Single Nucleotide Resolution Map of Formation and Repair of Bulky Adducts in the Human Genome

人类基因组中大体积加合物的形成和修复的单核苷酸解析图

• 批准号: 9186286
• 负责人: AZIZ SANCAR
• 金额: $ 46.84万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9186286
• 关键词: Address; Affect; Antineoplastic Agents; Area; Basic Science; Benzo(a)pyrene; Blood Cells; Carcinogens; Cell Cycle; Cell Cycle Stage; Cell Differentiation process; Cell Proliferation; Cells; Chemotherapy-Oncologic Procedure; Chromatin; Cisplatin; DNA; DNA Adduction; DNA Adducts; DNA Damage; DNA Modification Process; DNA Repair Enzymes; DNA Sequence; DNA lesion; Data; Development; Diagnosis; Disease susceptibility; Environmental Carcinogens; Event; Excision; Excision Repair; Genome; Genomic DNA; Genomics; Goals; Human; Human Genome; Individual Differences; Knowledge; Lead; Lesion; Link; Location; Malignant Neoplasms; Maps; Methodology; Methods; Mission; Mutagenesis; Nucleotide Excision Repair; Nucleotides; Outcome; Patients; Physiological; Platinum; Positioning Attribute; Predisposition; Prevention strategy; Public Health; Qualifying; Regulatory Pathway; Repair Enzymology; Research; Resolution; Risk; Site; Surgical incisions; System; Technology; Therapeutic Effect; UV induced; Ultraviolet Rays; United States National Institutes of Health; Work; Xenograft procedure; adduct; base; cancer prevention; cancer risk; cancer therapy; carcinogenesis; cell type; chemotherapeutic agent; chemotherapy; environmental carcinogenesis; functional status; genome-wide; genome-wide analysis; histone modification; human disease; improved; innovation; novel; novel strategies; programs; repair enzyme; repaired; response; stem cell differentiation; tool; whole genome

PROJECT SUMMARY/ABSTRACT Numerous environmental carcinogens and anti-cancer drugs form bulky base adducts in genomic DNA. However, the precise location of these lesions throughout the human genome is not known, and the factors that affect both damage formation and repair are difficult to study with currently available methodologies. Because the genomic location of damage and repair strongly influence the occurrence of pathological conditions, there is a need for new approaches for mapping damage and repair events across the entire genome. The long-term goal of our research program is to better understand how the nucleotide excision repair system targets the removal of bulky base adducts from DNA. The objective of this particular proposal is to further develop and apply novel tools for mapping carcinogen- and chemotherapy-induced DNA damage formation and repair throughout the genome and to identify the key factors that influence both the induction of DNA damage and the efficiency of damage removal by nucleotide excision repair. Our group’s background and expertise in the areas of DNA repair enzymology and genomics makes us uniquely qualified to address this issue. For this proposal, we will focus on identifying the precise locations of DNA base damage formed by the environmental carcinogens ultraviolet (UV) light and benzo[a]pyrene and by platinum-based cancer chemotherapies. We recently developed unique sequencing technologies that we have termed Damage-seq and XR-seq to provide high-resolution DNA sequence information on the formation and repair, respectively, of damage throughout the entire human genome. The rationale for the proposed research is that the ability to map damage and repair may reveal unexpected links between environmental carcinogens, mutagenesis, and human disease at specific genomic sites and suggest new strategies for diagnosing and treating human cancers. Our basic research on DNA adducts in cancer risk and prevention will be examined in the following four specific aims: 1) Method for Quantitative Mapping of DNA Damage Sites (Damage-seq) across the Whole Human Genome; 2) Method for Quantitative Mapping of Excision Repair (XR-seq) of the Whole Human Genome; 3) Genome-wide Analysis of Adduct Formation and Repair as a Function of Differentiation, Cell Cycle, and Chromatin States; and 4) Genome-wide Analysis of Adduct Formation and Repair in Human Biospecimens. The novel methods that we recently developed and will further optimize for mapping damage formation and repair will be used throughout this work. This proposal is innovative because it provides a new and unparalleled approach for characterizing DNA damage induced by environmental carcinogens and anti-cancer drugs. The proposed research is significant because it is expected to significantly expand our understanding of DNA damage formation and repair in the human genome at an unprecedented level of resolution. Ultimately, this knowledge has the potential to improve the prevention strategies for environmental carcinogenesis and to lead to the development of new tools for diagnosing and treating human cancer.

项目摘要/摘要 许多环境致癌物和抗癌药物在基因组DNA中形成庞大的基础加合物。 但是，这些病变在整个人类基因组中的确切位置尚不清楚，以及 影响损伤形成和维修都难以研究当前可用的方法。因为 损害和修复的基因组位置强烈影响病理状况的发生，存在 需要在整个基因组中绘制损害和修复事件的新方法。长期 我们的研究计划的目标是更好地了解核丁基惊喜维修系统如何针对 从DNA中去除笨重的基础加合物。该特定建议的目的是进一步发展和应用 绘制致癌物和化学疗法诱导的DNA损伤形成和修复的新工具 基因组并确定影响诱导DNA损伤和效率的关键因素 通过核苷酸惊喜修复去除损害。我们小组在DNA领域的背景和专业知识 维修酶学和基因组学使我们有唯一的资格解决这个问题。对于这个建议，我们将 专注于确定环境致癌物形成的DNA碱基损伤的精确位置 紫外线（UV）光和苯并[a] pyrene以及基于铂的癌症化学疗法。我们最近开发了 我们称之为损坏序言和XR-Seq的独特测序技术可提供高分辨率DNA 整个人类基因组中有关损害的形成和修复的序列信息。 拟议的研究的理由是，绘制损坏和维修的能力可能揭示出意外 特定基因组部位的环境致癌物，诱变和人类疾病之间的联系 为诊断和治疗人类癌症提出新的策略。我们关于DNA加合物的基础研究 癌症风险和预防将在以下四个特定目的中进行检查：1）定量的方法 在整个人类基因组中的DNA损伤位点（损伤seq）的映射； 2）定量的方法 整个人类基因组的切除修复（XR-SEQ）的映射； 3）添加基因组的分析 形成和修复是分化，细胞周期和染色质状态的函数； 4）全基因组 分析人类生物测量中的添加剂形成和修复。我们最近的新方法 在整个工作中，将使用开发并进一步优化用于映射损坏的形成和维修。 该建议具有创新性，因为它提供了一种表征DNA损害的新的无与伦比的方法 由环境致癌物和抗癌药物诱导。拟议的研究很重要，因为它 预计将显着扩展我们对人类基因组中DNA损伤形成和修复的理解 在空前的分辨率水平上。最终，这些知识有可能改善预防 环境致癌策略，并导致开发诊断和 治疗人类癌。