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

基本信息

批准号：
10333369
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
项目类别：
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/10333369
关键词：
Address Adrenergic beta-Agonists Airway Resistance Animals Automobile Driving Bioinformatics Biological Assay Bronchial Spasm Calcium Caring Cell Culture Techniques Cell model Child Health Clinical Coupled Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Data Defect Development Developmental Biology Disease Doctor of Philosophy Dose Environment Epithelial Epithelial Cells 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 Paracrine Communication Pathogenesis Pathologic Pathology Pathway interactions Pediatric Hospitals Persons Phenotype Physicians Physiology Positioning Attribute Primary Cell Cultures Production Professional Competence 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 base 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

项目摘要

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（CF）是一种致命的遗传疾病，其特征是进行性肺部疾病和气道阻塞。 CF是由CF跨膜电导调节剂（CFTR）突变引起的。 CF患者经常发展与平滑肌功能障碍有关的气道高反应性（AHR），这会使气道恶化阻塞和预示更快的肺功能下降。 TGF是CF的遗传修饰符，具有较高的TGF水平与更严重的肺部疾病有关。 TGF还会导致CF小鼠的AHR和平滑肌异常与非CF小鼠相比。尚不清楚TGF如何修饰CF肺部疾病或驱动CF平滑肌功能障碍。该建议旨在研究TGF在CF气道平滑肌功能障碍中的作用。目前，没有疗法直接靶向CF中的平滑肌异常或TGF信号传导。该提案调查了总体 TGF通过对两种CF的影响调节CFTR介导的气道平滑肌功能障碍的假设上皮和平滑肌细胞。 CF动物和小鼠/人类原代细胞培养模型将用于测试 CFTR在TGF诱导肺部疾病中的组织特异性功能。 AIM 1将决定CFTR的作用使用TGF暴露于TGF 特定的CFTR基因敲除小鼠，原代鼠和人类细胞培养模型。这些研究将重点上皮和炎症介质。 AIM 2将测试TGF诱导的CF气道的机理通过检查两个平滑肌特异性的肺部和平滑肌异常，肌肉功能障碍 CFTR敲除小鼠以及分离的鼠和人类CF细胞培养模型。 TGF介导的AHR测试，肺功能和平滑肌收缩力将用于比较CF和非CF气道平滑肌功能。该建议的PI，伊丽莎白·克莱默（Elizabeth Kramer）博士是肺部医学的物理科学家，重点是早期CF肺部疾病的驱动因素。她有博士学位在分子和发育生物学以及广泛的生物学小鼠模型和肺部生理的背景。她的导师提供了CF鼠标的免费专业知识和细胞培养模型（主要导师A.P. Naren博士），翻译了CF研究和个性化医学（J.P. Clancy博士，联合学）和翻译研究设计（Raouf Amin博士，同事）。这些导师在心理学术科学家中拥有成功的记录。他们亲自致力于博士克莱默（Kramer）成功完成了这项建议，并过渡到独立研究职业。本申请中概述的培训计划将为应用程序提供创新实验室的关键培训技术，尖端的生物信息学分析，免疫学和高级职业技能建立独立的翻译科学计划。该培训计划利用了优秀辛辛那提儿童医院的环境和支持。解决这些培训目标并完成本应用中描述的研究将基于克莱默博士的先前分子和发展专家生物学，提供了一个扎实的科学和培训平台来启动她独立的研究职业。