基质辅助激光解吸电离飞行时间质谱技术检测华法林代谢酶基因多态性

目的用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术建立检测人群中华法林代谢酶基因多态性的方法。方法选取2011年1月至2013年1月北京安贞医院心脏外科心脏机械瓣膜置换术后接受华法林抗凝治疗的患者10例,采集外周血,提取全血基因组DNA,用MALDI-TOF-MS技术检测10例患者华法林代谢酶基因CYP2C9*2(rs1799853)、CYP2C9*3(rs1057910)、CYP2C9*6(rs9332131)、VKORC1 CT(rs9923231)(-1639GA)、GGCX GC(rs11676382)的单核苷酸多态性(SNP)位点,并用Sanger测序法进行验证。结果用MALDI-TOF-MS技术可以同时完成10份标本3个华法林代谢酶基因5个SNP位点的检测。10名服华法林的心脏机械瓣膜置换术患者中,发现VKORC1-1639GA AG型2例,AA型8例,A等位基因频率为90%。未发现CYP2C9*2(rs1799853)、CYP2C9*3(rs1057910)、CYP2C9*6(rs9332131)和GGCX GC(rs11676382)突变。经与Sanger测序法比较,符合率为100%。结论成功建立MALDI-TOF-MS技术检测华法林代谢酶基因多态性的平台,该法具有高通量、快速、准确的特点,在个性化医疗上有较大的推广应用价值。     

华法林药物遗传学基因CYP2C9和VKORC1单核苷酸多态性在广东汉族人群的分布研究

目的探讨细胞色素P450(cytochrome P450, CYP450)家族中CYP2C9的单核苷酸多态性(single nucleotide polymorphisms, SNP)位点rs1057910和rs1799853以及维生素K环氧化物还原酶复合物亚基1(vitamin K epoxide reductase complex subunit 1, VKORC1)的rs9923231 SNP位点在广东汉族的分布,为华法林(warfarin)个体化用药(personalized medicine)提供循证医学依据.方法利用Sequenom MassArray? iPLEX GOLD系统对215例广东地区人群的rs1057910,rs1799853和rs9923231三个位点进行基因分型,并分析各基因型分布频率.结果 rs1799853和rs9923231在广东汉族中无多态性分布,分别只有CC基因型和CT基因型.rs1057910在广东汉族中有两种基因型AA和AC,分布频率分别为0.921和0.079;等位基因A的频率为0.9605,C的频率为0.0395.结论在本研究中由于rs1799853和rs9923231在广东汉族中无多态性分布,基于本研究数据建议广东汉族华法林药物的检测位点应以rs1057910为主.     

CYP2C19基因多态性与心力衰竭合并肺动脉高压的关联研究

目的探究CYP2C19 rs12248560单核苷酸多态性与心力衰竭合并肺动脉高压发生的关系。方法应用聚合酶链反应检测CYP2C19 rs12248560基因多态性,比较心力衰竭合并肺动脉高压患者组与健康人群CYP2C19 rs12248560基因型的分布频率差异以及等位基因频率差异。结果 50例心力衰竭合并肺动脉高压患者组和50例健康人群对照组CYP2C19rs12248560基因型分布频率野生型CC为52/68%,杂合型CT为28/24%,突变型TT为20/8%,两组比较χ2=7.15,P=0.027,等位基因频率野生型C为65/76%,突变型T为35/24%,两组比较χ2=5.36,P=0.024。结论心力衰竭合并肺动脉高压患者组与健康人群对照组CYP2C19 rs12248560基因型分布频率和等位基因频率有显著差异,提示心力衰竭合并肺动脉高压的发生可能与CYP2C19 rs12248560基因型多态性具有某种关联。     

1043例汉族双相情感障碍患者CYP2C19基因多态性及代谢表型分析

目的探讨汉族双相情感障碍患者CYP2C19基因多态性及代谢表型的分布,为临床合理用药提供理论参考和指导。方法以DNA微阵列技术检测1 043例双相情感障碍患者的CYP2C19基因多态性及代谢表型(快代谢型,中等代谢型,慢代谢型),用基因计数法计算基因型频率和等位基因频率,同时分析基因多态性发生频率与性别的相关性。结果所有检测对象中,CYP2C19*1/*1、*1/*2、*1/*3、*2/*2、*2/*3、*3/*3基因型的频率分别为40.6%、40.5%、5.2%、9.9%、3.4%、0.5%;CYP2C19基因快代谢型、中等代谢型、慢代谢型3种代谢表型所占比例分别为40.5%、45.8%、13.7%;不同性别之间CYP2C19基因型及代谢表型差异无统计学意义(P0.05)。结论汉族双相情感障碍人群CYP2C19基因多态性以*1/*1及*1/*2为主,681位点的变异率高于636位点,检测CYP2C19基因多态性及代谢表型有助于临床合理用药。     

华法林相关基因SNPs在心脏瓣膜置换汉族患者中的分布调查

目的:探索心脏瓣膜置换的汉族患者中华法林相关基因SNPs的分布。方法:入选2012年至2013年上海市在中山医院行心脏瓣膜置换手术后,需长期服用华法林进行抗凝治疗的汉族患者110例,通过阅读文献筛选得到华法林相关基因及相应SNPs位点(CYP2C9 rs1799853 C/T、CYP2C9rs1057910 A/C、VKORC1 rs9923231 G/A、VKORC1 rs9934438 C/T、CYP4F2 rs2108622 C/T、GGCX rs12714145 A/G、EPHX1 rs1877724 C/T、EPHX1 rs2292566 A/G、EPHX1 rs4653436 A/G)。使用直接测序法检测SNPs,采用Hardy-Weinberg遗传平衡检验验证群体代表性。结果:CYP2C9基因rs1799853、rs1057910等位基因频率分别为0.45%、5.00%;VKORC1基因rs9923231、rs9934438等位基因频率均为91.36%;CYP4F2基因rs2108622等位基因频率为28.64%;GGCX基因rs12714145等位基因频率为33.18%;EPHX1基因rs1877724、rs2292566和rs4653436等位基因频率分别为30.00%、24.55%、26.36%。结论:探索华法林相关基因CYP2C9、VKORC1、CYP4F2、GGCX、EPHX1的9个SNPs的频率在行心脏瓣膜置换术的患者人群中的分布,对建立和实施华法林个体化治疗具有重要意义。     

西北地区530例临床患者CYP2C19遗传多态性分析

目的采用DNA微阵列芯片法对西北地区人群等位基因CYP2C19*2和*3进行基因多态性检测,以了解本地区人群CYP2C19基因型。方法在完成PCR扩增获得目标片段后,运用DNA微阵列芯片法,通过具有位点特异性的寡核苷酸探针与目标序列的杂交及显色,判读基因多态性类型。结果在530份样本中,由CYP2C19*2和*3等位基因突变而产生的六种基因型(*1/*1,*2/*2,*3/*3,*1/*2,*1/*3,*2/*3)均通过DNA微阵列芯片法检测出。西北地区人群中,CYP2C19*2突变常见,为52.64%(n=279),其中,纯合突变率8.87%(n=47);CYP2C19*3基因突变率为10.94%(n=58),而该位点的纯合突变罕见,仅0.19%(n=1);CYP2C19*2/*3双突变率为3.21%(n=17)。进而据基因型判断强代谢型(EM)465例(n=210+215+40),占87.74%,弱代谢型(PM)65例(n=47+1+17),占12.26%。结论本地区人群CYP2C19快代谢与慢代谢基因型比率与国内其他地区报道相近,但发生于*2和*3位点的杂合型突变率显著高于之前文献报道。其中*2位点的突变频率较*3更为常见(52.64%vs.10.94%)。提示CYP2C19基因多态性存在一定的地区差异,掌握本地区人群的基因分布频率对临床用药具有重要的指导意义。     

四川地区冠心病患者CYP2C19基因多态性检测

目的检测四川地区冠心病患者CYP2C19基因的多态性,分析基因型与性别、年龄及临床分型之间的相关性。方法采用DNA微阵列芯片法检测131例被确诊为冠心病患者的CYP2C19基因型,结合患者的性别、年龄、临床分型进行分组,探讨这些因素与CYP2C19基因多态性的相关性。结果 131例冠心病患者中,有5种基因型被检测到,分别为CYP2C19*1/*1(636GG,681GG)、CYP2C19*1/*2(636GG,681GA)、CYP2C19*1/*3(636GA,681GG)、CYP2C19*2/*2(636GG,681AA)、CYP2C19*2/*3(636GA,681GA)。分布频率分别为53例(40.5%)、54例(41.2%)、6例(4.6%)、13例(9.9%)、5例(3.8%),而CYP2C19*3/*3(636AA,681GG)未被检测到。不同性别冠心病患者CYP2C19的基因型、等位基因频率差异无统计学意义(P0.05);不同性别的代谢型患者,其CYP2C19的基因型分布差异具有统计学意义(χ~2=6.527,P=0.038)。不同年龄和临床分型冠心病患者CYP2C19的基因型、等位基因频率、代谢型分布差异无统计学意义(P0.05)。结论冠心病患者的CYP2C19基因型分布、等位基因频率分布与性别、年龄和疾病临床分型无关。     

陕西地区CYP2C19基因多态性在不同性别、年龄、民族冠心病患者中的分布特征以及与国内其他地区的对比分析

目的探讨陕西地区冠心病治疗患者CYP2C19基因多态性分布情况。方法选取该院冠心病治疗患者4248例,检测CYP2C19的基因型,统计等位基因及代谢表型分布特征,并对比不同地区汉族和其他民族等位基因及代谢表型分布差异。结果CYP2C19*1、CYP2C19*2和CYP2C19*33种等位基因的频率分别为64.67%、30.65%、4.68%;快代谢型(*1/*1)1749例,发生率41.17%,中间代谢型(*1/*2、*1/*3)1996例,发生率46.99%,慢代谢型(*2/*2、*2/*3、*3/*3)503例,发生率11.84%;不同性别之间基因型及代谢型分布差异无统计学意义(P>0.05);不同年龄段患者CYP2C19基因型分布差异无统计学意义(P>0.05),等位基因分布差异有统计学意义(P<0.05);我国不同地区汉族人群等位基因分布差异有统计学意义(P<0.05),代谢型分布差异无统计学意义(P>0.05);不同民族等位基因及代谢型分布差异均有统计学意义(P<0.05)。结论陕西地区冠心病患者基因型主要以CYP2C19*1/*2为主,代谢表型主要以中间代谢型为主,可以评估其氯吡格雷抵抗风险,为患者制定个体化的抗血小板治疗方案。     

CYP2C19基因多态性与急性非静脉曲张性上消化道出血患者预后的关系

目的探讨CYP2C19基因多态性与急性非静脉曲张性上消化道出血(ANVUGIB)患者预后的关系。方法选取2019年4月至2020年4月该院收治的114例ANVUGIB患者为研究对象,根据预后情况将患者分为死亡组44例和存活组70例。采用实时荧光定量PCR检测CYP2C19基因多态性。比较两组患者临床资料、CYP2C19基因检测结果。采用多因素Logistic回归分析ANVUGIB患者死亡的危险因素。采用受试者工作特征曲线分析CYP2C19基因多态性对ANVUGIB患者死亡风险的预测价值。结果与存活组比较,死亡组CYP2C19*1/*1基因型、CYP2C19*1等位基因频率更低,CYP2C19*1/*2基因型、CYP2C19*1/*3基因型及CYP2C19*2等位基因频率更高,差异有统计学意义(P0.05)。年龄65岁、内镜下见活动性出血、输血、再出血、CYP2C19*1/*2基因型、CYP2C19*1/*3基因型和CYP2C19*2等位基因是ANVUGIB患者死亡的危险因素(P0.05)。CYP2C19*1、*2和*3等位基因预测ANVUGIB患者死亡的曲线下面积分别为0.705、0.767、0.678。结论 CYP2C19基因多态性与ANVUGIB患者预后有关,CYP2C19*1/*2基因型、CYP2C19*1/*3基因型和CYP2C19*2等位基因是影响患者预后的危险因素。     

东莞地区汉族冠心病患者CYP2C19基因多态性分布特征分析

目的探讨东莞地区汉族冠心病患者CYP2C19基因多态性分布特征。方法选取该院2016年9月至2019年1月收治的800例汉族冠心病患者作为研究对象,采用等位基因特异PCR技术对患者CYP2C19基因多态性进行检测,了解CYP2C19基因多态性分布情况,并根据患者的年龄、性别进行分组分析。结果 CYP2C19基因型中*1/*1出现频率为38.50%,*1/*2频率为40.00%,*1/*3频率为6.38%,*2/*2频率为11.75%,*2/*3频率为3.00%,*3/*3频率为0.37%;其中快代谢型为38.50%、中代谢型为46.38%、慢代谢型为15.12%;等位基因分布中CYP2C19*1占61.69%,CYP2C19*2占33.25%,CYP2C19*3占5.06%。不同年龄、性别、患者CYP2C19各基因多态性分布虽存在一定的差异,但差异无统计学意义(P>0.05)。结论东莞地区汉族人群冠心病患者CYP2C19基因型中以快代谢型与中代谢型为主,基因分布以CYP2C19*1占比较高,不同性别、年龄患者CYP2C19基因型间差异不明显。     

Influence of genetics and non-genetic factors on acenocoumarol maintenance dose requirement in Moroccan patients

What is known and Objective: Coumarin derivatives such as acenocoumarol represent the therapy of choice for the long‐term treatment and prevention of thromboembolic diseases. Many genetic, clinical and demographic factors have been shown to influence the anticoagulant dosage. Our aim was to investigate the contribution of genetic and non‐genetic factors to variability in response to acenocoumarol in Moroccan patients. Methods: Our study included 114 adult Moroccan patients, receiving long‐term acenocoumarol therapy for various indications. Tests for VKORC1 ‐1639G>A promoter polymorphism (rs9923231), CYP2C9*2 rs1799853, CYP2C9*3 rs1057910, and CYP4F2 rs2108622 alleles were undertaken using Taq Man^® Pre‐Developed Assay Reagents for allelic discrimination. The statistical analysis was performed using the SAS V9 statistical package. Results and Discussion: Genotyping showed that the allele frequencies for the SNPs studied were no different to those found in Caucasians population. A significant association was observed between the weekly maintenance dose and the VKORC1 (P = 0·0027) and CYP2C9 variant genotypes (P = 0·0082). A final multivariate regression model that included the target International Normalized Ratio, VKORC1 and CYP2C9 genotypes explained 36·2% of the overall interindividual variability in acenocoumarol dose requirement. What is new and Conclusion: Our study shows large interindividual variability in acenocoumarol maintenance dose requirement in our population. VKORC1 and CYP2C9 variants significantly affected acenocoumarol dose, in‐line with results in other populations. For the Moroccan population, the SNPs that have the largest effect on acecoumarol dose are CYP2C9 rs1799853, CYP2C9 rs1057910 and VKORC1 rs9923231.     

Evaluation of the relationship between polymorphisms in CYP2C19 and the single‐dose pharmacokinetics of omeprazole in healthy Chinese volunteers: A multicenter study

The aim of this study was to evaluate the relationship between polymorphisms in CYP2C19 and the single‐dose pharmacokinetics (PKs) of omeprazole in healthy Chinese volunteers. A 20 mg single dose of omeprazole (Losec) enteric‐coated capsules or tablets was orally administered to 656 healthy subjects from eight subcenters. The polymorphic alleles of CYP2C19*2, *3, and *17 were determined by Sanger sequencing and Agena mass array. Plasma concentrations of omeprazole were determined by high‐performance liquid‐chromatography tandem mass spectrometry. PK parameters of area under the concentration versus time curve (AUC)_0‐t, AUC from zero to infinity (AUC_0‐∞), maximum plasma concentration (C_max), and terminal half‐life (t_1/2) were significantly influenced by CYP2C19 phenotype (all p < 0.001) and diplotype (all p < 0.001), and the same results were obtained in the subgroup analysis of the effects of diet and dosage form. The polymorphisms of CYP2C19*2(rs4244285; all PK parameters p < 0.001) and *3(rs4986893; p _Cmax = 0.020, and the p values of other PK parameters were less than 0.001) were significantly associated with the PKs of omeprazole. For CYP2C19*17 (rs12248560), only t_1/2 showed a significant correlation (p = 0.032), whereas other PK parameters did not. The present study demonstrated that the Pks of omeprazole is greatly influenced by CYP2C19.     

华南地区汉族人群心脑血管相关基因多态性分析

目的了解心脑血管相关基因细胞色素P450酶(CYP)2C19、CYP2C9、维生素K环氧化物还原酶复合体1(VKORC1)和乙醛脱氢酶2(ALDH2)基因的单核苷酸多态性频率在华南地区汉族人群分布的情况。方法采用芯片技术对252例心脑血管疾病患者的CYP2C19、ALDH2、CYP2C9以及VKORC1位点基因多态性进行检测,并统计男女间基因型频率的差异。结果 CYP2C19的野生型*1/*1占42.5%,突变型*1/*2占36.9%,*1/*3占7.9%,*2/*2占9.9%,*2/*3占1.6%,*3/*3占1.2%,在男女人群间各类基因型的发生率差异无统计学意义(P0.05);CYP2C9只检测到2种等位基因CYP2C9*1/*1和CYP2C9*1/*3,男性基因频率分别为98.6%和1.4%,女性基因频率分别为90.1%和9.9%,在男女人群间的分布差异有统计学意义(P0.05);VKORC1有3种基因型AA、AG和GG,频率分别为80.6%、18.7%和0.8%,在男女人群间的分布差异无统计学意义(P0.05);ALDH2有3种基因型GG、GL和LL型,频率分别为55.2%、37.3%和7.5%,在男女人群间的分布差异无统计学意义(P0.05)。结论 CYP2C9的多态性分布与性别有关,CYP2C19、VKORC1和ALDH2的多态性分布与性别无关。     

Pharmacogenetics of acenocoumarol in patients with extreme dose requirements

Summary. Background: There is currently intense debate as to whether pharmacogenetic algorithms for estimating the initial dose of coumarins provide a more accurate dose than the fixed‐dose approach. Recently, it has been suggested that the greatest benefit of pharmacogenetic algorithms is observed in patients with extreme dose requirements. Objectives: To identify clinical and genetic factors that better characterize patients who need extreme acenocoumarol doses for steady anticoagulation state. Patients/methods: We reviewed 9538 patients with a steady acenocoumarol dose from three Spanish hospitals, selecting 83 who took ≤ 5.00 mg week^?1 (percentile 5, p5) and 203 taking ≥ 30.00 mg week^?1 (p95). We also selected patients matched by gender and age taking 13.50–14.00 mg week^?1 (p50). We genotyped VKORC1 (rs9923231), CALU (rs1043550), GGCX (rs699664), CYP2C9 (rs1799853; rs1057910), CYP4F2 (rs2108622) and F7 (rs5742910) single‐nucleotide polymorphisms (SNPs). Results: Comparison between p5 and p95 revealed five parameters with significant differences: body surface area (BSA) (P = 0.006), age, VKORC1, CYP2C9 and CYP4F2 genotypes (all P < 0.001). First VKORC1, and second, CYP2C9 SNPs played a strong effect by determining extreme doses, particularly in p95. Only one out of 203 p95 had the VKORC1 A‐1639A genotype, but this subject was CYP2C9*1/*1. In contrast, nine out of 83 p5 carried the VKORC1 G‐1639G genotype, although six of them were CYP2C9*3 homozygotes and another two were heterozygotes. Surprisingly, CYP4F2 V433M SNP displayed prevalences that suggest that its influence might only be evident when patients are treated with high doses. Conclusion: Two clinical data, age and BSA, and three SNPs in the VKORC1, CYP2C9 and CYP4F2 genes strongly predict outlier patients treated with acenocoumarol.     

CYP2C9 and CYP2C19 genetic polymorphisms: frequencies in the south Indian population

The aim of the study was to establish the frequencies of CYP2C9*1, *2, *3 and CYP2C19*1, *2 and *3 in the south Indian population and to compare them with the inter-racial distribution of the CYP2C9 and CYP2C19 genetic polymorphisms. Genotyping analyses of CYP2C9 and CYP2C19 were conducted in unrelated, healthy volunteers from the three south Indian states of Andhra Pradesh, Karnataka and Kerala, by the polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP). The allele frequencies of the populations of these three states were then pooled with our previous genotyping data of Tamilians (also in south India), to arrive at the distribution of CYP2C9 and CYP2C19 alleles in the south Indian population. Frequencies of CYP2C9 and CYP2C19 alleles and genotypes among various populations were compared using the two-tailed Fisher's exact test. The frequencies of CYP2C9*1, *2 and *3 in the south Indian population were 0.88 (95% CI 0.85-0.91), 0.04 (95% CI 0.02-0.06) and 0.08 (95% CI 0.06-0.11), respectively. The frequencies of CYP2C9 genotypes *1/*1, *1/*2, *1/*3, *2/*2, *2/*3 and *3/*3 were 0.78 (95% CI 0.74-0.82), 0.05 (95% CI 0.03-0.07), 0.15 (95% CI 0.12-0.18), 0.01 (95% CI 0.0-0.02), 0.01 (95% CI 0.0-0.02) and 0.0, respectively. CYP2C19*1, *2 and *3 frequencies were 0.64 (95% CI 0.60-0.68), 0.35 (95% CI 0.31-0.39) and 0.01 (95% CI 0.0-0.03), respectively. As a result of a significant heterogeneity, the data on CYP2C19 genotype frequencies were not pooled. The frequency of CYP2C9*2 mutant alleles in south Indians was higher than in Chinese and Caucasians, while CYP2C9*3 was similar to Caucasians. CYP2C19*2 was higher than in other major populations reported so far. The relatively high CYP2C19 poor-metabolizer genotype frequency of 12.6% indicates that over 28 million south Indians are poor metabolizers of CYP2C19 substrates.     

高分辨熔解曲线分析CYP3A5基因多态性方法学的建立

目的通过建立一种较为简单灵敏的高分辨熔解曲线(HRM)方法,对CYP3A5单核苷酸多态性(SNP)位点进行测定,从而确定CYP3A5基因型,用于指导临床他克莫司的用药剂量。方法根据CYP3A5SNP位点基因组序列设计引物,利用HRM方法检测已知基因型标本,选取最佳引物,并通过同测序结果比对的方式确定该方法检测CYP3A5基因型的正确度、重复性以及灵敏度。结果利用该方法可以直接判断CYP3A5基因型,其检测结果与一代测序结果完全一致,重复性好、灵敏度高。结论 HRM方法可以用于CYP3A5基因型检测,为他克莫司的个体化用药提供可靠的评估依据。     

Association Between the CYP2C9 Genotype and Hypoglycemia Among Patients With Type 2 Diabetes Receiving Sulfonylurea Treatment: A Meta-analysis

PurposeTwo common variants, CYP2C9*2 (Arg144Cys, rs1799853) and CYP2C9*3 (Ile359Leu, rs1057910), are known to reduce the catalytic function of the CYP2C9 enzyme. Because impaired catalytic function is likely to affect sulfonylurea metabolism, it is predictable that CYP2C9 loss-of-function alleles may increase the risk of sulfonylurea-induced hypoglycemia. This systematic review and meta-analysis aimed to assess the association between CYP2C9 genotype and hypoglycemia among patients with type 2 diabetes mellitus (T2DM) receiving sulfonylurea.MethodsWe searched for studies on the association between CYP2C9 genotype and sulfonylurea-induced hypoglycemia among patients with T2DM, published through August 7, 2020, using PubMed, Web of Science, and EMBASE. Odds ratios (ORs) and 95% CIs were calculated.FindingsFive cohort studies and 2 case-control studies were included, and the total number of patients analyzed in this meta-analysis was 2769. The CYP2C9 variant alleles were associated with an increase in sulfonylurea-induced hypoglycemia compared with wild-type homozygote (OR = 1.24; 95% CI, 1.03–1.48). Compared with CYP2C9 wild-type homozygotes, CYP2C9*2 allele was associated with increased incidence of hypoglycemia (OR = 1.85; 95% CI, 1.18–2.89), whereas the CYP2C9*3 allele failed to show the statistical significance (OR = 1.67; 95% CI, 0.40–6.86; P = 0.48).ImplicationsOn the basis of these results, CYP2C9 genotyping may be useful for predicting the risk of hypoglycemia during sulfonylurea treatment for T2DM.