The roles of anti-apoptotic proteins BCL-XL and MCL1 in mediating survival of high-grade serous ovarian cancer following drug-induced DNA damage

项目摘要/摘要 高级别浆液性卵巢癌(HGSOC)通常诊断为晚期，预后较差。 虽然许多患者最初对铂和紫杉醇化疗有反应，但大多数患者 对化疗有抗药性的复发性疾病。化疗耐药是一种未得到满足的治疗需求 HGSOC。为了应对这一挑战，斯托弗博士进行了系统的功能基因组筛查，以确定 铂类和紫杉醇耐药的中介物。保护细胞免受凋亡性死亡的抗凋亡蛋白 与化疗耐药密切相关。相反，抑制抗细胞凋亡的靶向药物 BCL-XL和MCL1蛋白使HGSOC细胞对化疗敏感。在这项提案中，斯托弗博士将测试 BCL-XL和MCL1是化疗诱导DNA后HGSOC存活的关键介质假说 损坏。目标1将确定bclxl和mcl1抑制剂联合应用时是否会增加细胞凋亡 DNA损伤化疗在HGSOC细胞株和患者细胞中的应用及DNA损伤效应探讨 BCL-XL和MCL1与促凋亡蛋白相互作用的药物。目标2将评估DNA是否 HGSOC细胞的损伤修复能力影响其生存对bclxl和mcl1的依赖。一个 化学筛选将旨在识别DNA损伤剂和其他与bclxl和bclxl有协同作用的药物 MCL1抑制剂可杀死HGSOC细胞。目的3将评估bcl-xl和mcl1在肺癌生存中的作用。 耐化疗的HGSOC。BCL-XL和MCL1抑制剂与化疗的组合将进行测试 在体外经验性铂耐药的HGSOC细胞系和HGSOC细胞系异种移植瘤和患者来源的HGSOC细胞系中 体内异种移植。最后，我们将使用基因组和蛋白质组方法来分析患者样本 未治疗和化疗耐药的HGSOC对bclxl和mcl1及相关抗细胞凋亡的影响 蛋白质，以及其他化疗耐药途径。总而言之，这项研究将评估需求 BCL-XL和MCL1在HGSOC化疗耐药中的作用，并开始阐明两者之间的串扰 耐药中的细胞凋亡和DNA损伤修复途径。BCL-XL或MCL1抑制剂与 DNA损伤剂可能是治疗HGSOC的有效方法。斯托弗博士的长期目标是为 通过专注于卵巢癌的独立研究为卵巢癌患者提供新的治疗选择 生物学、基因组学和治疗学。这一目标的实现将得到杰出的机构的推动 Dana-Farber癌症研究所的环境和包括Matthew Meyerson博士在内的有成就的导师， 医学博士和Anthony Letai博士。与布罗德研究所的合作将做出非凡的贡献 临床基因组学和化学筛查的资源，以及一个专家咨询委员会将提供指导 关于实验设计和职业发展。研究培训计划，包括课程和指导 在关键的生物学主题和专业发展方面，将支持斯托弗博士独立研究的目标 从事以临床为重点的实验室研究，以改善卵巢癌的预后。

PROJECT SUMMARY/ABSTRACT High-grade serous ovarian cancer (HGSOC) is often diagnosed at an advanced stage and has a poor prognosis. Although many patients initially respond to platinum and paclitaxel chemotherapy, most patients develop recurrent disease that is resistant to chemotherapy. Chemotherapy resistance is an unmet therapeutic need in HGSOC. To address this challenge, Dr. Stover performed systematic functional genomic screens to identify mediators of platinum and paclitaxel resistance. Anti-apoptotic proteins that protect cells from apoptotic cell death were strongly associated with chemotherapy resistance. Conversely, targeted drugs that inhibit anti-apoptotic proteins BCL-XL and MCL1 sensitized HGSOC cells to chemotherapy. In this proposal, Dr. Stover will test the hypothesis that BCL-XL and MCL1 are key mediators of HGSOC survival following chemotherapy-induced DNA damage. Aim 1 will determine whether BCL-XL and MCL1 inhibitors increase apoptosis when combined with DNA-damaging chemotherapy in HGSOC cell lines and patient cells, and explore effects of DNA-damaging agents on BCL-XL and MCL1 interactions with pro-apoptotic proteins. Aim 2 will assess whether the DNA damage repair capacity of HGSOC cells affects their dependency upon BCL-XL and MCL1 for survival. A chemical screen will aim to identify DNA-damaging agents and other drugs that synergize with BCL-XL and MCL1 inhibitors to kill HGSOC cells. Aim 3 will evaluate the role of BCL-XL and MCL1 in the survival of chemotherapy-resistant HGSOC. Combinations of BCL-XL and MCL1 inhibitors and chemotherapy will be tested in HGSOC cell lines with empiric platinum resistance in vitro, and HGSOC cell-line xenografts and patient-derived xenografts in vivo. Finally, we will use genomic and proteomic approaches to analyze patient samples of untreated and chemotherapy-resistant HGSOC for changes in BCL-XL and MCL1 and related anti-apoptotic proteins, as well as other chemotherapy resistance pathways. In summary, this study will assess the requirement for BCL-XL and MCL1 in HGSOC chemotherapy resistance, and begin to elucidate the crosstalk between apoptosis and DNA damage repair pathways in resistance. Combinations of BCL-XL or MCL1 inhibitors with DNA-damaging agents may be effective therapies for HGSOC. Dr. Stover's long-term goal is to contribute to new treatment options for patients with ovarian cancer through independent research focusing on ovarian cancer biology, genomics, and therapeutics. The pursuit of this goal will be facilitated by the outstanding institutional environment of the Dana-Farber Cancer Institute and accomplished mentors, including Drs. Matthew Meyerson, MD PhD and Anthony Letai, MD PhD. Collaborations with the Broad Institute will contribute exceptional resources for clinical genomics and chemical screening, and an expert advisory committee will provide guidance on experimental design and career advancement. A research training program, including courses and mentoring in key biological topics and professional development, will support Dr. Stover's goal of an independent research career pursuing laboratory research with a clinical focus to improve outcomes in ovarian cancer.