氯吡格雷抗血小板效應(yīng)及藥動(dòng)學(xué)的全基因組關(guān)聯(lián)性研究
發(fā)布時(shí)間：2018-10-19 09:31
【摘要】：氯吡格雷是冠心病治療的一線抗血小板藥物,能有效降低經(jīng)皮冠狀動(dòng)脈介入治療(PCI)后冠心病患者的死亡率和再缺血事件的發(fā)生。但其療效存在較大個(gè)體差異,而目前大部分個(gè)體差異仍然無(wú)法解釋。氯吡格雷需在體內(nèi)代謝轉(zhuǎn)化為活性代謝物H4才能發(fā)揮抗血小板聚集作用,所以涉及其代謝活化的基因多態(tài)性可能對(duì)不同個(gè)體間療效的差異有重要影響。因此,本研究擬結(jié)合抗血小板效應(yīng)和藥動(dòng)學(xué)過(guò)程進(jìn)行全基因組關(guān)聯(lián)性分析(GWAS),系統(tǒng)發(fā)現(xiàn)影響中國(guó)冠心病患者氯吡格雷療效的新功能性基因變異,并評(píng)價(jià)其對(duì)主要不良心血管事件(MACE)發(fā)生的預(yù)測(cè)作用。首先,在115名患者中檢測(cè)服藥后血小板集聚率(PRU),進(jìn)行GWAS分析。同時(shí)考察H4血藥濃度及特別關(guān)注藥動(dòng)學(xué)通路的基因,篩選出18個(gè)可能影響氯吡格雷抗血小板效應(yīng)和藥動(dòng)學(xué)的候選SNP位點(diǎn)。進(jìn)一步研究候選SNP位點(diǎn)與氯吡格雷藥動(dòng)學(xué)的相關(guān)性。入選31名患者作為研究對(duì)象,給予氯吡格雷后進(jìn)行藥動(dòng)學(xué)考察及候選位點(diǎn)分型檢測(cè),分析候選位點(diǎn)基因型與氯吡格雷和H4藥動(dòng)學(xué)參數(shù)(Cmax、Tmax、AUC0-4h)的關(guān)聯(lián)性。除了CYP2C19*2外,N6AMT1 rs2254638、SLC14A2 rs12456693和ABC 1 rs2487032也與氯吡格雷的體內(nèi)藥動(dòng)學(xué)過(guò)程顯著相關(guān)。接著考察候選位點(diǎn)是否對(duì)氯吡格雷的活化代謝有潛在影響。收集32名患者正常肝組織,制備肝S9組分。建立氯吡格雷在人肝S9的體外孵育體系,分析在各個(gè)體S9中代謝物的生成量。檢測(cè)候選位點(diǎn)基因分型,分析不同基因型對(duì)氯吡格雷代謝的影響。發(fā)現(xiàn)CYP2C19*2及N6AMT1rs2254638與H4的生成顯著相關(guān)。結(jié)合上述結(jié)果發(fā)現(xiàn),N6AMT1 rs2254638和ABCA1 rs2487032是PRU的獨(dú)立影響因素,可極大提高對(duì)PRU變異的解釋程度至37.7%,而目前文獻(xiàn)報(bào)道只有約20%;N6AMT1 rs2254638和SLC14A2rs12456693是H4血藥濃度的獨(dú)立預(yù)測(cè)因子。最后,評(píng)價(jià)候選位點(diǎn)對(duì)接受氯吡格雷治療的冠心病患者M(jìn)ACE事件發(fā)生的預(yù)測(cè)作用,探尋基因多態(tài)性是否可作為與終點(diǎn)事件相關(guān)的遺傳標(biāo)記。入選患者為前瞻性研究隊(duì)列,并進(jìn)行定期隨訪。將PCI術(shù)后1.5年內(nèi)發(fā)生MACE事件的91名患者,作為病例組;另外隨機(jī)抽取208名患者,作為對(duì)照組。檢測(cè)候選位點(diǎn)分型,運(yùn)用Logistic回歸模型分析,發(fā)現(xiàn)N6AMT1 rs2254638、SLC14A2 rs12456693和ATP10Ars12913988可增加MACE事件發(fā)生的風(fēng)險(xiǎn)。推測(cè)位于N6AMT1和SLC14A2的位點(diǎn)是通過(guò)影響氯毗格雷及H4在體內(nèi)的藥動(dòng)學(xué)過(guò)程從而影響其抗血小板效應(yīng)。綜上所述,本研究系統(tǒng)發(fā)現(xiàn)了新的影響氯吡格雷療效的基因變異,極大提高對(duì)其反應(yīng)性差異的解釋程度,以及影響其療效的吸收、代謝機(jī)制的理解;有助于氯吡格雷治療的個(gè)體化用藥。
[Abstract]:Clopidogrel is a first-line antiplatelet drug for coronary heart disease, which can effectively reduce the mortality and reischemic events in patients with coronary heart disease after percutaneous coronary intervention (PCI). However, there are large individual differences in the efficacy, and most of the individual differences are still unexplained. Clopidogrel needs to be metabolized into active metabolite H4 in vivo in order to play an anti-platelet aggregation effect, so the gene polymorphism involved in its metabolic activation may have an important effect on the difference of efficacy among individuals. Therefore, this study was designed to combine the antiplatelet effect and pharmacokinetic process to analyze the genome-wide association of Chinese coronary heart disease patients with clopidogrel by (GWAS), system and to identify new functional gene mutations that affect the efficacy of clopidogrel in Chinese patients with coronary heart disease. To evaluate its predictive effect on the occurrence of major adverse cardiovascular events (MACE). First, the platelet aggregation rate (PRU),) was analyzed by GWAS in 115 patients. At the same time, the blood concentration of H4 and the genes of pharmacokinetic pathway were studied. Eighteen candidate SNP sites were screened out which may affect the antiplatelet effect and pharmacokinetics of clopidogrel. To further study the correlation between candidate SNP sites and clopidogrel pharmacokinetics. The pharmacokinetics and candidate locus typing of clopidogrel were investigated in 31 patients. The correlation between candidate locus genotypes and clopidogrel and H4 pharmacokinetic parameters (Cmax,Tmax,AUC0-4h) was analyzed. In addition to CYP2C19*2, N6AMT1 rs2254638,SLC14A2 rs12456693 and ABC 1 rs2487032 were also significantly associated with clopidogrel's pharmacokinetics in vivo. Then we investigated whether the candidate sites had a potential effect on the activation and metabolism of clopidogrel. The normal liver tissues of 32 patients were collected and liver S 9 components were prepared. The in vitro incubation system of clopidogrel in human liver S9 was established and the amount of metabolites produced in each individual S9 was analyzed. The genotypes of candidate loci were detected and the effects of different genotypes on clopidogrel metabolism were analyzed. It was found that CYP2C19*2 and N6AMT1rs2254638 were significantly correlated with H4 production. Combined with the above results, it was found that N6AMT1 rs2254638 and ABCA1 rs2487032 were independent influencing factors of PRU and could greatly improve the degree of interpretation of PRU variation to 37.70.AMT 1 rs2254638 and SLC14A2rs12456693 were only reported to be independent predictors of serum concentration of H4 by about 20% N6AMT1 rs2254638 and SLC14A2rs12456693. Finally, we evaluated the predictive effect of candidate sites on the occurrence of MACE events in patients with coronary heart disease treated with clopidogrel, and explored whether gene polymorphism could be used as a genetic marker associated with endpoint events. The patients were enrolled in a prospective study cohort and were followed up regularly. 91 patients with MACE events within 1.5 years after PCI were selected as case group, and another 208 patients were randomly selected as control group. By using Logistic regression model, we found that N6AMT1 rs2254638,SLC14A2 rs12456693 and ATP10Ars12913988 can increase the risk of MACE events. It is assumed that the sites located in N6AMT1 and SLC14A2 affect the antiplatelet effect by affecting the pharmacokinetic process of clopidogrel and H4 in vivo. To sum up, we found new gene variations that affect the efficacy of clopidogrel, greatly improve the degree of explanation of the difference in its reactivity, and influence the absorption and metabolic mechanism of clopidogrel. It is helpful for the treatment of clopidogrel.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R969
本文編號(hào)：2280746
[Abstract]:Clopidogrel is a first-line antiplatelet drug for coronary heart disease, which can effectively reduce the mortality and reischemic events in patients with coronary heart disease after percutaneous coronary intervention (PCI). However, there are large individual differences in the efficacy, and most of the individual differences are still unexplained. Clopidogrel needs to be metabolized into active metabolite H4 in vivo in order to play an anti-platelet aggregation effect, so the gene polymorphism involved in its metabolic activation may have an important effect on the difference of efficacy among individuals. Therefore, this study was designed to combine the antiplatelet effect and pharmacokinetic process to analyze the genome-wide association of Chinese coronary heart disease patients with clopidogrel by (GWAS), system and to identify new functional gene mutations that affect the efficacy of clopidogrel in Chinese patients with coronary heart disease. To evaluate its predictive effect on the occurrence of major adverse cardiovascular events (MACE). First, the platelet aggregation rate (PRU),) was analyzed by GWAS in 115 patients. At the same time, the blood concentration of H4 and the genes of pharmacokinetic pathway were studied. Eighteen candidate SNP sites were screened out which may affect the antiplatelet effect and pharmacokinetics of clopidogrel. To further study the correlation between candidate SNP sites and clopidogrel pharmacokinetics. The pharmacokinetics and candidate locus typing of clopidogrel were investigated in 31 patients. The correlation between candidate locus genotypes and clopidogrel and H4 pharmacokinetic parameters (Cmax,Tmax,AUC0-4h) was analyzed. In addition to CYP2C19*2, N6AMT1 rs2254638,SLC14A2 rs12456693 and ABC 1 rs2487032 were also significantly associated with clopidogrel's pharmacokinetics in vivo. Then we investigated whether the candidate sites had a potential effect on the activation and metabolism of clopidogrel. The normal liver tissues of 32 patients were collected and liver S 9 components were prepared. The in vitro incubation system of clopidogrel in human liver S9 was established and the amount of metabolites produced in each individual S9 was analyzed. The genotypes of candidate loci were detected and the effects of different genotypes on clopidogrel metabolism were analyzed. It was found that CYP2C19*2 and N6AMT1rs2254638 were significantly correlated with H4 production. Combined with the above results, it was found that N6AMT1 rs2254638 and ABCA1 rs2487032 were independent influencing factors of PRU and could greatly improve the degree of interpretation of PRU variation to 37.70.AMT 1 rs2254638 and SLC14A2rs12456693 were only reported to be independent predictors of serum concentration of H4 by about 20% N6AMT1 rs2254638 and SLC14A2rs12456693. Finally, we evaluated the predictive effect of candidate sites on the occurrence of MACE events in patients with coronary heart disease treated with clopidogrel, and explored whether gene polymorphism could be used as a genetic marker associated with endpoint events. The patients were enrolled in a prospective study cohort and were followed up regularly. 91 patients with MACE events within 1.5 years after PCI were selected as case group, and another 208 patients were randomly selected as control group. By using Logistic regression model, we found that N6AMT1 rs2254638,SLC14A2 rs12456693 and ATP10Ars12913988 can increase the risk of MACE events. It is assumed that the sites located in N6AMT1 and SLC14A2 affect the antiplatelet effect by affecting the pharmacokinetic process of clopidogrel and H4 in vivo. To sum up, we found new gene variations that affect the efficacy of clopidogrel, greatly improve the degree of explanation of the difference in its reactivity, and influence the absorption and metabolic mechanism of clopidogrel. It is helpful for the treatment of clopidogrel.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R969
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
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2 康彥紅;勞海燕;余細(xì)勇;陳紀(jì)言;鐘詩(shī)龍;;經(jīng)皮冠狀動(dòng)脈介入治療術(shù)后支架內(nèi)再狹窄的遺傳學(xué)與表觀遺傳學(xué)研究進(jìn)展[J];中華醫(yī)學(xué)遺傳學(xué)雜志;2012年01期
，本文編號(hào)：2280746
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