OMA1, DELE1 and HRI relay mitochondrial stress to the mammalian integrated stress response

The goal of this study is to understand how mamalian cells respond to the mitochondial stress. Using Quant-seq, we identifed an integrated stress response signature in the cells with compromized mitochondria. Furthermore, using an unbiased CRISPRi genetic screen, we uncovered a novel signaling pathway, in which OMA1, DELE1 and HRI are three major players that relay the mitochondrial stress to the integated stress response. To further study the cellular response in the absence of this pathway, we again used Quant-seq to uncover alternative pathways that might play a role during the mitochondrial stress response. Overall design: The Quant-seq experiments were performed using HEK293T cells with OMA1, DELE1, HRI KD via CRISPRi under both untreated or 1.25 ng/mL oligomycin (16hrs) treated conditions with 3 replicates for each sample. For WT type cells, 50 ug/mL Doxycyline treated and untreated cells were also included for Quant-seq experiments with 2 replicates.

本研究旨在解析哺乳动物细胞对线粒体应激的应答机制。我们借助Quant-seq技术，在线粒体功能受损的细胞中鉴定出整合应激反应特征。进一步通过无偏CRISPRi遗传筛选，我们发现了一条全新的信号通路：其中OMA1、DELE1与HRI是将线粒体应激信号传递至整合应激反应的三个核心调控因子。为深入探究该通路缺失状态下的细胞应答情况，我们再次采用Quant-seq技术，挖掘可能参与线粒体应激应答的替代通路。实验设计概述：本研究的Quant-seq实验以HEK293T细胞为实验材料，分别设置未经处理与1.25 ng/mL寡霉素（处理16小时）两种处理条件；其中通过CRISPRi介导OMA1、DELE1、HRI基因敲低的样本，每组均设置3次生物学重复。针对野生型细胞，本研究额外设置50 μg/mL多西环素处理与未处理的实验组，每组均设置2次生物学重复。