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.

项目摘要 肺癌（LC）是与癌症相关的主要原因，COPD在所有原因中排名第三 死亡。在美国，当前和以前的吸烟者（SM）大约有7000万 只有15-20％和〜17％的LC或COPD诊断。那是易感性的差异 已提出在LC和COPD病病因中发挥重要作用，这在非 - 西班牙裔白人（NHW）和西班牙裔种族。估计吸烟是LC死亡的85％的原因 与COPD密切相关。暴露于其他环境呼吸道癌，包括ra， 木烟以及特定物质2.5 µ（PM2.5）可能与吸烟或单独相互作用 致癌。在美国，LC中的LC（NS）每年在15-20,000例案例中增加。风险因素 包括环境烟草烟雾（横向烟雾是主要成分），ra，木烟和 PM2.5。木烟和PM2.5也与COPD的风险和最近的电子有关 香烟已被证明会影响肺功能。所有这些环境暴露有 它们诱导DNA损伤的能力的共同点。 DNA修复能力降低（DRC）已显示为 主要通过基于医院的病例对照研究与LC相关，但尚未严格研究 对于COPD。这些研究使用了宿主细胞重新激活或诱变者的敏感性评估 特定的DNA损伤剂。这种RFA的主要目标“将基因组完整性分析扩展到人群 研究”是为了开发和/或评估高吞吐量，功能性DNA修复测定法的性能 评估人口环境中疾病表型和发病机理的背景。我们对此指控的方法 是为了评估三个出现的高吞吐量DRC分析 - 基于体外流式细胞仪的LITRON，MN的MN分析， Trevigen Comet分析和Trevigen Hogg1 Flaretm彗星测定法。此外，通过与 Nagel博士，他的流式细胞术宿主细胞重新激活测定法（FM-HCR）的性能 人类细胞修复翻译成细胞内不同类型损伤的能力将是 评估。我们将利用两个已建立的同伙，LSC和New Mexico LC队列进行评估 这些DRC通过曝光通过SM和NS（AIM 1）的LC风险（AIM 1）和COPD（AIM 2）进行的这些DRC测定 从主流和侧面香烟烟，木烟， 电子香烟气溶胶和H2O2（ra的模拟）。 AIM 3将解决将PMC用作的能力 通过比较初级的三种DRC分析和暴露，以替代LC和COPD的风险预测 支气管上皮细胞通过支气管镜检查和匹配的PMC获得。此外，FM-HCR分析将 AIM 1和2的研究对照显示了drc的最大差异，以响应一个或多个 暴露以确定该测定法是否可以复制该结果。是一个或多个特定的DNA 维修途径（例如，基本惊喜）也将确定刚果民主共和国的差异。