 –8 The significant mQTLs were used as exposure data. The TwoSampleMR package was used to analyze the whole blood, muscle expressed genes and plasma metabolites that were causally related to statin myotoxicity. The coloc package was used to perform co-localization analysis of eQTLs, mQTLs and outcomes.

[0030] The GWAS chi-square test analyzed the SNP sites associated with statin myotoxicity and found that they were significantly associated with statin myotoxicity at the genomic level (p < 5×10 -8), 27 new SNP sites were located in 20 loci, as shown in Table 1. Among them, rs188023796 (allele T), rs566514102 (allele G), rs190586080 (allele T), rs1393209 (allele G), rs1553319 (allele T), rs140244938 (allele A), rs74691889 (allele G), rs72782677 (allele T), rs201658393 (allele C), rs373907065 (allele C), rs190586080 (allele T), rs1393209 (allele G), rs1553319 (allele T), rs140244938 (allele A), rs74691889 (allele G), rs72782677 (allele T), 92 (allele G), rs550828998 (allele G), rs78631901 (allele T), rs60702844 (allele A), rs145925774 (allele G), rs35833908 (allele C), rs117238038 (allele T), rs74127604 (allele G), rs76246225 (allele T), rs57930606 (allele T), rs56280891 (allele G), and rs77262822 (allele T) are significant risk factors for statin muscle toxicity, see Table 1.

[0031] Table 1. Significant correlation between statin-induced muscle toxicity and genomic level (p < 5 × 10 -8 ) SNP site

[0032] Locuszoom shows the SNP signal, annotated genes and genetic recombination rate within the range of Top SNP + / - 1.5 Mb, see Figure 1 By applying the SuSiE algorithm, the present invention finely mapped the GWAS signals of the 20 loci Top SNP + / - 1.5 Mb gene region and successfully identified 12 potential causal variants, as shown in Table 2. It was found that rs188023796 (allele T), rs7636552 (allele T), rs1471306 (allele T), rs140244938 (allele A), rs3094812 (allele A), rs145925774 (allele G), rs9816116 (allele T), rs6442738 (allele C), rs7636894 (allele T), rs9883730 (allele T), rs9837178 (allele G), and rs9829406 (allele T) were significant risk factors for statin muscle toxicity.

[0033] Table 2. Fine mapping of SNP loci significantly associated with statin muscle toxicity

[0034] Mendelian randomization and colocalization analysis revealed that the expression of 35 genes in muscle and plasma was causally related to statin muscle toxicity, and the eQTLs were colocalized with the sites of statin muscle adverse reactions (PP.H4>0.7), see Figure 2 The up-regulation of NDUFS5, ACP5, GAA, HSD17B12, TTC39B, TYW1, LRRC14, HLA-DOB, HLA-L, CCDC32, RP11-195F19.9, RP11-252K23.2, RP11-526I2.5, WAC-AS1, PRKXP1, USP32P2, and ANAPC1P2, and the down-regulation of COQ5, KHK, NAGPA, DAGLB, ALDH16A1, DUS3L, FN3KRP, GSTM3, DDT, TMEM176B, MOB3C, SAMD9L, RRP12, ZNF83, MIF-AS1, and ZFAND2A-DT were significant risk factors for statin muscle toxicity.

[0035] The present invention uses Mendelian randomization and colocalization analysis to find that plasma methylsuccinoylcarnitine has a causal relationship with statin myotoxicity, and an increase in plasma methylsuccinoylcarnitine concentration is a significant risk factor for statin myotoxicity. Figure 3 .

[0036] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Any simple modification, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention are still within the protection scope of the technical solution of the present invention.

Claims

1. A genetic marker for statin myotoxicity, characterized by: It includes one or more of the following combinations: rs188023796, rs566514102, rs190586080, rs199940336, rs148096882, rs1393208, rs1553319, rs140244938, rs74691889, rs72782677, rs201658393, rs373907065, rs1393209, rs 117835046, rs190061190, rs557716092, rs550828998, rs78631901, rs60702844, rs145925774, rs 35833908, rs117238038, rs74127604, rs76246225, rs57930606, rs56280891, rs77262822, rs7636 552, rs1471306, rs3094812, rs9816116, rs6442738, rs7636894, rs9883730, rs9837178, rs982940 6. NDUFS5, ACP5, GAA, HSD17B12, TTC39B, TYW1, LRRC14, HLA-DOB, HLA-L, CCDC32, RP11-195F19.9, R P11-252K23.2, RP11-526I2.5, WAC-AS1, PRKXP1, USP32P2, ANAPC1P2, COQ5, KHK, NAGPA, DAGLB, ALD H16A1, DUS3L, FN3KRP, GSTM3, DDT, TMEM176B, MOB3C, SAMD9L, RRP12, ZNF83, MIF-AS1, ZFAND2A-DT.

2. The kit for genetic markers of statin muscle toxicity according to claim 1, characterized in that: The invention comprises a storage medium, wherein the storage medium stores genetic markers of statin myotoxicity. The statin myotoxicity-related SNP sites are analyzed by GWAS chi-square test. When any of the following results is obtained, the risk of statin myotoxicity is increased: rs188023796 is allele T, rs566514102 is allele G, rs190586080 is allele T, rs1393208 is allele G, rs1553319 is allele T, rs140244938 is allele A, rs74691889 is allele G, rs72782677 is allele T, rs201658393 is allele C, rs37390706 is allele C, and rs4286076 is allele D. 5 is allele C, rs1393209 is allele G, rs190061190 is allele G, rs557716092 is allele G, rs550828998 is allele G, rs78631901 is allele T, rs60702844 is allele A, rs145925774 is allele G, rs35833908 is allele C, rs117238038 is allele T, rs74127604 is allele G, rs76246225 is allele T, rs57930606 is allele T, rs56280891 is allele G, and rs77262822 is allele T.

3. The kit for genetic markers of statin muscle toxicity according to claim 1, characterized in that: The invention comprises a storage medium on which genetic markers of statin myotoxicity are stored. Gene fine positioning is performed through the SuSiE model. When any of the following results is obtained, the risk of statin myotoxicity increases: rs188023796 is allele T, rs7636552 is allele T, rs1471306 is allele T, rs140244938 is allele A, rs3094812 is allele A, rs145925774 is allele G, rs9816116 is allele T, rs6442738 is allele C, rs7636894 is allele T, rs9883730 is allele T, rs9837178 is allele G, and rs9829406 is allele T.

4. The kit for genetic markers of statin muscle toxicity according to claim 1, characterized in that: It includes a storage medium storing genetic markers of statin muscle toxicity. Through Mendelian randomization analysis, when any of the following expressions is upregulated, the risk of statin muscle toxicity increases: NDUFS5, ACP5, GAA, HSD17B12, TTC39B, TYW1, LRRC14, HLA-DOB, HLA-L, CCDC32, RP11-195F19.9, RP11-252K23.2, RP11-526I2.5, WAC-AS1, PRKXP1, USP32P2, ANAPC1P2.

5. The kit for genetic markers of statin muscle toxicity according to claim 1, characterized in that: The invention comprises a storage medium storing genetic markers of statin muscle toxicity. Through Mendelian randomization analysis, when the expression of any of the following is downregulated, the risk of statin muscle toxicity increases: COQ5, KHK, NAGPA, DAGLB, ALDH16A1, DUS3L, FN3KRP, GSTM3, DDT, TMEM176B, MOB3C, SAMD9L, RRP12, ZNF83, MIF-AS1, and ZFAND2A-DT.

6. The kit for genetic markers of statin muscle toxicity according to any one of claims 2 to 5, characterized in that: Statin myotoxicity is defined as creatine kinase greater than 200 U / L or muscle pain symptoms after taking statins.

7. Metabolic markers of statin muscle toxicity, characterized by: It is methylsuccinylcarnitine.

8. The kit for detecting metabolic markers of statin muscle toxicity according to claim 7, characterized in that: The device comprises a storage medium on which a metabolic marker of statin myotoxicity is stored. When the concentration of methylsuccinylcarnitine in plasma rises, the risk of statin myotoxicity increases.

9. The kit for detecting metabolic markers of statin muscle toxicity according to claim 8, characterized in that: Statin myotoxicity is defined as creatine kinase greater than 200 U / L or muscle pain symptoms after taking statins.

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