Role of Werner's Syndrome Protein in Cigarette Smoke-Induced Cellular Senescence

维尔纳综合征蛋白在香烟烟雾诱导的细胞衰老中的作用

基本信息

批准号：
7468711
负责人：
Toru Nyunoya
金额：
$ 12.64万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-01 至 2013-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7468711
关键词：
Age Aging Area Award Biology of Aging Biometry Cell Aging Cell Cycle Arrest Cell model Cells Cellular Morphology Cellular biology Chronic Chronic Obstructive Airway Disease Cigarette DNA Damage Data Development Development Plans Disease Down-Regulation Ensure Environment Epithelial Cells Exposure to Extracellular Matrix Family Fibroblasts Functional disorder Future Galactosidase Genes Genetic Genomic Instability Habits Hereditary Disease Human Impaired wound healing In Situ In Vitro Investigation Link Lung Lung diseases Mediating Mentors Modality Modeling Molecular Mutation Pathogenesis Pathway interactions Patients Phenotype Physicians Plasmids Premature aging syndrome Protein Overexpression Proteins Publishing Pulmonary Emphysema Rate Regulation Research Research Design Research Personnel Retinoblastoma Protein Role Scientist Smoke Smoker Smoking Structure of parenchyma of lung TP53 gene Training Transfection WRN gene Werner Syndrome Wound Healing abstracting career cell motility cigarette smoke-induced cigarette smoking cigarette smoking helicase human WRN protein in vitro Model loss of function mutation lung injury member migration multidisciplinary novel protein expression response senescence small hairpin RNA success

项目摘要

DESCRIPTION (provided by applicant): This application is to support the development of Dr. Toru Nyunoya into an independent researcher and an accomplished physician-scientist. Dr. Gary Hunninghake will support him as primary mentor to ensure the success of his career development plan. Specific areas of career development during this award include coursework, including cell biology, genetics, and biostatistics. The focus of this application is the investigation of mechanisms of cigarette smoke-induced fibroblast senescence. These studies directly relate to the pathogenesis of chronic obstructive pulmonary disease (COPD), which has been linked to cellular senescence, accelerated aging and impaired wound repair. Werner's syndrome (WS), a genetic disorder caused by loss-of-function mutations in the Werner's syndrome gene encoding a member of RecQ helicase family (WRN protein) also accelerates aging. However, to date, no study has explored the potential link between cigarette smoke-induced accelerated aging and WS. The candidate hypothesis is that exposure to cigarette smoke induces fibroblast senescence via WRN protein downregulation. In Aim 1, Dr. Nyunoya will identify the molecular pathway(s) responsible for cigarette smoke-induced cellular senescence in lung fibroblasts and in circulating fibrocytes from patients with COPD. Cigarette smoke-induced alterations in WRN protein expression will be examined in these different human models. To counter the cigarette smoke effect, WRN protein will be overexpressed using WRN plasmid transfection in cultured fibroblasts. A potential link between WRN protein and known senescence-inducing pathways such as p53 and p16 will be explored by knockdown using short hairpin RNA. In Aim 2, Dr. Nyunoya will determine the role of cigarette smoke-induced cellular senescence in an in vitro model of wound healing. Specifically, the role of p53, p16 and WRN protein in cigarette smoke-induced migration dysfunction and extracellular matrix regulation will be determined by shRNA knockdown of p53 and p16 or overexpression of WRN protein. Throughout Dr. Nyunoya's career development, he will meet with advisors in cell biology, aging, biostatistics and study design. This multidisciplinary training environment will enhance his career development and ensure that he evolves into an independent researcher. Elucidating the mechanisms of cigarette smoke-induced fibroblast senescence will contribute to our understanding of COPD pathogenesis and generate direction for future treatment modalities. (End of Abstract)

描述（由申请人提供）：此应用程序旨在支持 Toru Nyunoya 博士发展成为一名独立研究员和一名卓有成就的医师科学家。 Gary Hunninghake 博士将作为主要导师支持他，以确保他的职业发展计划取得成功。该奖项期间职业发展的具体领域包括课程作业，包括细胞生物学、遗传学和生物统计学。该应用的重点是研究香烟烟雾诱导成纤维细胞衰老的机制。这些研究与慢性阻塞性肺病（COPD）的发病机制直接相关，慢性阻塞性肺病与细胞衰老、加速衰老和伤口修复受损有关。沃纳氏综合症 (WS) 是一种由编码 RecQ 解旋酶家族成员（WRN 蛋白）的沃纳氏综合症基因功能丧失突变引起的遗传性疾病，也会加速衰老。然而，迄今为止，还没有研究探讨香烟烟雾引起的加速衰老与 WS 之间的潜在联系。候选假设是，接触香烟烟雾会通过 WRN 蛋白下调诱导成纤维细胞衰老。在目标 1 中，Nyunoya 博士将确定导致慢性阻塞性肺病患者肺成纤维细胞和循环纤维细胞中香烟烟雾诱导细胞衰老的分子途径。将在这些不同的人类模型中检查香烟烟雾引起的 WRN 蛋白表达的变化。为了对抗香烟烟雾的影响，将使用 WRN 质粒转染在培养的成纤维细胞中过度表达 WRN 蛋白。将通过使用短发夹 RNA 进行敲低来探索 WRN 蛋白与已知的衰老诱导途径（例如 p53 和 p16）之间的潜在联系。在目标 2 中，Nyunoya 博士将确定香烟烟雾诱导的细胞衰老在体外伤口愈合模型中的作用。具体来说，p53、p16 和 WRN 蛋白在香烟烟雾诱导的迁移功能障碍和细胞外基质调节中的作用将由 p53 和 p16 的 shRNA 敲低或 WRN 蛋白的过表达来决定。在 Nyunoya 博士的职业发展过程中，他将与细胞生物学、衰老、生物统计学和研究设计方面的顾问会面。这种多学科的培训环境将促进他的职业发展，并确保他发展成为一名独立的研究员。阐明香烟烟雾诱导成纤维细胞衰老的机制将有助于我们了解慢性阻塞性肺病的发病机制，并为未来的治疗方式提供方向。（摘要完）