TGF-Beta Regulates CFTR-Mediated Airway Smooth Muscle Dysfunction

TGF-β 调节 CFTR 介导的气道平滑肌功能障碍

• 批准号: 10549342
• 负责人: Elizabeth L Kramer
• 金额: $ 16.42万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10549342
• 关键词: Address; Adrenergic beta-Agonists; Animals; Automobile Driving; Bioinformatics; Biological Assay; Bronchial Spasm; Calcium; Career Mobility; Caring; Cell Culture Techniques; Cell model; Child Health; Clinical; Coculture Techniques; Coupled; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Defect; Development; Developmental Biology; Disease; Doctor of Philosophy; Dose; Environment; Epithelial Cells; Epithelium; Etiology; Exposure to; Functional disorder; Genes; Genetic; Genetic Diseases; Genetic Polymorphism; Goals; Grant; Human; Immunology; In Vitro; Infection; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; K-Series Research Career Programs; Knockout Mice; Laboratories; Link; Lung; Lung diseases; Mechanics; Mediating; Mentors; Modeling; Molecular Biology; Molecular Biology Techniques; Molecular and Cellular Biology; Morphology; Mus; Muscle function; Mutation; PIK3CG gene; Paracrine Communication; Pathogenesis; Pathologic; Pathology; Pathway interactions; Pediatric Hospitals; Persons; Phenotype; Physicians; Physiology; Positioning Attribute; Primary Cell Cultures; Production; Professional Competence; Proliferating; Proteins; Publishing; Pulmonary Cystic Fibrosis; Pulmonary Pathology; Pulmonology; Reporting; Research; Research Design; Research Personnel; Resistance; Role; Science; Scientist; Severities; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Solid; Stimulus; Techniques; Testing; Tissues; Training; Transforming Growth Factor beta; Transgenic Mice; Translational Research; Writing; airway epithelium; airway hyperresponsiveness; airway obstruction; career; children with cystic fibrosis; cystic fibrosis airway; cystic fibrosis airway epithelia; cystic fibrosis mouse; cystic fibrosis patients; cytokine; early cystic fibrosis; experimental study; functional loss; improved; in vivo; inhibitor; innovation; mouse model; new therapeutic target; novel; obstructive airway disease; paracrine; personalized medicine; programs; pulmonary function; pulmonary function decline; respiratory smooth muscle; response; success; targeted treatment; transcriptome sequencing; translational study

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.

项目摘要/摘要 囊性纤维化是一种以进行性肺部疾病和呼吸道阻塞为特征的致死性遗传性疾病。 Cf是由Cf跨膜电导调节因子(CFTR)突变引起的。慢性纤维性心脏病患者经常 发生与平滑肌功能障碍相关的气道高反应性(AHR)，这会加重呼吸道状况 梗阻，预示肺功能下降更快。转化生长因子是CF的遗传修饰物，具有较高的转化生长因子水平 与更严重的肺部疾病有关。转化生长因子还可导致CF小鼠更严重的AHR和平滑肌异常 与非CF小鼠进行比较。目前尚不清楚转化生长因子是如何改变CF肺部疾病或导致CF平滑肌功能障碍的。 本研究旨在探讨转化生长因子在肺间质纤维化中的作用。目前，还没有治疗方法 直接靶向CF中的平滑肌异常或转化生长因子信号。这项提案调查了整个 转化生长因子通过影响双侧肺间质纤维调节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将使用动物和小鼠/人原代细胞培养模型来测试 成纤维细胞生长因子受体在转化生长因子所致肺部疾病中的组织特异性功能目标1将确定CFTR的作用 在介导转化生长因子驱动的肺部疾病中，呼吸道上皮细胞的功能障碍，使用转化生长因子暴露的上皮细胞。 特定的cftr基因敲除小鼠和原代小鼠和人类细胞培养模型。这些研究将集中在 上皮性和炎性介质。目的2研究转化生长因子诱导的肺间质纤维化的作用机制 通过检查两种平滑肌特异性的肺和平滑肌异常来研究肌肉功能障碍 Cftr基因敲除小鼠和分离的小鼠和人的cf细胞培养模型。转化生长因子介导的促肾上腺皮质激素释放激素受体的检测 功能和平滑肌收缩功能将被用来比较CF和非CF的气道平滑肌功能。 这项提议的私人投资者伊丽莎白·克莱默博士是一位肺部医学的内科科学家，专注于 早期慢性肺病的驱动因素。她拥有分子和发育生物学博士学位，并拥有广泛的 小鼠模型和肺生理学的研究背景。她的导师为她提供了免费的CF鼠标专业知识 和细胞培养模型(A.P.Naren博士，主要导师)、翻译的CF研究和个性化医学 (J.P.Clancy博士，共同导师)和翻译研究设计(Raouf Amin博士，共同导师)。这些导师 在指导学术科学家方面有既定的成功记录。他们个人致力于Dr。 克莱默成功地完成了这项提议，并过渡到独立的研究生涯。 申请书中概述的培训计划将为申请者提供关键的创新实验室培训。 技术、尖端生物信息学分析、免疫学和高级职业技能 建立一个独立的翻译科学项目。这项培训计划充分利用了优秀的 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莱默博士在分子和发育方面的先前专业知识的基础上 生物学，为她开始自主研究生涯提供了坚实的科学和培训平台。