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: 9976511
• 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 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9976511
• 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; DNA mapping; 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; Pathogenicity; Pathologic; Patients; Physiological; Platinum; Positioning Attribute; Predisposition; Prevention strategy; Public Health; Regulatory Pathway; Repair Enzymology; Research; Resolution; Risk; Site; Structure; Surgical incisions; System; Technology; Therapeutic Effect; UV induced; Ultraviolet Rays; United States National Institutes of Health; Work; Xenograft procedure; adduct; anti-cancer; base; cancer prevention; cancer risk; cancer therapy; carcinogenesis; carcinogenicity; 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.

项目总结/摘要 许多环境致癌物和抗癌药物在基因组DNA中形成庞大的碱基加合物。 然而，这些病变在整个人类基因组中的精确位置尚不清楚， 影响损伤形成和修复的因素难以用现有方法研究。因为 损伤和修复的基因组定位强烈影响病理状况的发生， 需要新的方法来绘制整个基因组的损伤和修复事件。长期 我们的研究计划的目标是更好地了解核苷酸切除修复系统如何靶向 从DNA中去除大体积的碱基加合物。这项建议的目的是进一步发展和应用 用于绘制致癌物和化疗诱导的DNA损伤形成和修复的新工具 基因组，并确定影响DNA损伤诱导和效率的关键因素 通过核苷酸切除修复来去除损伤。我们团队在DNA领域的背景和专业知识 修复酶学和基因组学使我们有独特的资格来解决这个问题。对于这一建议，我们将 重点是确定环境致癌物形成的DNA碱基损伤的精确位置 紫外线（UV）和苯并[a]芘以及基于铂的癌症化疗。我们最近开发 我们称之为Damage-seq和XR-seq的独特测序技术， 在整个人类基因组中分别形成和修复损伤的序列信息。 拟议研究的基本原理是，绘制损伤和修复的能力可能会揭示意想不到的 环境致癌物、诱变和人类疾病在特定基因组位点之间的联系， 提出了诊断和治疗人类癌症的新策略。我们对DNA加合物的基础研究 癌症风险和预防将在以下四个具体目标进行审查：1）定量方法 整个人类基因组中DNA损伤位点的定位（Damage-seq）; 2）定量分析方法 全人类基因组的切除修复定位（XR-seq）; 3）加合物的全基因组分析 形成和修复是分化、细胞周期和染色质状态的函数;和4）全基因组 人体生物标本中加合物形成和修复的分析。我们最近使用的新方法 开发并将进一步优化映射损伤形成和修复将在整个工作中使用。 这一建议是创新的，因为它提供了一个新的和无与伦比的方法来表征DNA损伤 由环境致癌物和抗癌药物引起。这项研究意义重大，因为它 有望大大扩展我们对人类基因组中DNA损伤形成和修复的理解 以前所未有的分辨率。最终，这些知识有可能改善预防 环境致癌作用的战略，并导致新的诊断工具的发展， 治疗人类癌症