TNF-a Induced Increase in Alveolar Epithelial Cell Stiffness Increase and Lung In

描述(由申请者提供)：本次K01职业指导奖申请的目标是通过研究经验、学术活动、培训和合作努力推进Roan博士的职业生涯。这将使她能够在孟菲斯大学(U Of M)建立一个健全的肺部健康和生理学生物医学研究计划，这是一个城市环境中的多元化机构。更具体地说，罗恩博士将通过一系列强有力的研究肺泡通透性的活动来发展肺生理学和气液屏障功能方面的专业知识，这些活动将使具有软组织力学和机械生物学背景的研究人员转变为专注于急性肺损伤的独立研究人员。拟议的研究重点是急性呼吸窘迫综合征，这是一种破坏性的疾病，部分特征是肺泡内存在富含蛋白质的浮肿。由于清除水肿对疾病的解决至关重要，因此需要有针对性地保护上皮屏障和促进上皮修复的策略。中心假说是肿瘤坏死因子-α(Tumor NectorFor-α，肿瘤坏死因子-JNK)诱导肺泡上皮细胞内细胞骨架张力增加，使其更容易受到通过c-jun NH2激酶(JNK-)介导的闭锁带1(ZO-1)的磷酸化而引起的屏障功能障碍和细胞脱离的影响。在目标1中，罗恩博士将证明，由于细胞僵硬的增加，肿瘤坏死因子-α和拉伸将比单独使用任一种刺激导致更多的细胞分离。目的2将证明僵硬的增加和屏障功能障碍依赖于ZO-1通过JNK的磷酸化。在目标3中，将开发一个多尺度有限元模型来预测由肿瘤坏死因子-α引起的连接应力的变化如何导致体内屏障功能障碍的增加。总体而言，本项目将阐明在机械通气前早期将肺泡上皮细胞暴露于肿瘤坏死因子-α是如何诱导细胞骨架重塑并改变紧密连接处的生物物理力从而导致更多损伤的。候选人将由知名研究人员指导，田纳西大学健康科学中心(UTHSC)生理学教授兼副主席C.M.沃特斯博士担任首席导师。R.K.Rao博士和E.Fitzpatrick博士将分别指导紧密连接生物学和免疫学方面的研究。培训活动将在孟菲斯大学和UTHSC进行，这两个合作机构相距大约10分钟的车程，并通过一系列校际活动协同开展生物医学研究。密歇根大学同意为罗恩博士的工作分配75%的经费，密歇根大学和密歇根州立大学HSC都同意根据需要提供课程和实验室设施，为罗恩博士开展拟议的为期5年的指导性研究和职业发展项目，以推进她的职业生涯，并在密歇根大学建立一个健全的研究项目。

DESCRIPTION (provided by applicant): The goal of this K01 Mentored Career Award application is to advance Dr. Roan's career through research experiences, scholarly activities, training, and collaborative efforts. This will enable her to establish a sound Biomedical Research Program in Lung Health and Physiology at the University of Memphis (U of M), a diverse institution in an urban environment. More specifically, Dr. Roan will develop expertise in lung physiology and air-fluid barrier functionality through a robust set of activities that investigate alveolar permeability that will enable transition from an investigator with a background in soft tissue mechanics and mechanobiology to an independent researcher focused on acute lung injury. The proposed research focuses on acute respiratory distress syndrome, a devastating condition characterized in part by the presence of protein rich edema in the alveolar airspaces. Because clearance of edema is critical to resolution of the disease, strategies directed toward protecting the epithelial barrier and promoting epithelial repair are needed. The central hypothesis is that tumor necrosis factor-α (TNF-α) induces an increase in cytoskeletal tension within the alveolar epithelial cells making them more susceptible to barrier dysfunction and cell detachment caused by stretch through c-Jun NH2 kinase- (JNK-) mediated phosphorylation of zonula occludens 1 (ZO-1). In Aim 1, Dr. Roan will show that TNF-α and stretch will lead to much more cell detachment than either stimulus alone due to an increase in cell stiffness. Aim 2 will show that the increase in stiffness and barrier dysfunction are dependent on ZO-1 phosphorylation through JNK. In Aim 3, a multi- scale finite element model will be developed to predict how changes in junctional stress caused by TNF-α can lead to increased barrier dysfunction in vivo. Overall, this project will elucidate how early exposure of alveolar epithelial cells to TNF-α prior to mechanical ventilation induces cytoskeletal remodeling and alters the biophysical forces at the tight junctions leading to more injury. The candidate will be mentored by well- established investigators, with Dr. C.M. Waters, Professor and Vice Chair, Physiology, University of Tennessee Health Science Center (UTHSC) serving as the lead mentor. Dr. R.K. Rao and Dr. E. Fitzpatrick will guide the tight-junction biology and immunology aspects, respectively. Training activities will take place at both the University of Memphis and the UTHSC, partner institutions separated by an approximately 10 minute drive and synergistically pursuing biomedical research through a range of inter-campus activities. The UofM has agreed to allocate 75% of Dr. Roan's effort, and both the UofM and the UTHSC have agreed to provide access to curricular and laboratory facilities as needed to carry out the proposed 5-year mentored research and career development project for Dr. Roan to advance her career and establish a sound research program here at the U of M.