无意义基因变异对OATP1B1表达和输送活动的 in vitro 影响
Wilma Kiander1, Hatam Rashidpour2, Mikko Gynther3
1Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland; Department of Clinical Pharmacology, Faculty of Medicine, University of Helsinki, Helsinki, Finland; Individualized Drug Therapy Research Program, Faculty of Medicine, University of Helsinki, Helsinki, Finland; Department of Clinical Pharmacology, HUS Diagnostic Centre, Helsinki University Hospital, Helsinki, Finland.
研究SLCO1B1基因变异,这项研究揭示了大多数无意义的变异显著减少有机离子运输聚1B1 (OATP1B1) 功能和膜表达,可能会增加药物的血度.
科学领域:
- 药物基因组学 药物基因组学
- 分子生物学分子生物学
- 药物代谢药物代谢
背景情况:
- 有机离子运输聚1B1 (OATP1B1) 对于肝脏药物和化合物清除至关重要.
- 在SLCO1B1基因的遗传变异可以改变OATP1B1载体活性和血药物水平.
- 无意义的变异和功能丧失突变与像罗托氏综合征这样的疾病有关.
研究的目的:
- 调查选择的SLCO1B1无意义变异对OATP1B1传输活动和膜表达的功能影响.
- 为了确定特定的变体位置是否与保留或丢失的传送器功能相关.
主要方法:
- 使用编码变体或参考SLCO1B1序列的baculovirus构造来设计HEK293细胞.
- 评估了OATP1B1的运输活动,使用2',7'-二二素和罗斯瓦斯塔丁的吸收试验.
- 使用定量定向绝对蛋白质组学来测量OATP1B1蛋白质的丰度.
主要成果:
- 没有意义的变体c.1968dup和c.1976C>G保留了30-40%的OATP1B1运输活动.
- 其他研究的变体,位于较早或最终的跨膜螺旋体内,导致OATP1B1功能完全丧失.
- 所有测试的变体都导致细胞膜OATP1B1表达的减少.
结论:
- 大多数SLCO1B1无意义变体导致OATP1B1活动的显著损失.
- 靠近C端的变体可能保留部分OATP1B1传输功能.
- 携带这些变异的个体可能会出现OATP1B1基质药物的血度升高.
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