AMPK Regulation of ACE2 in Endothelial Health and Disease

AMPK 在内皮健康和疾病中对 ACE2 的调节

基本信息

批准号：
10568995
负责人：
John YJ Shyy
金额：
$ 67.31万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
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：
2022-02-15 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10568995
关键词：
2019-nCoV 5&apos -AMP-activated protein kinase ACE2 Acceleration Affect Angiotensin II Angiotensins Animal Model Arterial Lines Atherosclerosis Binding Biology COVID-19 Cardiovascular Diseases Cell physiology Cells Cellular Metabolic Process Cellular biology Cues Disease Endothelial Cells Endothelium Functional disorder Glycolysis Goals Health Heart Diseases Homeostasis Impairment In Vitro Inflammation Invaded Lead Link Long-Term Effects Membrane Metformin Molecular Mutant Strains Mice Organelles Oxidation-Reduction Phosphorylation Phosphotransferases Physiological Play Post-Acute Sequelae of SARS-CoV-2 Infection Proliferating Protein Kinase Proteins Regulation Renin-Angiotensin-Aldosterone System Reporter Research Risk Factors Role SARS coronavirus SARS-CoV-2 infection SARS-CoV-2 spike protein Signal Transduction Stimulus Stress Testing Up-Regulation Vascular Endothelial Cell Vascular Endothelium Vasodilation Viral Virus angiogenesis arm atheroprotective cardiovascular health cell injury cell type energy balance extracellular functional outcomes in vivo innovation insight interest live cell imaging long term consequences of COVID-19 mechanotransduction mouse model novel pharmacologic shear stress spatiotemporal tool upstream kinase

项目摘要

Project Summary Vascular endothelial cell (EC) metabolism is essential for functional endothelium, and maladapted energy use severely affects EC health. AMP-activated protein kinase (AMPK) is a key regulator of cellular energy status and homeostatic function. Our preliminary studies showed that energy stress results in spatially defined AMPK activity at cellular organelles, which indicates that AMPK activity is compartmentalized in the cell. An emerging AMPK substrate in the vascular endothelium is angiotensin-converting enzyme 2 (ACE2), and we have found that the AMPK–ACE2 axis enhances EC function and is atheroprotective. SARS-CoV viruses invade the host cells by binding the viral spike protein (S protein) to ACE2, which leads to decreased membrane ACE2 levels, increased extracellular soluble ACE2, and increased glycolysis, thus resulting in host cell damage. In preliminary studies, we have also found that the SARS-CoV-2 S protein deactivates the AMPK–ACE2 axis and impairs EC function in vitro and in vivo. This impairment is likely to constitute a risk factor for the long-term effects of SARS- CoV-2 infection or post-acute sequelae of SARS-CoV-2 infection (PASC). These preliminary findings lead to the hypothesis that EC homeostasis is maintained via the spatiotemporal regulation of the AMPK–ACE2 axis. In contrast, S protein entry disrupts cellular energetics in ECs, leading to dysregulated AMPK and the ensuing ACE2 hypo-phosphorylation, which critically contributes to the COVID-19–associated EC dysfunction and PASC. The three specific aims proposed to test this novel hypothesis are as follows: Aim 1. To investigate the spatiotemporal regulation of AMPK in ECs under physiological [e.g., pulsatile shear stress (PS)], pharmacological (e.g., metformin), and pathophysiological (e.g., S protein) conditions; Aim 2. To decipher the mechanisms by which physiological, pharmacological, and pathophysiological stimuli modulate the AMPK– ACE2 axis in ECs; Aim 3. To investigate the role of impaired AMPK–ACE2 axis in S protein-accelerated atherosclerosis in the context of PASC. In the proposed research, we will use live cell imaging, in vitro EC biology, and in vivo animal models to determine the role of the AMPK–ACE2 axis in endothelial health and disease. These findings will result in otherwise missing insights into the pathophysiology of PASC, which will continue to be a long-term consequence of SARS-CoV-2 infection.

项目概要血管内皮细胞 (EC) 代谢对于功能性内皮细胞和不适应的能量使用至关重要严重影响 EC 健康。AMP 激活蛋白激酶 (AMPK) 是细胞能量状态的关键调节因子。我们的初步研究表明，能量应激会导致空间定义的 AMPK。细胞器中的活性，这表明 AMPK 活性在细胞中是划分的。血管内皮中的AMPK底物是血管紧张素转换酶2（ACE2），我们发现 AMPK-ACE2 轴增强 EC 功能并对 SARS-CoV 病毒入侵宿主具有保护作用。通过将病毒刺突蛋白（S 蛋白）与 ACE2 结合，导致细胞膜 ACE2 水平降低，细胞外可溶性ACE2增加，糖酵解增加，从而初步导致宿主细胞损伤。研究中，我们还发现 SARS-CoV-2 S 蛋白会使 AMPK-ACE2 轴失活并损害 EC 这种损伤可能构成 SARS 长期影响的危险因素。这些初步发现导致了 CoV-2 感染或 SARS-CoV-2 感染的急性后遗症（PASC）。假设 EC 稳态是通过 AMPK-ACE2 轴的时空调节来维持的。相比之下，S 蛋白的进入会破坏 EC 中的细胞能量，导致 AMPK 失调以及随之而来的 ACE2 低磷酸化，对 COVID-19 相关的 EC 功能障碍和 PASC 起着至关重要的作用。为检验这一新假设而提出的三个具体目标如下：目标 1. 调查生理条件下 EC 中 AMPK 的时空调节 [例如，脉动剪切应力 (PS)]，药理学（例如二甲双胍）和病理生理学（例如 S 蛋白）条件；目标 2. 破译生理学、药理学和病理生理学刺激调节 AMPK 的机制 EC 中的 ACE2 轴；目标 3. 研究受损的 AMPK-ACE2 轴在 S 蛋白加速中的作用 PASC 背景下的动脉粥样硬化在拟议的研究中，我们将使用活细胞成像、体外 EC 生物学、以及体内动物模型，以确定 AMPK-ACE2 轴在内皮健康和疾病中的作用。这些发现将导致人们对 PASC 病理生理学的认识缺失，而 PASC 的病理生理学将继续是 SARS-CoV-2 感染的长期后果。