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: 9768991
• 负责人: Steven A Belinsky
• 金额: $ 81.21万
• 依托单位: LOVELACE BIOMEDICAL & ENVIRONMENTAL 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 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9768991
• 关键词: Address; Aerosols; 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 Airway 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; Environmental Tobacco Smoke; Epidemiology; Epithelial Cells; Epithelium; Ethnic Origin; Etiology; Event; Excision; Exposure to; Flow Cytometry; Gene Silencing; Genes; Genetic Determinism; Goals; Health; Hispanics; 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 Signs and Symptoms; Respiratory physiology; 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; cancer diagnosis; cancer risk; carcinogenicity; case control; cigarette smoke; cigarette smoking; cohort; demographics; disease phenotype; disorder risk; environmental agent; environmental stressor; enzyme activity; ethnic difference; experience; 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 variant; study population; vapor; 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（Lung cancer，LC）是导致死亡的主要癌症相关原因，COPD是所有原因中的第三位 mortality.在美国，大约有7000万现在和以前的吸烟者（SM），尽管 只有15-20%和~17%分别被诊断为LC或COPD。因此，易感性的差异 已经提出在LC和COPD疾病病因学中起重要作用，这也与非 西班牙裔白色（NHW）和西班牙裔。据估计，吸烟是85%的LC死亡的原因， 与COPD密切相关。暴露于其他环境呼吸致癌物，包括氡， 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 致癌的在美国，从不吸烟者（NS）中的LC每年增加15- 20，000例。危险因素 包括环境烟草烟雾（侧流烟雾是主要成分）、氡、木材烟雾和 PM2.5。木材烟雾和PM2.5也与COPD的风险有关，最近的电子 香烟已被证明会影响肺功能。所有这些环境暴露 它们在诱导DNA损伤的能力上的共性。DNA修复能力（DRC）降低 通过以医院为基础的病例对照研究，大部分与LC相关，但尚未进行严格研究 慢性阻塞性肺病这些研究使用宿主细胞再活化或诱变剂敏感性试验， 特定的DNA损伤剂RFA的一个主要目标是“将基因组完整性检测扩展到人群 研究”是开发和/或评估高通量、功能性DNA修复测定在 在人群环境中评估疾病表型和发病机制的背景下。我们对这项指控的态度 是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， Trevigen彗星试验和Trevigen hOGG 1 FLARETM彗星试验。此外，通过与 博士内格尔，他的流式细胞术宿主细胞再活化试验（FM-HCR）的性能，测量 人细胞修复转染入细胞的质粒内不同类型损伤的能力将被 评估。我们将利用两个已建立的队列，LSC和新墨西哥州LC队列， 这些DRC检测通过暴露预测SM和NS（目的1）中的LC风险和SM（目的2）中的COPD风险 从主流和侧流香烟烟雾，木材烟雾， 电子香烟气溶胶和H2 O2（模拟氡）。目标3将解决使用私营军事公司作为 通过比较三种DRC检测和主要受试者的暴露量， 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检测试剂盒将 目标1和2的研究对照，显示对一种或多种 以确定该检测是否可以复制该结果。是否有一个或多个特定的DNA 修复途径（例如，碱基切除）驱动DRC的差异也将被确定。