Massively parallel characterization of CYP2C9 variant enzyme activity and abundance
Massively parallel characterization of CYP2C9 variant enzyme activity and abundance
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DOI：
10.1016/j.ajhg.2021.07.001
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发表时间：
2021-09-02
影响因子：
9.8
通讯作者：
Dunham, Maitreya J.
中科院分区：
文献类型：
--
作者：
Amorosi, Clara J.;Chiasson, Melissa A.;Dunham, Maitreya J.
CYP 2C 9编码细胞色素P450酶，负责代谢高达15%的小分子药物，CYP 2C 9变体可以改变这些治疗药物的安全性和有效性。特别是，每年有超过1500万人服用抗凝剂华法林，CYP 2C 9的多态性可影响个体药物反应并导致出血风险增加。我们开发了clickseq，一种基于酵母的活性检测方法，用于测试数千种变体。使用click-seq，我们测量了酵母中6,142个错义变体的活性。我们还通过大规模平行测序（VAMP-seq）使用变体丰度测量了人类细胞系中6，370个错义变体的稳态细胞丰度。这些数据显示，几乎三分之二的CYP 2C 9变体显示活性降低，蛋白质丰度占CYP 2C 9功能变异的一半。我们还测量了319个以前未注释的人类变异的活性评分，其中许多可能具有临床相关性。
CYP2C9 encodes a cytochrome P450 enzyme responsible for metabolizing up to 15% of small molecule drugs, and CYP2C9 variants can alter the safety and efficacy of these therapeutics. In particular, the anti-coagulant warfarin is prescribed to over 15 million people annually and polymorphisms in CYP2C9 can affect individual drug response and lead to an increased risk of hemorrhage. We developed clickseq, a pooled yeast-based activity assay, to test thousands of variants. Using click-seq, we measured the activity of 6,142 missense variants in yeast. We also measured the steady-state cellular abundance of 6,370 missense variants in a human cell line by using variant abundance by massively parallel sequencing (VAMP-seq). These data revealed that almost two-thirds of CYP2C9 variants showed decreased activity and that protein abundance accounted for half of the variation in CYP2C9 function. We also measured activity scores for 319 previously unannotated human variants, many of which may have clinical relevance.