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

描述（由申请人提供）：本次K01指导职业奖申请的目标是通过研究经验，学术活动，培训和合作努力来推进Roan博士的职业生涯。这将使她能够在孟菲斯大学（U of M）建立一个完善的生物医学研究项目，研究肺部健康和生理学，这是一个城市环境中的多元化机构。更具体地说，Roan博士将通过一系列研究肺泡渗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，这将使他从一个具有软组织力学和力学生物学背景的研究者转变为一个专注于急性肺损伤的独立研究者。提出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，部分特征是肺泡空气中存在富含蛋白质的水肿。由于水肿的清除对疾病的解决至关重要，因此需要针对保护上皮屏障和促进上皮修复的策略。中心假设是肿瘤坏死因子-α (TNF-α)诱导肺泡上皮细胞内细胞骨架张力增加，使它们更容易受到屏障功能障碍和细胞脱离的影响，这是通过c-Jun NH2激酶- (JNK-)介导的闭塞带1 （ZO-1）磷酸化引起的。在Aim 1中，Roan博士将证明，由于细胞硬度增加，TNF-α和拉伸会导致比单独刺激更多的细胞脱离。Aim 2将表明僵硬度的增加和屏障功能障碍依赖于通过JNK磷酸化ZO-1。在Aim 3中，将开发一个多尺度有限元模型来预测TNF-α引起的关节应力变化如何导致体内屏障功能障碍增加。总体而言，该项目将阐明机械通气前肺泡上皮细胞早期暴露于TNF-α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。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. Waters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工作。培训活动将在孟菲斯大学和UTHSC进行，这两所合作院校相距约10分钟车程，并通过一系列校际活动协同开展生物医学研究。UofM同意分配Roan博士75%的成果，UofM和UTHSC都同意提供所需的课程和实验室设施，以开展拟议的5年指导研究和职业发展项目，帮助Roan博士推进她的职业生涯，并在mm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.