A development of oligonucleotide based DNA array for CYP2C9 genotyping for individual therapy in patients with valve replacement receiving warfarin therapy

摘要：本研究的目的是验证CYP2C9单核苷酸多态性（snp）在华法林患者中的作用，以避免栓塞和出血事件等副作用。研究对象为26例接受华法林治疗的瓣膜置换术患者（19例主动脉瓣，4例二尖瓣，3例主动脉瓣和二尖瓣）。使用新近开发的OligoARRAY系统确定CYP2C9基因snp的基因分型。国际标准化比率（PT-1NR）在自动分析仪系统上测量，ISI值为1.15。CYP2C9基因分型的准确性[CYP2C9^*2突变体；C416T (Argl44Cys)， CYP2C9^*3突变体；ALO61C (Ile359Leu)]经OligoARRAY直接测序验证。然而，在这26例患者中均未发现突变基因型。野生基因型患者的PT-FNR值AVR组为1.6±0.2 (n= 19)， MVR组为2±0.5 (n= 4)， AVR&MVR组为2.2±0.4 （n=3）。各组平均华法林剂量分别为2.8±0.8 mg、3±0.7 mg和2.7±1.3 mg。综上所述，OligoARRAY可方便可靠地进行CYP2C9基因分型，具有常规应用于临床检测华法林治疗前易感个体的潜力。第二章。日本和以色列人群中CYP2C9^*2 （Ari'l44Cys）和CYP2C9^*3 (lle3S9Leu) 2亚型的种族差异摘要：CYP2C9参与了华法林等许多临床重要底物药物的代谢。本研究利用新开发的寡核苷酸DNA阵列（OligoArray^R）对日本和以色列人群中CYP2C9^*2和CYP2C9^*3等位基因分布的种族差异进行了分析。研究对象包括147名日本人和388名以色列人（100名德系犹太人、99名也门犹太人、100名摩洛哥犹太人和89名利比亚犹太人）。采用OligoArray^R检测CYP2C9^*2 （Argl44Cys (416 A>C)，外显子3）和CYP2C9^*3 （Ile359Leu (1061 A>C)，外显子7）基因型。CYP2C9^*2基因型（CC/CT+TT）的频率在日本人中（1/0）（OR 0.02）显著低于利比亚犹太人（0.697/0.303）（OR 2.13; 95%CI 1.07-4.24），高于德系犹太人（0.83/0.17）、也门犹太人（0.899/0.101）、摩洛哥犹太人（0.81/0.19）。CYP2C9^*3基因型（AA/AC±CC）的频率在日本人（0.986/0.014）（OR 0.08）中较低，在利比亚犹太人（0.652/0.349）（OR 3.03; 95%CI 1.5 ~ 6.1）和摩洛哥犹太人（0.77/0.23）（OR 1.69; 95%CI 0.62 ~ 3.48）中较德系犹太人（0.83/0.17）、也门犹太人（0.899/0.101）中较高。因此，CYP2C9^*2 （Argl44Cys）和CYP2C9^*3 （Ile359Leu）突变体在日本人群中是罕见的，并且在四个犹太民族中表现出不同的频率。少

Title I. The clinical use of oli0onucleotide based DNA array for CYP2C9^*3 and CYP2C9^*2 L'enotvPin2 in patients receiving warfarin therapyAbstract : The aim of this investigation was to verify the contribution of CYP2C9 single nucleotide polymorphisms (SNPs) in patients receiving warfarin to avoid side effects such as embolic and hemorrhagic events. The subjects consisted of 26 patients with valve replacements (19 aortic valve, 4 mitral valve, 3 aortic and mitral valve), who were receiving warfarin therapy. Genotyping of CYP2C9 gene SNPs was determined using a recently developed OligoARRAY system. The international normalized ratio (PT-1NR) was measured on an auto-analyzer system using ISI value of 1.15. The accuracy of genotyping CYP2C9 [CYP2C9^*2 mutant; C416T (Argl44Cys), CYP2C9^*3 mutant; ALO61C (Ile359Leu)] by the OligoARRAY was verified by direct sequencing. However, no mutant genotypes were found in any of these 26 patients. The PT-FNR values for patients with the wild genotype … More was 1.6 ±0.2 in the AVR group (n='19), 2±0.5 in the MVR group (n4), and 2.2±0.4 in the AVR&MVR group (n=3). The average warfarin dose was 2.8 ± 0.8 mg, 3 ± 0.7 mg, and 2.7±1.3 mg, respectively for these group. In conclusion, the OligoARRAY was found to be convenient for the reliable genotyping of CYP2C9, and has the potential for routine use in the clinic for detecting susceptible individuals prior to warfarin treatment.Title II. Ethnic differences in CYP2C9^*2 (Ari'l44Cys) and CYP2C9^*3 (lle3S9Leu) 2enotvpes in Japanese and Israeli populationsAbstract : CYP2C9 contributes to the metabolism of a number of clinically important substrate drugs such as warfarin. In the present study, ethnic differences in the CYP2C9^*2 and CYP2C9^*3 allele distribution in Japanese and Israeli populations were evaluated a using newly developed oligonucleotide based DNA array (OligoArray^R). The population studied consisted of 147 Japanese and 388 Israeli donors (100 Ashkenazi Jews, 99 Yemenite Jews, 100 Moroccan Jews and 89 Libyan Jews). The CYP2C9^*2 [Argl44Cys (416 C>T), exon 3] and CYP2C9^*3 (Ile359Leu (1061 A>C), exon 7) genotypes were determined using an OligoArray^R. The frequencies of CYP2C9^*2 genotype (CC/CT+TT) was significantly lower in Japanese (1/0)(OR 0.02), and was higher in Libyan Jews (0.697/0.303) (OR 2.13; 95%CI 1.07-4.24) compared with those in Ashkenazi Jews (0.83/0.17), Yemenite Jews (0.899/0.101), Moroccan Jews (0.81/0.19). The frequencies of CYP2C9^*3 genotype (AA/AC±CC) was significantly lower in Japanese (0.986/0.014)(OR 0.08), and was higher in Libyan Jews (0.652/0.349) (OR 3.03; 95%CI 1.5-6.1) and Moroccan Jews (0.77/0.23)(OR 1.69; 95%CI 0.62-3.48) compaered with those in Ashkenazi Jews (0.83/0.17), Yemenite Jews (0.899/0.101). Thus, the CYP2C9^*2 (Argl44Cys) and CYP2C9^*3 (Ile359Leu) mutant were rare in the Japanese population, and showed different frequencies in the four Jewish ethnic groups examined. Less