CRL3KEAP1 complex licenses genotoxic stress-induced tumor cell death

CRL3KEAP1复合物许可基因毒性应激诱导的肿瘤细胞死亡

基本信息

批准号：
10378636
负责人：
Yi-Nan Gong
金额：
$ 19.72万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10378636
关键词：
Antioxidants Apoptosis Apoptosis Inhibitor Apoptotic BIRC4 gene CASP8 gene CRISPR screen Cell Death Cell Line Cell Survival Cells Clustered Regularly Interspaced Short Palindromic Repeats Colorectal Neoplasms Complex DNA Damage Data Genotoxic Stress HT29 Cells Human Knock-out Knowledge Licensing Link Malignant Neoplasms Mediating Mediator of activation protein Membrane Modeling Molecular Mutate Mutation Necrosis Neoplasm Metastasis Pathway interactions Phosphorylation Phosphotransferases Positioning Attribute Predisposition Protein Kinase Proteins RIPK1 gene RIPK3 gene Radiation therapy Reagent Reporting Resistance Signal Pathway Signaling Molecule System TNF gene TP53 gene Testing UV induced base cancer cell cancer therapy cell killing chemotherapy genome-wide genotoxicity insight neoplastic cell novel overexpression refractory cancer response screening therapy resistant tumor tumor progression ubiquitin-protein ligase

项目摘要

PROJECT SUMMARY/ABSTRACT This application is responsive to PA-17-449: The Interplay of Cell Death Pathways in Cancer Cell Survival and Resistance to Therapy (R21). In general, genotoxic stress triggers apoptosis if tumor cells express functional p53. Genotoxic stress can also induce p53 independent apoptosis due to the auto-depletion of XIAP and cIAP1/2, which leads to the formation of “ripoptosome” complex. Ripoptosome can stimulate caspase-8-mediated apoptosis. Mutated p53 and caspase-8 have been frequently detected in human cancers. Caspase-8 mutation or inactivation often leads to resistance to apoptosis. However, in a few cases, it renders cells highly susceptible to another form of programmed cell death, termed “necroptosis.” Kinase RIPK3 and its substrate MLKL are the critical players for necroptosis. Although several studies have indicated that DNA damage can induce necroptosis (or undefined necrosis), the molecular mechanism is largely unknown. Herein, we have used an unbiased genome-wide CRISPR knockout approach to identify new players in DNA damage-induced necroptosis. We identified Cullin3 (Cul3)-RING E3 ubiquitin Ligase (CRL3KEAP1) complex as a promising candidate that mediated DNA damage-induced necroptosis. Our working model is that CRL3KEAP1 complex needs to degrade an unidentified substrate(s) to let the progression of DNA damage-induced necroptosis, thus licensing genotoxicity-induced cell death. We will make efforts to test this hypothesis in this proposal. We will determine what the CRL3KEAP1 substrate(s) is(are) and how it(they) can halt cell death. The proposed studies will provide new mechanistic insights on DNA damage agents induced cell death. Our ongoing efforts have the potential to change how we trigger cell death in tumors, especially in those with p53, caspase-8, and/or KEAP1 mutations.

项目概要/摘要此应用程序响应 PA-17-449：癌细胞中细胞死亡途径的相互作用生存和对治疗的抵抗（R21）一般来说，如果肿瘤，基因毒性应激会引发细胞凋亡。细胞表达功能性 p53，基因毒性应激也可诱导 p53 不依赖的细胞凋亡。 XIAP 和 cIAP1/2 的自动耗尽，导致“核糖体”复合物的形成。 Ripoptosome 可以刺激 caspase-8 介导的细胞凋亡。突变的 p53 和 caspase-8 有。 Caspase-8 突变或失活经常导致人类癌症中被发现。然而，在少数情况下，它使细胞对另一种细胞高度敏感。一种程序性细胞死亡，称为“坏死性凋亡”。激酶 RIPK3 及其底物 MLKL 是尽管一些研究表明 DNA 损伤可以导致坏死性凋亡。诱导坏死性凋亡（或未定义的坏死），其分子机制很大程度上未知。我们使用了无偏见的全基因组 CRISPR 敲除方法来识别新的参与者我们鉴定了 Cullin3 (Cul3)-RING E3 泛素连接酶。 (CRL3KEAP1) 复合物作为介导 DNA 损伤诱导的坏死性凋亡的有希望的候选者。我们的工作模型是 CRL3KEAP1 复合体需要将未识别的底物降解为让 DNA 损伤诱导的坏死性凋亡进展，从而许可基因毒性诱导的细胞我们将努力在这个提案中检验这个假设，我们将确定什么。 CRL3KEAP1 底物是什么以及它如何阻止细胞死亡？为 DNA 损伤剂诱导细胞死亡提供新的机制见解。有潜力改变我们触发肿瘤细胞死亡的方式，特别是那些带有p53的肿瘤， caspase-8 和/或 KEAP1 突变。