TGF-Beta Regulates CFTR-Mediated Airway Smooth Muscle Dysfunction

项目总结/摘要 囊性纤维化（CF）是一种致命的遗传性疾病，其特征是进行性肺部疾病和气道阻塞。 CF是由CF跨膜传导调节因子（CFTR）突变引起的。CF患者经常 发生与平滑肌功能障碍相关的气道高反应性（AHR）， 阻塞并预示着更快的肺功能下降。TGF β 1是CF的遗传修饰剂， 与更严重的肺部疾病有关TGF β 1还导致CF小鼠AHR和平滑肌异常恶化 对比非CF小鼠。目前尚不清楚TGF β 1如何改变CF肺病或驱动CF平滑肌功能障碍。 该建议旨在研究TGF β 1在CF气道平滑肌功能障碍中的作用。目前，没有治疗方法 直接靶向CF中的平滑肌异常或TGF β 1信号传导。本提案调查了 TGF β 1通过影响CF和CFTR调节CFTR介导的气道平滑肌功能障碍的假说 上皮细胞和平滑肌细胞。CF动物和小鼠/人原代细胞培养模型将用于测试 CFTR在TGF β诱导的肺疾病中的组织特异性功能。目标1将决定CFTR的作用 气道上皮细胞功能障碍介导TGF β-驱动的肺部疾病，使用TGF β-暴露的上皮细胞， 特异性Cftr敲除小鼠和原代鼠和人细胞培养模型。这些研究将侧重于 上皮和炎症介质。目的2探讨TGF β 1诱导CF气道通畅的机制 肌肉功能障碍，通过检查肺和平滑肌异常，在平滑肌特异性 Cftr敲除小鼠和分离的鼠和人CF细胞培养模型。TGF β介导的AHR试验，肺 功能，并且平滑肌收缩性将用于比较CF和非CF气道平滑肌功能。 本提案的主要研究者伊丽莎白·克雷默博士是肺部医学领域的一名医师科学家，重点研究 早期CF肺病的驱动因素。她有博士学位在分子和发育生物学和广泛的 小鼠模型和肺生理学的背景。她的导师提供CF鼠标方面的免费专业知识 和细胞培养模型（A. P. Naren博士，主要导师），转化CF研究和个性化医疗 (Dr. J. P. Clancy，共同导师）和转化研究设计（Raouf Amin博士，共同导师）。这些导师 在指导学术科学家方面有成功的记录。他们个人致力于博士。 克雷默的成功完成这一建议，并过渡到一个独立的研究生涯。 本申请表中列出的培训计划将为申请人提供创新实验室的关键培训 技术，尖端的生物信息学分析，免疫学和先进的职业技能，成功地 建立一个独立的翻译科学计划。这个培训计划利用了优秀的 辛辛那提儿童医院的环境和支持。实现这些培训目标并完成 本申请中描述的研究将建立在克雷默博士先前在分子和发育方面的专业知识基础上 生物学，提供了一个坚实的科学和培训平台，开始她的独立研究生涯。

PROJECT SUMMARY/ABSTRACT Cystic fibrosis (CF) is a lethal genetic disorder characterized by progressive lung disease and airway obstruction. CF is caused by mutations in the CF transmembrane conductance regulator (CFTR). Patients with CF often develop airway hyperresponsiveness (AHR) related to smooth muscle dysfunction, which worsens airway obstruction and portends faster lung function decline. TGF is a genetic modifier of CF, with higher TGF levels linked to more severe lung disease. TGF also causes worse AHR and smooth muscle abnormalities in CF mice versus non-CF mice. It is unknown how TGF modifies CF lung disease or drives CF smooth muscle dysfunction. This proposal seeks to investigate TGF’s role in CF airway smooth muscle dysfunction. Currently, no therapies directly target smooth muscle abnormalities or TGF signaling in CF. This proposal investigates the overall hypothesis that TGF regulates CFTR-mediated airway smooth muscle dysfunction through effects on both CF epithelial and smooth muscle cells. CF animal and mouse/human primary cell culture models will be used to test the tissue specific function of CFTR in TGF induced lung disease. Aim 1 will determine the role of CFTR dysfunction in airway epithelial cells in mediating TGF-driven lung disease, using TGF exposed epithelial- specific Cftr knockout mice and primary murine and human cell culture models. These studies will focus on epithelial and inflammatory mediators. Aim 2 will test the mechanisms of TGF-induced CF airway smooth muscle dysfunction by examining pulmonary and smooth muscle abnormalities in both smooth muscle-specific Cftr knockout mice and isolated murine and human CF cell culture models. Tests of TGF-mediated AHR, lung function, and smooth muscle contractility will be used to compare CF and non-CF airway smooth muscle function. The PI for this proposal, Dr. Elizabeth Kramer, is a physician scientist in Pulmonary Medicine with a focus on drivers of early CF lung disease. She has a Ph.D. in Molecular and Developmental Biology and an extensive background in mouse models and lung physiology. Her mentors provide complimentary expertise in CF mouse and cell culture models (Dr. A.P. Naren, primary mentor), translational CF research and personalized medicine (Dr. J.P. Clancy, Co-mentor), and translational research design (Dr. Raouf Amin, Co-mentor). These mentors have an established record of success in mentoring academic scientists. They are personally committed to Dr. Kramer’s success in completing this proposal and transitioning to an independent research career. The training plan outlined in this application will provide the applicant with crucial training in innovative laboratory techniques, cutting-edge bioinformatics analysis, immunology, and advanced career skills to successfully establish an independent translational science program. This training plan capitalizes on the excellent environment and support at Cincinnati Children’s Hospital. Addressing these training goals and completing the studies described in this application will build upon Dr. Kramer’s prior expertise in molecular and developmental biology, providing a solid scientific and training platform to launch her independent research career.