Cholesterol Regulation of EGFR-dependent Vasoconstriction in Chronic Hypoxia-induced Pulmonary Hypertension

慢性缺氧引起的肺动脉高压中 EGFR 依赖性血管收缩的胆固醇调节

基本信息

批准号：
10386244
负责人：
Rosstin Ahmadian
金额：
$ 3.81万
依托单位：
UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10386244
关键词：
Acute Address Affect Altitude Blood Blood Vessels Calcium Cell membrane Cell physiology Cholesterol Chronic Chronic Bronchitis Chronic Obstructive Pulmonary Disease Clinical Communication Coupling Data Development Disease Distal Edema Environment Epidermal Growth Factor Receptor Etiology Goals Human Hypoxia In Vitro Knowledge Lead Lung Mediating Mediator of activation protein Membrane Mentors Mission Modeling Molecular Morbidity - disease rate National Heart, Lung, and Blood Institute Oral Outcome Pathogenesis Pathway interactions Patients Peripheral Phenotype Physicians Physiology Preparation Process Production Protocols documentation Pulmonary Emphysema Pulmonary Hypertension Pulmonary Vascular Resistance Pulmonary artery structure Rattus Reactive Oxygen Species Receptor Signaling Regulation Research Research Training Scientist Signal Pathway Signal Transduction Sleep Apnea Syndromes Smooth Muscle Myocytes Source Stimulus Testing Training Vasoconstrictor Agents Video Microscopy arterial remodeling cell type cellular imaging constriction experience hypoxia-induced pulmonary hypertension imaging study in vivo lung hypoxia medical schools mortality new therapeutic target novel oxidation pressure pulmonary vasoconstriction receptor response right ventricular failure skills vasoconstriction

项目摘要

Project Summary Chronic Hypoxia (CH)-induced pulmonary hypertension (PH) is a significant source of morbidity and mortality in patients with chronic obstructive pulmonary diseases. It is widely recognized that vasoconstriction is a critical mediator of PH, although the mechanisms involved are poorly understood. Our previous studies have demonstrated that enhanced vasoconstrictor sensitivity following CH involves a requisite reduction in pulmonary arterial smooth muscle cell (PASMC) membrane cholesterol content. We have also demonstrated that CH augments vasoconstrictor reactivity by a switch in signaling from primarily calcium-dependent mechanisms to a Ca2+ sensitization pathway that involves the epidermal growth factor receptor (EGFR) and reactive oxygen species (ROS). However, the mechanisms by which CH decreases membrane cholesterol and how this unmasks EGFR-dependent vasoconstriction has yet to be assessed. The proposed studies will investigate the central hypothesis that coupling of vasoconstrictor stimuli to EGFR signaling following CH promotes PASMC hypercontractility through a ROS-dependent decrease in membrane cholesterol. To test this hypothesis, protocols will employ both in vivo and in vitro approaches using a variety of experimental preparations from molecular and single cell imaging studies to video-microscopy of pressurized small pulmonary arteries using a rat model of CH-induced PH. We plan to pursue the following specific aims: Specific Aim 1: Determine the mechanism by which CH decreases PASMC membrane cholesterol. Hypothesis: Elevated ROS production during CH diminishes membrane cholesterol. Specific Aim 2: Determine the mechanism by which decreased PASMC membrane cholesterol augments vasoconstrictor sensitivity following CH. Hypothesis: Decreased PASMC membrane cholesterol in response to CH unmasks EGFR-dependent pulmonary vasoconstriction through regulation of NOX2 and Rac1. The applicant will be immersed in a rich training environment in the Vascular Physiology Group at the UNM School of Medicine through a unique, multi-sponsor mentoring team that will facilitate his research training in defining novel mechanisms by which ROS alter the PASMC membrane microenvironment to affect cellular function in CH-induced PH. The proposed training plan will afford the applicant intensive training experiences in a variety of new experimental approaches, refinement of his oral and written communication skills, and professional development training that will aid him in achieving his goal as an independent, academic physician-scientist in pulmonary research.

项目概要慢性缺氧 (CH) 引起的肺动脉高压 (PH) 是发病率和死亡率的重要来源慢性阻塞性肺疾病患者的死亡率。人们普遍认为，血管收缩是 PH 的关键调节因子，尽管所涉及的机制尚不清楚。我们之前的研究已经证明，CH 后血管收缩剂敏感性的增强涉及必要的减少肺动脉平滑肌细胞（PASMC）膜胆固醇含量。我们还展示了 CH 通过改变主要依赖于钙的信号传导来增强血管收缩反应性 Ca2+ 致敏途径的机制涉及表皮生长因子受体 (EGFR) 和活性氧（ROS）。然而，CH 降低膜胆固醇和这如何揭示 EGFR 依赖性血管收缩仍有待评估。拟议的研究将调查中心假设，即血管收缩刺激与 CH 后的 EGFR 信号传导通过 ROS 依赖性减少来促进 PASMC 过度收缩膜胆固醇。为了检验这一假设，方案将采用体内和体外方法从分子和单细胞成像研究到视频显微镜的各种实验准备使用 CH 诱导的 PH 大鼠模型对小肺动脉加压。我们计划实现以下具体目标：具体目标 1：确定 CH 降低 PASMC 膜胆固醇的机制。假设：CH 期间 ROS 产生增加会减少膜胆固醇。具体目标 2：确定减少 PASMC 膜胆固醇增加的机制 CH 后的血管收缩敏感性。假设：CH 引起的 PASMC 膜胆固醇降低揭示了 EGFR 依赖性通过调节 NOX2 和 Rac1 来收缩肺血管。申请人将沉浸在新墨西哥大学血管生理学组丰富的培训环境中医学院通过一个独特的、多赞助商的指导团队，将促进他的研究培训定义ROS改变PASMC膜微环境以影响细胞的新机制在 CH 诱导的 PH 中发挥作用。拟议的培训计划将为申请人提供强化培训经验通过各种新的实验方法，提高他的口头和书面沟通技巧，以及专业发展培训将帮助他实现作为独立的学术人员的目标肺部研究的医师科学家。