Studies of DNA interstrand crosslink repair to improve crosslinking drug therapie

DNA链间交联修复改善交联药物治疗的研究

• 批准号: 8400213
• 负责人: JEAN GAUTIER
• 金额: $ 33.2万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-18 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8400213
• 关键词: 2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole; Address; Biochemistry; Biological Assay; Blood; Cell-Free System; Cells; Cisplatin; DNA; DNA Adducts; DNA Interstrand Crosslinking; DNA Repair; DNA biosynthesis; DNA lesion; DNA-Directed DNA Polymerase; Data; Dose-Limiting; Down-Regulation; Drug effect disorder; Drug resistance; Drug toxicity; ERCC1 gene; Engineering; Enzymes; Fluorescence; G1 Arrest; Genes; Genetic Recombination; Genetic Transcription; Goals; Homeostasis; Human; In Vitro; Incidence; Lesion; Light; Lymphoma; Maintenance; Malignant Neoplasms; Malignant neoplasm of ovary; Malignant neoplasm of testis; Mammalian Cell; Mitotic; Modeling; Molecular; Monitor; Mono-S; Non-Small-Cell Lung Carcinoma; Normal Cell; Patients; Pharmaceutical Preparations; Plasmids; Polymerase; Protocols documentation; Public Health; Reaction; Recurrent tumor; Reducing Agents; Research; Resistance; Role; S Phase; Site; Solutions; Stem cells; Structure; Surgical incisions; Testing; Toxic effect; Work; adduct; base; cancer cell; cell type; chemotherapeutic agent; chemotherapy; clinically relevant; crosslink; cytotoxic; drug efficacy; falls; improved; inhibitor/antagonist; innovation; insight; knock-down; melting; neoplastic cell; novel; nuclease; prevent; repair enzyme; repaired; research study; small molecule; standard of care; tumor

DESCRIPTION (provided by applicant): Drugs that generate DNA crosslinks are among the most effective cancer chemotherapeutic agents. These drugs fall in several classes of bi-functional molecules that generate DNA mono-adducts, intrastrand and interstrand crosslinks and are the standard of care for many malignancies. Interstrand crosslinks (ICLs) block many DNA transactions and are thought to be the cytotoxic lesions responsible for most crosslinking drug efficacy. Their efficacy however, varies among cell types or patients. There are two major challenges associated with these compounds: 1) dose-limiting toxicity, primarily in the blood, and 2) acquired resistance. The goal of this proposal is to characterize the mechanisms of ICL repair to better understand and ultimately improve the mode of action of crosslinking agent-based chemotherapy. ICLs are repaired during and outside of S-phase. Replication-independent ICL repair (RIR) is robust and critical for survival in treated mammalian cells. The proposed studies specifically address the mechanism of ICL repair in the absence of other DNA lesions and therefore focus on the most clinically relevant repair reaction triggered by crosslinking drugs. Specifically, we propose to identify the nucleases (Aim 1) and DNA polymerases (Aim 2) involved in RIR. Finally, we propose to evaluate the impact of replication-dependent and -independent ICL repair on crosslinking drugs efficacy. We hypothesize that targeting these factors could increase the sensitivity of tumor cells to crosslinking agents and reduce the incidence of resistance. Our approach will combine biochemistry in cell-free extracts with innovative ICL repair assays in normal and tumor cells. We anticipate that a better understanding of the molecular mechanisms of ICL repair will shed light on the contribution of ICL lesions to crosslinking drug toxicity and on the impact of DNA repair on resistance to crosslinking therapy. PUBLIC HEALTH RELEVANCE: The proposed research is highly relevant to public health. A better understanding of the enzymes responsible for DNA interstrand crosslink (ICL) repair is warranted to unravel the mechanism of action of drugs that generate ICLs work. It will also provide insight into why these drugs, which are routinely used in chemotherapeutic protocols, have variable efficacy among cell types or patients. It can help explain how resistance to these drugs arises. Finally, the proposed research will identify potential targets to sensitize cells and to prevent resistance to crosslinking drugs.

描述（由申请人提供）：产生DNA交联的药物是最有效的癌症化学治疗剂之一。这些药物属于产生DNA单吸引力，astrand和链间交联的几类双功能分子，并且是许多恶性肿瘤的护理标准。链间交叉链接（ICLS）阻止了许多DNA交易，被认为是负责大多数交联药物功效的细胞毒性病变。然而，它们的功效在细胞类型或患者之间有所不同。这些化合物有两个主要的挑战：1）限制剂量的毒性，主要是在血液中，而2）获得的耐药性。该建议的目的是表征ICL修复的机制，以更好地理解并最终改善基于交联剂的化学疗法的作用方式。在S期间和外部修复ICL。与复制无关的ICL修复（RIR）对于处理的哺乳动物细胞中的生存至关重要。拟议的研究专门针对没有其他DNA病变的ICL修复机制，因此着重于通过交联药物触发的最临床相关的修复反应。具体而言，我们建议鉴定参与RIR的核酸酶（AIM 1）和DNA聚合酶（AIM 2）。最后，我们建议评估复制依赖性和独立的ICL修复对交联药物功效的影响。我们假设靶向这些因素可能会增加肿瘤细胞对交联剂的敏感性并降低耐药性的发生。我们的方法将在无细胞提取物中的生物化学与正常细胞和肿瘤细胞中创新的ICL修复测定法相结合。我们预计，对ICL修复的分子机制有更好的了解将阐明ICL病变对交联药物毒性以及DNA修复对交联治疗的耐药性的影响。 公共卫生相关性：拟议的研究与公共卫生高度相关。有必要更好地理解负责DNA间交联（ICL）修复的酶，以揭示产生ICLS起作用的药物的作用机理。它还将提供有关这些药物在化学治疗方案中通常使用的这些药物在细胞类型或患者之间具有可变疗效的原因。它可以帮助解释如何产生对这些药物的抗性。最后，拟议的研究将确定潜在的靶标，以使细胞和 为了防止耐药性交联药。