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：
7779961
负责人：
Toru Nyunoya
金额：
$ 5.83万
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 2010-08-03
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7779961
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 Proteins Publishing Pulmonary Emphysema Regulation Research Research Design Research Personnel Retinoblastoma Protein Role Scientist Smoke Smoker Structure of parenchyma of lung TP53 gene Training Transfection Werner Syndrome Wound Healing abstracting career development cell motility cigarette smoke-induced cigarette smoking cigarette smoking helicase human WRN protein in vitro Model loss of function mutation lung injury meetings member migration multidisciplinary novel overexpression premature 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）：该应用是为了支持Toru Nyunoya博士的发展成一个独立的研究人员和一位出色的医师科学家。 Gary Hunninghake博士将支持他作为主要导师，以确保他的职业发展计划的成功。该奖项期间职业发展的特定领域包括课程，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的成纤维细胞衰老机制。这些研究与慢性阻塞性肺疾病（COPD）的发病机理直接相关，该发病机理与细胞衰老，加速衰老和伤口修复受损有关。 Werner's Syndrome（WS）是由编码RECQ解旋酶家族成员（WRN蛋白）的Werner综合征基因丧失功能突变引起的遗传疾病，也可以加速衰老。但是，迄今为止，尚无研究探索烟雾引起的加速衰老与WS之间的潜在联系。候选假设是，暴露于香烟烟雾会通过WRN蛋白下调诱导成纤维细胞衰老。在AIM 1中，Nyunoya博士将确定负责烟雾诱导的肺成纤维细胞细胞衰老的分子途径以及COPD患者的循环纤维细胞中的细胞衰老。在这些不同的人类模型中，将检查香烟烟雾诱导的WRN蛋白表达的改变。为了应对香烟的烟雾作用，使用培养的成纤维细胞中的WRN质粒转染将过表达WRN蛋白。使用短发夹RNA敲低探索WRN蛋白与已知衰老诱导途径（如p53和p16）之间的潜在联系。在AIM 2中，Nyunoya博士将确定烟雾诱导的细胞衰老在伤口愈合模型中的作用。具体而言，p53，p16和WRN蛋白在烟雾诱导的迁移功能障碍和细胞外基质调控中的作用将由p53和p16的shRNA敲低或WRN蛋白过表达来确定。在Nyunoya博士的职业发展中，他将与细胞生物学，衰老，生物统计学和研究设计的顾问会面。这种多学科的培训环境将增强他的职业发展，并确保他发展成为独立的研究人员。阐明香烟烟雾诱导的成纤维细胞衰老的机制将有助于我们对COPD发病机理的理解，并为将来的治疗方式产生方向。（抽象的结尾）