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

描述（由申请人提供）： 此应用程序是为了支持Toru Nyunoya博士发展成为一名独立的研究人员和一名有成就的医生科学家。加里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。在此期间，职业发展的具体领域包括课程，包括细胞生物学，遗传学和生物统计学。本申请的重点是研究香烟烟雾诱导的成纤维细胞衰老的机制。这些研究直接涉及慢性阻塞性肺疾病（COPD）的发病机制，这与细胞衰老，加速老化和受损的伤口修复有关。沃纳综合征（WS），一种由编码RecQ解旋酶家族（WRN蛋白）成员的沃纳综合征基因中的功能丧失突变引起的遗传性疾病，也加速衰老。然而，到目前为止，还没有研究探讨香烟烟雾诱导的加速衰老和WS之间的潜在联系。候选假设是，暴露于香烟烟雾诱导成纤维细胞衰老通过WRN蛋白下调。在目标1中，Nyunoya博士将确定COPD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诱导的WRN蛋白表达的改变。为了对抗香烟烟雾效应，将使用WRN质粒转染在培养的成纤维细胞中过表达WRN蛋白。WRN蛋白和已知的衰老诱导途径如p53和p16之间的潜在联系将通过使用短发夹RNA敲低来探索。在目标2中，Nyunoya博士将确定香烟烟雾诱导的细胞衰老在伤口愈合的体外模型中的作用。具体而言，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通过p53和p16的shRNA敲低或WRN蛋白的过表达来确定。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人员。阐明香烟烟雾诱导成纤维细胞衰老的机制将有助于我们对COPD发病机制的理解，并为未来的治疗方法提供方向。(End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)