Exploitation of Intrinsic DNA Repair Defects with DNA Damaging Agents in Cancer

利用 DNA 损伤剂治疗癌症中的内在 DNA 修复缺陷

• 批准号: 10441362
• 负责人: Kingson Lin
• 金额: $ 3.16万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-16 至 2023-07-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10441362
• 关键词: Adjuvant Chemotherapy; Age; Alkylating Agents; Alkylation; Biological Assay; Biology; Biotechnology; CRISPR/Cas technology; Cancer Biology; Cancer Burden; Cell Death; Cell Line; Cell model; Cell-Free System; Cells; Chemicals; Chemistry; Chemotherapy-Oncologic Procedure; Clinic; Clinical; Collection; Colorectal Cancer; DNA; DNA Alkylation; DNA Damage; DNA Repair; DNA Repair Pathway; Data; Defect; Development; Dioxygenases; Disease; Engineering; Enzymes; Epigenetic Process; Exhibits; Financial Hardship; Glioblastoma; Glioma; Goals; Hematologic Neoplasms; In Vitro; Isocitrate Dehydrogenase; Isocitrates; Knowledge; Laboratories; Lymphoma; Malignant Neoplasms; Mechlorethamine; Mentorship; Methodology; Methyltransferase; Methyltransferase Gene; Modeling; Modification; Mustard Gas; Mutation; Nature; Organic Chemistry; Outcome; Pathway interactions; Patients; Pattern; Physicians; Population; Protein Family; Radiation therapy; Recording of previous events; Regimen; Research Activity; Resistance; Resources; Scientist; Survival Rate; Techniques; Testing; Therapeutic; Time; Training; Treatment Protocols; Treatment outcome; Tumor Suppressor Genes; Work; adduct; alpha ketoglutarate; base; cancer cell; cancer risk; cancer therapy; cancer type; chemotherapy; cohesion; cytotoxic; experience; experimental study; genome analysis; improved; in vivo; inhibitor; insight; interdisciplinary approach; leukemia; lifetime risk; liquid chromatography mass spectroscopy; mutant; neoplastic cell; novel; plasmid DNA; rational design; repair enzyme; repaired; response; screening; standard of care; targeted treatment; tool; tumor; tumor DNA

Project Summary: Exploitation of Intrinsic DNA Repair Defects with DNA Damaging Agents in Cancer The lifetime risk of cancer in the U.S. is 1 in 3 and the financial burden of cancer exceeds 100 billion dollars annually. These figures are predicted to continue increasing as populations continue to age. Alkylating agents were the first chemotherapies used to treat cancer with the application of nitrogen mustard gas to treat lymphoma in the 1940s at Yale. Over the last 80 years, many different classes of alkylating agents have been developed and they remain an integral component of cancer treatment today. Despite their long history and prevalent clinical usage, knowledge of how alkylating agents damage DNA is still poorly understood due to the reactive nature of these species. Additionally, cancers often develop exploitable therapeutic vulnerabilities in DNA repair pathways that enable them to accumulate more mutations and become more aggressive. This incomplete understanding of alkylators results in the empirical selection of alkylating agents in cancer treatment regimens rather than selection based on mechanism and underlying cancer biology. MGMT and ALKBH2/3 are two key direct DNA alkyl damage reversal enzymes responsible for repairing a variety of alkyl adducts. These enzymes are also commonly deficient in isocitrate dehydrogenase1/2 (IDH1/2) mutant cancers such as gliomas, colorectal cancers, and hematological malignancies. Understanding how alkylators damage DNA in the absence of any repair enzymes and how repair enzymes contribute to alkylator resistance is crucial for the therapeutic advancement of alkylating chemotherapies including a better understanding of alkylating agents, the development of novel alkylators, and personalized alkylator selection based on patient tumor DNA repair status. I will test the hypothesis that targeting cancer cells deficient in either MGMT and/or ALKBH2/3 with the appropriate alkylator will result in enhanced sensitivity because deficiency in the corresponding DNA repair pathway will lead to unrepairable damage. I plan to test this hypothesis through two aims. My first aim is to develop a LCMS-based assay to profile the spectrum of alkylation damage in cell free plasmid DNA and CRISPR/Cas9 generated glioma models. This aim will answer the question of how alkylators damage DNA by identifying both the species and quantities of DNA alkylation adducts that form when cell free plasmid DNA and various glioma model cell lines are treated with a panel of clinically used alkylators. My second aim is to conduct a high-throughput differential screen for alkylator sensitivity based on DNA repair pathway status. This aim will elucidate the relationship between known DNA repair pathways and alkylator sensitivity. Ultimately, this work could contribute to the therapeutic advancement of alkylating chemotherapies including the development of novel alkylators and personalized alkylator selection based on patient tumor DNA repair status.

项目概要：利用DNA损伤剂对癌症中的固有DNA修复缺陷进行研究 在美国，癌症的终生风险是三分之一，癌症的经济负担超过1000亿美元。 美元每年。预计随着人口继续老龄化，这些数字将继续增加。烷基化 药物是第一种用于治疗癌症的化学疗法，其中应用氮芥子气治疗癌症。 1940年代在耶鲁大学发现的淋巴瘤在过去的80年中，已经研究了许多不同种类的烷基化剂。 它们已经被开发出来，并且今天仍然是癌症治疗的组成部分。尽管历史悠久， 尽管普遍的临床使用，但由于烷化剂对DNA的损伤， 这些物种的反应性。此外，癌症通常在治疗过程中发展出可利用的治疗弱点。 DNA修复途径，使它们能够积累更多的突变，变得更具侵略性。这 对烷化剂的不完全理解导致在癌症治疗中经验性地选择烷化剂 治疗方案，而不是基于机制和潜在的癌症生物学的选择。 MGMT和ALKBH 2/3是两个关键的直接DNA烷基损伤逆转酶，负责修复 各种烷基加合物。这些酶通常也缺乏异柠檬酸脱氢酶1/2（IDH 1/2） 突变型癌症如神经胶质瘤、结肠直肠癌和血液恶性肿瘤。了解如何 在没有任何修复酶的情况下，烷化剂损伤DNA，以及修复酶如何促进烷化剂 耐药性对于烷基化化疗的治疗进展是至关重要的， 对烷基化剂的理解，新型烷基化剂的开发，以及个性化的烷基化剂选择 基于患者肿瘤DNA修复状态。我将验证一个假设，即靶向缺乏 MGMT和/或ALKBH 2/3与适当的烷基化剂将导致敏感性增强，因为 相应的DNA修复途径将导致不可修复的损伤。 我打算通过两个目标来检验这个假设。我的第一个目标是开发一种基于LCMS的检测方法， 分析无细胞质粒DNA和CRISPR/Cas9产生的胶质瘤中烷基化损伤的谱 模型这一目标将通过识别物种和DNA来回答烷化剂如何破坏DNA的问题。 当无细胞质粒DNA和各种胶质瘤模型细胞系时形成的DNA烷基化加合物的量 用一组临床上使用的烷化剂处理。我的第二个目标是进行高通量的 基于DNA修复途径状态的烷化剂敏感性的差异筛选。这一目标将阐明 已知的DNA修复途径和烷化剂敏感性之间的关系。最终，这项工作将有助于 涉及烷基化化学疗法的治疗进展，包括新型烷基化剂的开发， 基于患者肿瘤DNA修复状态的个性化烷化剂选择。