Development of genotyping method for functionally relevant variants of cytochromes P450 in cynomolgus macaques.
Development of genotyping method for functionally relevant variants of cytochromes P450 in cynomolgus macaques.
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食蟹猴细胞色素 P450 功能相关变体的基因分型方法的开发。
DOI：
10.1111/jvp.12443
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Yamazaki H.
中科院分区：
文献类型：
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作者：
Uno Y;Osada N;Sakurai S;Shimozawa N;Iwata T;Ikeo K;Yamazaki H.
CYP2C9、CYP2C76、CYP2D6、CYP3A4和CYP3A5是重要的药物代谢酶，在药物代谢研究中被广泛应用。CYP2C9（C . 334a >C）、CYP2C76（C . 449tg >A）、CYP2D6（C . 449tg >A）在肝脏中大量表达并代谢细胞色素P450底物。CYP3A4（IVS3 + 1G>del）和cyp3a5 (c。625A>T)实质上影响酶的代谢活性，因此是药物代谢研究中的重要变异。在这项研究中，开发了一种实时PCR方法对这些变异进行基因分型。通过对两种野生型、两种杂合型和两种纯合型dna进行基因分型，验证了方法的有效性，并对41只食蟹猕猴（来自柬埔寨、印度尼西亚、菲律宾和越南）的五种变异以及另一种重要变异cyp2c19 （c.308C>T）进行基因分型。cyp2c9和cyp2c19变体仅在柬埔寨和越南动物中发现，而yp2c76和cyp2d6变体仅在印度尼西亚和菲律宾动物中发现。在分析的所有动物中均未发现cyp3a4和cyp3a5变异。毛里求斯动物使用下一代测序数据进行基因分型进行比较，具有yp2c19和cyp2d6变体，但没有其他变体。这些结果表明，这些重要变异在动物群体中的流行程度存在差异。因此，所开发的基因分型工具可用于食蟹猴的药物代谢研究。
In cynomolgus macaques (Macaca fascicularis), widely used in drug metabolism studies, CYP2C9, CYP2C76, CYP2D6, CYP3A4, and CYP3A5, important drug‐metabolizing enzymes, are abundantly expressed in liver and metabolize cytochrome P450 substrates.CYP2C9(c.334A>C),CYP2C76(c.449TG>A),CYP2D6(c.891A>G),CYP3A4(IVS3 + 1G>del), andCYP3A5(c.625A>T) substantially influence metabolic activity of enzymes, and thus are important variants in drug metabolism studies. In this study, a real‐time PCR method was developed for genotyping these variants. The validity of the methods was verified by genotyping two wild type, two heterozygous, and two homozygous DNAs and was used to genotype 41 cynomolgus macaques (from Cambodia, Indonesia, the Philippines, or Vietnam) for the five variants, along with another important variantCYP2C19(c.308C>T). TheCYP2C9andCYP2C19variants were found only in Cambodian and Vietnamese animals, while theCYP2C76andCYP2D6variants were found only in Indonesian and Philippine animals. TheCYP3A4andCYP3A5variants were not found in any of the animals analyzed. Mauritian animals, genotyped using next‐generation sequencing data for comparison, possessed theCYP2C19andCYP2D6variants, but not the other variants. These results indicated differences in prevalence of these important variants among animal groups. Therefore, the genotyping tool developed is useful for drug metabolism studies using cynomolgus macaques.