DNA Repair Capacity Assays for Lung Disease Risk Assessment

用于肺部疾病风险评估的 DNA 修复能力测定

• 批准号: 10470762
• 负责人: Steven A Belinsky
• 金额: $ 80.61万
• 依托单位: LOVELACE BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10470762
• 关键词: Address; Affect; Automobile Driving; Base Excision Repairs; Biological Assay; Blood; Bronchodilator Agents; Bronchoscopy; CCRL2 gene; Carcinogen exposure; Carcinogens; Case-Control Studies; Cause of Death; Cells; Cessation of life; Charge; Chronic Obstructive Pulmonary Disease; Cigarette; Cigarette Smoker; Clinic; Collaborations; Comet Assay; Comparative Study; Cryopreservation; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Repair Pathway; DNA-dependent protein kinase; Diagnosis; Disease; Disease Progression; Electronic cigarette; Enrollment; Environmental Exposure; Environmental Risk Factor; Epidemiology; Epithelial; Epithelial Cells; Ethnic Origin; Etiology; Event; Excision; Exposure to; Flow Cytometry; Gene Silencing; Genes; Genetic Determinism; Goals; Health; Hispanic; Hospitals; Human; Hydrogen Peroxide; Hypermethylation; IL2 gene; In Vitro; Individual; Individual Differences; Inflammation; Lung; Lung diseases; Lymphocyte; Mainstreaming; Malignant Neoplasms; Malignant neoplasm of lung; Measures; Mediating; Medical; Methylation; Mononuclear; Mutagens; New Mexico; Nonhomologous DNA End Joining; Not Hispanic or Latino; Nucleotide Excision Repair; OGG1 gene; Outcome; Participant; Particulate Matter; Pathogenesis; Performance; Peripheral; Plasmids; Play; Population; Population Study; Predisposition; Proteins; Publications; Quality of life; Questionnaires; Radon; Recording of previous events; Reporter; Research; Resources; Respiratory Disease; Respiratory Signs and Symptoms; Risk; Risk Assessment; Risk Factors; Role; Side; Smoke; Smoker; Smoking; Smoking Status; Spirometry; Sputum; Stream; Testing; Tobacco-Associated Carcinogen; Translating; Update; Variant; Work; base; biomarker development; bronchial epithelium; cancer diagnosis; cancer risk; carcinogenicity; case control; cigarette smoke; cigarette smoking; cohort; demographics; disease phenotype; disorder risk; e-cigarette aerosols; environmental agent; environmental stressor; environmental tobacco smoke; enzyme activity; ethnic difference; experience; fine particles; follow-up; gene environment interaction; gene panel; genome integrity; high risk; homologous recombination; human subject; inter-individual variation; mortality; never smoker; nicotine exposure; oxidative damage; precision medicine; predictive marker; predictive test; promoter; pulmonary function; repaired; respiratory; response; risk prediction; risk variant; second-hand cigarette smoke; study population; wood smoke

Project Summary Lung cancer (LC) is the leading cancer-related cause of death and COPD is third among all cause mortality. There are approximately 70 million current and former cigarette smokers (SM) in the US, although only 15-20% and ~17% will be diagnosed with LC or COPD, respectively. Thus, differences in susceptibility have been proposed to play an important role in LC and COPD disease etiology that also differs between non- Hispanic white (NHW) and Hispanic ethnicity. Smoking is estimated to be the cause of 85% of LC deaths and strongly associated with COPD. Exposure to other environmental respiratory carcinogens that include radon, wood smoke, and particulate matter 2.5 µ (PM2.5) may interact with cigarette smoking or individually be carcinogenic. LC in never smokers (NS) is increasing with 15-20,000 cases annually in the US. Risk factors include environmental tobacco smoke (sidestream smoke is the major component), radon, wood smoke and PM2.5. Wood smoke and PM2.5 are also associated with risk for COPD and most recently electronic cigarettes have been shown to affect pulmonary function. All of these environmental exposures have commonality in their ability to induce DNA damage. Reduced DNA repair capacity (DRC) has been shown to be associated with LC largely through hospital-based case-control studies, but has not been rigorously studied for COPD. These studies used the host cell reactivation or the mutagen sensitivity assay in response to specific DNA damaging agents. A major goal for this RFA “Expanding Genome Integrity Assays to Population Studies” is to develop and/or evaluate the performance of high throughput, functional DNA repair assays in the context of assessing disease phenotype and pathogenesis in a population setting. Our approach to this charge is to evaluate three emerging high throughput DRC assays – Litron in vitro flow cytometry-based MN assay, Trevigen comet assay, and the Trevigen hOGG1 FLARETM comet assay. In addition, through collaboration with Dr. Nagel, the performance of his flow cytometric host cell reactivation assay (FM-HCR) that measures the ability of human cells to repair different types of damage within plasmids transfected into the cells will be assessed. We will take advantage of two established cohorts, the LSC and New Mexico LC cohort to evaluate these DRC assays for predicting LC risk in SM and NS (Aim 1) and COPD in SM (Aim 2) through the exposure of their PMCs to total particulate matter (TPM) from mainstream and sidestream cigarette smoke, wood smoke, electronic cigarette aerosol, and H2O2 (mimic for radon). Aim 3 will address the ability to use the PMCs as a surrogate for risk prediction for LC and COPD by comparing the three DRC assays and exposures in primary bronchial epithelial cells obtained by bronchoscopy and matched PMCs. In addition, the FM-HCR assay will study controls from aims 1 and 2 that show the largest difference in DRC in response to one or more of the exposures to determine if this assay can replicate that outcome. Whether one or more of the specific DNA repair pathways (e.g., base excision) are driving the differences in DRC will also be determined.

项目概要 肺癌 (LC) 是导致癌症相关死亡的主要原因，而慢性阻塞性肺病 (COPD) 在所有原因中排名第三 尽管美国目前和曾经吸烟者 (SM) 大约有 7000 万。 分别只有 15-20% 和约 17% 的人会被诊断为 LC 或 COPD，因此，易感性存在差异。 已被认为在 LC 和 COPD 疾病病因学中发挥重要作用，非慢性阻塞性肺疾病之间也存在差异。 西班牙裔白人 (NHW) 和西班牙裔种族估计是 85% 的 LC 死亡和吸烟的原因。 与慢性阻塞性肺病（COPD）密切相关。 木材烟雾和颗粒物 2.5 µ (PM2.5) 可能与吸烟相互作用或单独产生 在美国，从不吸烟者 (NS) 中的 LC 每年增加 15-20,000 例。 包括环境烟草烟雾（侧流烟雾是主要成分）、氡气、木材烟雾和 PM2.5 和 PM2.5 也与慢性阻塞性肺病和最近的电子病风险相关。 香烟已被证明会影响肺功能。 已证明它们具有诱导 DNA 损伤 (DRC) 能力的共性。 主要通过医院病例对照研究与 LC 相关，但尚未经过严格研究 这些研究使用宿主细胞再激活或诱变剂敏感性测定来响应。 本次 RFA 的主要目标是“将基因组完整性检测扩展到人群”。 研究”是为了开发和/或评估高通量、功能性 DNA 修复检测的性能 我们在人群环境中评估疾病表型和发病机制的背景。 旨在评估三种新兴的高通量 DRC 测定 – Litron 基于体外流式细胞术的 MN 测定， 此外，通过与 Trevigen 彗星测定和 Trevigen hOGG1 FLARETM 彗星测定合作。 Nagel 博士，他的流式细胞术宿主细胞再激活测定 (FM-HCR) 的性能可测量 人类细胞修复转染到细胞中的质粒内不同类型损伤的能力将 我们将利用两个已建立的队列（LSC 和新墨西哥州 LC 队列）进行评估。 这些 DRC 检测通过暴露来预测 SM 和 NS 中的 LC 风险（目标 1）以及 SM 中的 COPD（目标 2） 主流和侧流香烟烟雾、木材烟雾、 电子烟气雾剂和 H2O2（模拟氡气）的目标 3 将解决使用 PMC 作为替代品的能力。 通过比较初级研究中的三种 DRC 检测和暴露来替代 LC 和 COPD 的风险预测 通过支气管镜检查获得的支气管上皮细胞和匹配的 PMC 此外，FM-HCR 测定将进行。 目标 1 和 2 的研究对照显示了 DRC 对一项或多项的反应的最大差异 以确定该测定是否可以复制该结果是否是一种或多种特定 DNA。 驱动 DRC 差异的修复途径（例如碱基切除）也将被确定。