Functional Networks for Persister Cell Sensitivities

持久细胞敏感性的功能网络

基本信息

批准号：
10226239
负责人：
MICHAEL T MCMANUS
金额：
$ 50.58万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10226239
关键词：
Antioxidants BRAF gene Bioavailable Breast Cancer Model Breast Melanoma Cancer Patient Cancer cell line Cell Death Cell Fraction Cell Line Cell Survival Cells Cessation of life Characteristics Chemicals Clinic Clinical Clustered Regularly Interspaced Short Palindromic Repeats Development Disabled Persons Disease Drug resistance ERBB2 gene Epithelial Cells Gene Expression Generations Genes Genetic Glutathione Goals Human Immunocompetent Implant Intervention Iron Knock-out Knockout Mice Leukemic Cell Lipid Peroxides Lipids Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of ovary Mammalian Cell Maps Masks Measurement Measures Mediating Melanoma Cell Methods Modeling Molecular NADP Neoplasm Metastasis Normal tissue morphology Pathway interactions Pharmaceutical Preparations Pharmacotherapy Proliferating Reactive Oxygen Species Recurrence Relapse Residual Cancers Residual state Role Signal Pathway Signal Transduction Testing Therapeutic Toxic effect Translations Tumor Volume Work Xenograft procedure acquired drug resistance cancer cell cancer recurrence cancer type cell type cofactor combinatorial dosage experimental study gene interaction genetic approach in vivo inhibitor/antagonist insight malignant breast neoplasm melanoma mouse model neoplastic cell novel novel strategies prevent programs small molecule synergism targeted treatment therapeutic target tool tumor whole genome

项目摘要

Project Summary In this proposal we aim to characterize and identify mechanisms for the persister cell state. Specifically, we have found that cancer persister cells are specifically and potently sensitive to ferroptosis, a newly discovered non-apoptotic cell death program which involves toxic buildup of lipid hydroperoxides. Ferroptotic death can be induced by chemical or genetic inhibition of GPX4, the primary human antioxidant which scavenges lipid peroxides. We have found that drug naïve parental cancer cells and nontransformed human epithelial cells are insensitive to ferroptosis and propose to elucidate the molecular basis for persister cell sensitivity to ferroptosis. This will be accomplished via cellular and molecular approaches including high throughput genetic interaction screens. In Aim 1, we plan to evaluate the generality of our observations in breast cancer persister cells in other cancer types including melanoma, ovarian and lung cancer. We will determine mechanisms for why persisters are potently and specifically sensitive to ferroptosis, while their parental cancer cells are insensitive. This will be accomplished by measurement of anti- and pro-oxidant cellular metabolites and cofactors (e.g. glutathione and iron), differentially peroxidated lipids upon GPX4 inhibition, ROS signaling pathways and other mechanistic analyses. These experiments will provide a framework for genetic interaction studies described in Aim 3. In Aim 2, we will also establish the role for GPX4 in persister cell survival and acquired drug resistance in cancer cell xenografts, PDXs and syngeneic immunocompetent mouse models of breast cancer and melanoma by inducing ferroptosis in persisters in vivo. Preventing tumor recurrence by inducing ferroptosis in persister cells in vivo will be a significant result to promote further work towards clinical intervention targeting GPX4. We will also focus on our efforts to identify the genetic interactions behind the persister state and its sensitivity to ferroptosis. In Aim 3 we will build on our existing efforts to develop a platform for conducting persister gene interaction maps (EMAPs). This is will help us identify the relationship between our screen hits, with the hope of identifying synergies among the genes and small molecules. In addition it may help us discover new synergistic interactions with druggable genes. We have already accomplished a pilot level EMAP analysis to establish feasibility, and here we propose to expand the scope to the whole genome. The mechanisms underlying cancer persister cells have not been thoroughly explored and this proposal will identify genes and disease relevance. Our accomplishments will yield the first persister cell state genetic interaction map and pave the way to identify polytherapies for translation to the clinic.

项目概要在本提案中，我们的目标是描述和识别持久细胞状态的机制。我们发现癌症持续细胞特别且强烈地易受铁死亡的影响，铁死亡是一种新的疾病发现了非凋亡细胞死亡程序，其中涉及脂质氢过氧化物的毒性积累。铁死亡可以通过化学或基因抑制 GPX4 来诱导，GPX4 是人类的主要细胞。我们发现药物幼稚的亲代癌细胞和抗氧化剂可以清除脂质过氧化物。未转化的人上皮细胞对铁死亡不敏感，并建议阐明持久细胞对铁死亡敏感性的分子基础这将通过细胞和细胞来实现。分子方法，包括高通量遗传相互作用筛选。在目标 1 中，我们计划评估我们在其他乳腺癌持续细胞中观察到的普遍性。癌症类型包括黑色素瘤、卵巢癌和肺癌，我们将确定原因的机制。持续细胞对铁死亡具有强效且特异的敏感性，而它们的亲本癌细胞则这将通过测量抗氧化和促氧化细胞代谢物来实现。和辅助因子（例如谷胱甘肽和铁）、GPX4 抑制后的差异过氧化脂质、ROS 这些实验将为信号通路和其他机制分析提供框架。目标 3 中描述的遗传相互作用研究。在目标 2 中，我们还将确定 GPX4 在癌细胞异种移植物、PDX 和同基因中的持续细胞存活和获得性耐药性通过诱导持久性铁死亡建立免疫活性小鼠乳腺癌和黑色素瘤模型通过在体内诱导存留细胞铁死亡来预防肿瘤复发将是一个重要的方法。结果将促进针对 GPX4 的临床干预的进一步工作。我们还将致力于确定持久状态及其背后的遗传相互作用。在目标 3 中，我们将在现有努力的基础上开发一个平台。进行持久基因相互作用图谱（EMAPS）这将帮助我们识别这种关系。在我们的屏幕点击之间，希望能够确定基因和小分子之间的协同作用。此外，它还可以帮助我们发现与可药物基因的新的协同相互作用。完成了试点级 EMAP 分析以确定可行性，在此我们建议扩大癌症持续细胞的机制尚未被阐明。彻底探索，该提案将确定基因和疾病的相关性。将产生第一个持久细胞状态遗传相互作用图，并为识别多疗法铺平道路翻译到诊所。