AMPK Regulation of ACE2 in Endothelial Health and Disease

AMPK 在内皮健康和疾病中对 ACE2 的调节

• 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
• 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' -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

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.

项目摘要 血管内皮细胞（EC）代谢是功能性内皮细胞和不适应的能量使用所必需的 严重影响健康。AMP活化蛋白激酶（AMPK）是细胞能量状态的关键调节因子， 自我平衡功能我们的初步研究表明，能量应激导致空间上确定的AMPK 在细胞器中的AMPK活性，这表明AMPK活性在细胞中是区室化的。一个新兴 血管内皮中AMPK的底物是血管紧张素转换酶2（ACE 2），我们发现 AMPK-ACE 2轴增强EC功能并具有动脉粥样硬化保护作用。SARS-CoV病毒侵入宿主 通过将病毒刺突蛋白（S蛋白）与ACE 2结合，导致细胞膜ACE 2水平降低， 增加的细胞外可溶性ACE 2和增加的糖酵解，从而导致宿主细胞损伤。初步 研究中，我们还发现SARS-CoV-2S蛋白使AMPK-ACE 2轴失活， 在体外和体内发挥作用。这种损害可能构成SARS长期影响的风险因素- CoV-2感染或SARS-CoV-2感染后急性后遗症（PASC）。这些初步调查结果导致 假设EC稳态通过AMPK-ACE 2轴的时空调节来维持。在 相反，S蛋白的进入破坏了EC中的细胞能量学，导致AMPK失调， ACE 2低磷酸化，这对COVID-19相关的EC功能障碍和PASC至关重要。 为检验这一新假设而提出的三个具体目标如下：目标1。探讨 在生理条件下EC中AMPK的时空调节[例如，脉动剪切应力（PS）]， 药理学的（例如，二甲双胍），和病理生理学（例如，S蛋白）条件;目的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ACE 2轴; Aim 3。研究AMPK-ACE 2轴受损在S蛋白加速的 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， 和体内动物模型，以确定AMPK-ACE 2轴在内皮健康和疾病中的作用。 这些发现将导致对PASC病理生理学的其他见解缺失，这将继续 是SARS-CoV-2感染的长期后果。