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.

项目摘要在此提案中，我们旨在表征和确定持久细胞状态的机制。具体来说，我们发现，癌症持久细胞对新近的铁肉芽症具有特异性和潜在敏感发现的非凋亡细胞死亡程序涉及脂质氢过氧化物的有毒堆积。高毒性死亡可以通过gpx4的化学或遗传抑制来诱导，这是主要人清除脂质过氧化物的抗氧化剂。我们发现药物幼稚的父母癌细胞和未经转化的人上皮细胞对铁肉食不敏感，并提出了阐明分子基础，使细胞对铁铁作用的敏感性。这将通过细胞和分子方法包括高吞吐量遗传相互作用筛选。在AIM 1中，我们计划评估乳腺癌持久性细胞中其他观察结果的一般性癌症类型，包括黑色素瘤，卵巢癌和肺癌。我们将确定为什么持久者可能对铁铁作用敏感，而他们的亲本癌细胞是不敏感。这将通过测量抗氧化细胞代谢物来实现和辅因子（例如谷胱甘肽和铁），GPX4抑制后差异过氧化的脂质，ROS 信号通路和其他机械分析。这些实验将为 AIM 3中描述的遗传相互作用研究。在AIM 2中，我们还将确定GPX4在在癌细胞饰面，PDX和Syngeneic中，持久细胞存活和获得的耐药性免疫功能的乳腺癌和黑色素瘤的小鼠模型，持续性垂体诱导体内。预防体内持续细胞中诱导的肿瘤复发将是一个重要的促进针对GPX4的临床干预措施的进一步工作的结果。我们还将专注于确定持久状态背后的遗传相互作用的努力对铁铁作用的敏感性。在AIM 3中，我们将基于我们现有的努力，以开发一个平台进行极留基因相互作用图（EMAP）。这将有助于我们确定关系在我们的屏幕命中之间，希望鉴定基因和小分子之间的协同作用。此外，它可能有助于我们发现与可药基因的新协同相互作用。我们已经完成了试点EMAP分析以建立可行性，在这里我们建议扩展范围与整个基因组。癌症持久细胞的基础机制尚未彻底探索，该提案将确定基因和疾病相关性。我们的成就将产生第一个持久的细胞状态遗传相互作用图，并为识别多层用于翻译到诊所。