Plasticity of ether lipids promotes ferroptosis susceptibility and evasion

Ferroptosis, an iron-dependent, non-apoptotic cell death program, is involved in a variety of degenerative diseases and represents a targetable vulnerability in certain cancers. The ferroptosis-susceptible cell-state can either preexist in cells arising from certain lineages or be acquired during cell-state transitions. Precisely how ferroptosis susceptibility is dynamically regulated remains poorly understood. Using genome-wide CRISPR/Cas9 suppressor screens, we identify a key role for peroxisome–ether phospholipid genes in driving ferroptosis susceptibility and evasion in ovarian cancer. Overall design: Ovcar8 CRISPR library infected cells treated with escalating doses of RSL3 (Selleckchem) for 4 days each: 0.5 µM, 1 µM, and 2µM. 1×10^7 cells each were collected from the surviving population of RSL3-treated cells and an endpoint-matched untreated population for downstream sequencing analysis.

铁死亡（Ferroptosis）是一种铁依赖性非凋亡程序性细胞死亡程序，参与多种退行性疾病的发生发展，并可作为部分癌症的可靶向干预靶点。铁死亡易感细胞状态既可预先存在于特定谱系来源的细胞中，也可在细胞状态转换过程中获得。目前对于铁死亡易感性的动态调控机制仍不甚明晰。本研究通过全基因组CRISPR/Cas9抑制筛选，鉴定出过氧化物酶体醚磷脂基因在调控卵巢癌中铁死亡易感性与逃逸过程中的关键作用。总体实验设计：将感染CRISPR文库的Ovcar8细胞用梯度递增浓度的RSL3（Selleckchem）处理，每浓度处理4天，浓度依次为0.5 μM、1 μM与2 μM。分别收集存活的RSL3处理组与终点匹配的未处理组细胞各1×10^7个，用于后续测序分析。