Ultrasensitive measure of human mutagenesis: Connecting the exposome to disease

人类诱变的超灵敏测量：将暴露组与疾病联系起来

• 批准号: 9176699
• 负责人: Jason H Bielas
• 金额: $ 39.6万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9176699
• 关键词: Acute; Address; Age; Aging; Architecture; Biological Assay; Biological Markers; Blood; Cell Proliferation; Cells; Chemopreventive Agent; China; Chronic; Coal; Complex; Coupled; Cross-Sectional Studies; DNA; DNA Adducts; DNA Repair; Data; Detection; Development; Disease; Early Diagnosis; Early identification; Environmental Exposure; Environmental Risk Factor; Etiology; Event; Excision; Exposure to; Foundations; Frequencies; Genes; Genetic Predisposition to Disease; Genetic Transcription; Genetic Variation; Genome; Genomic Segment; Genomics; Goals; Heating; Human; Human Genetics; Human Genome; Incidence; Individual; Induced Mutation; Kinetics; Lesion; Malignant Neoplasms; Malignant neoplasm of lung; Massive Parallel Sequencing; Measurement; Measures; Medical; Methods; Molecular; Monitor; Mutagenesis; Mutagens; Mutate; Mutation; Mutation Detection; Mutation Fixation; Mutation Spectra; Nuclear; Polymerase; Population; Process; Proliferating; Province; Reporter; Research; Research Design; Risk; Role; Site; Somatic Mutation; Source; Technology; Testing; Time; Woman; adduct; age related; assay development; base; cancer risk; carcinogenesis; carcinogenicity; cooking; design; driving force; environmental mutagens; experience; genome-wide; in vivo; insertion/deletion mutation; insight; lifestyle intervention; next generation sequencing; novel; population based; repaired; research study; stem; theories

PROJECT SUMMARY Somatic mutations are the driving force behind carcinogenesis and other age-related diseases, yet the mechanisms underlying their genesis in humans are ambiguous. This proposal seeks to resolve fundamental processes of nuclear mutagenesis and ascertain the utility of monitoring somatic mutation as a biomarker of individual environmental exposure within a well-characterized population. These objectives are now feasible thanks to the development of a novel mutation detection technology termed CypherSeq, which utilizes circular barcoded templates, rolling circle amplification, and massively parallel sequencing to accurately quantify somatic mutation throughout the genome with unprecedented sensitivity. First, the CypherSeq method will be used to establish a genome-wide profile of somatic mutation in disease-free humans, and test how the frequency, distribution, and spectrum of mutation change with age throughout the genome. Second, proliferating and quiescent human cells will be treated with known mutagens, and induced mutagenesis will be tracked by CypherSeq. These experiments will explore the relationships between cell proliferation, transcription, DNA repair, and somatic mutation, while also uncovering highly mutable regions of the genome, which might serve as sites to monitor acute and chronic mutagen exposure in human populations. Further, this Aim will also test the hypothesis that proliferation is required for repair and mutation fixation within non- transcribed regions of the genome. Lastly, in the third Aim, robust mutational target sites will be characterized and used to monitor mutation frequency in blood draws from individuals who have endured a long-term, carcinogenic exposure to smoky coal. This cross-sectional study will test the hypothesis that underlying the carcinogenicity of smoky coal is its potential to increase the frequency of somatic mutation in exposed individuals. As such, it is expected that increased lifetime exposure to smoky coal will positively correlate with mutation induction. Successful completion of the proposed Aims would provide new insights into human mutagenesis, and highlight the potential utility of monitoring in vivo mutation induction as a biomarker of mutagenic environmental exposure; and thus identify individuals with an elevated risk of developing cancer. Ultimately, mutation-based biomarkers have the potential to stratify cancer risk, providing a basis to direct medical intervention, lifestyle changes (i.e. limiting mutagen exposure), early diagnosis, and/or the application of chemopreventive measures, and thus save lives.

项目概要 体细胞突变是致癌和其他与年龄相关的疾病背后的驱动力，但 它们在人类中的起源机制尚不明确。该提案旨在解决根本问题 核诱变过程并确定监测体细胞突变作为生物标志物的效用 特征明确的人群中的个体环境暴露。这些目标现在是可行的 得益于一种名为 CypherSeq 的新型突变检测技术的开发，该技术利用循环 条形码模板、滚环扩增和大规模并行测序可准确定量 整个基因组的体细胞突变具有前所未有的敏感性。首先，CypherSeq 方法将是 用于建立无病人类体细胞突变的全基因组谱，并测试 整个基因组中突变的频率、分布和谱随着年龄的增长而变化。第二， 增殖和静止的人类细胞将用已知的诱变剂进行处理，并诱导诱变 由 CypherSeq 跟踪。这些实验将探索细胞增殖之间的关系， 转录、DNA 修复和体细胞突变，同时还揭示基因组的高度可变区域， 它可以作为监测人群中急性和慢性诱变剂暴露的场所。此外，这 Aim 还将检验以下假设：非修复和突变固定需要增殖。 基因组的转录区域。最后，在第三个目标中，将表征稳健的突变目标位点 并用于监测从经历了长期、 接触烟煤会致癌。这项横断面研究将检验以下假设： 烟煤的致癌性是其可能增加暴露在外的人体细胞突变的频率 个人。因此，预计一生中接触烟煤的时间增加将与 突变诱导。成功完成拟议目标将为人类提供新的见解 诱变，并强调监测体内突变诱导作为生物标志物的潜在效用 致突变的环境暴露；从而识别出患癌症风险较高的个体。 最终，基于突变的生物标志物有可能对癌症风险进行分层，为指导癌症风险提供基础。 医疗干预、生活方式改变（即限制诱变剂暴露）、早期诊断和/或应用 化学预防措施，从而挽救生命。