Division of Experimental and Translational Genetics, University of Missouri Kansas City School of Medicine, Kansas City, USA.
Department of Biomedical and Health Informatics, University of Missouri Kansas City School of Medicine, Kansas City, MO, USA.
Sci Rep. 2019 Feb 4;9(1):1396. doi: 10.1038/s41598-018-37940-6.
Acetaminophen (APAP) is a commonly used analgesic responsible for more than half of acute liver failure cases. Identification of previously unknown genetic risk factors would provide mechanistic insights and novel therapeutic targets for APAP-induced liver injury. This study used a genome-wide CRISPR-Cas9 screen to evaluate genes that are protective against, or cause susceptibility to, APAP-induced liver injury. HuH7 human hepatocellular carcinoma cells containing CRISPR-Cas9 gene knockouts were treated with 15 mM APAP for 30 minutes to 4 days. A gene expression profile was developed based on the 1) top screening hits, 2) overlap of expression data from APAP overdose studies, and 3) predicted affected biological pathways. We further demonstrated the implementation of intermediate time points for the identification of early and late response genes. This study illustrated the power of a genome-wide CRISPR-Cas9 screen to systematically identify novel genes involved in APAP-induced hepatotoxicity and to provide potential targets to develop novel therapeutic modalities.
对乙酰氨基酚（APAP）是一种常用的镇痛药，导致了超过一半的急性肝功能衰竭病例。确定以前未知的遗传风险因素将为 APAP 诱导的肝损伤提供机制见解和新的治疗靶点。本研究使用全基因组 CRISPR-Cas9 筛选来评估对 APAP 诱导的肝损伤具有保护作用或易感性的基因。含有 CRISPR-Cas9 基因敲除的 HuH7 人肝癌细胞用 15mM APAP 处理 30 分钟至 4 天。根据 1）顶级筛选命中，2）APAP 过量研究的表达数据重叠，和 3）预测受影响的生物途径，开发了一个基因表达谱。我们进一步证明了在识别早期和晚期反应基因时采用中间时间点的可行性。本研究说明了全基因组 CRISPR-Cas9 筛选在系统地鉴定参与 APAP 诱导的肝毒性的新基因方面的强大功能，并为开发新的治疗方式提供了潜在的靶点。
