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: 10391055
• 负责人: John YJ Shyy
• 金额: $ 68.33万
• 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/10391055
• 关键词: 2019-nCoV; 5' -AMP-activated protein kinase; ACE2; Affect; Angiotensin II; Angiotensins; Animal Model; Atherosclerosis; Binding; Biology; COVID-19; Cardiovascular Diseases; Cell physiology; Cells; Cellular Metabolic Process; Cellular biology; Cues; Disease; Endothelial Cells; Endothelium; Equilibrium; 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; Pharmacology; Phosphorylation; Phosphotransferases; Physiological; Play; Post-Acute Sequelae of SARS-CoV-2 Infection; 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; extracellular; functional outcomes; in vivo; innovation; insight; interest; live cell imaging; long term consequences of COVID-19; mechanotransduction; mouse model; novel; 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)代谢是内皮功能所必需的，并不适应能量的使用 严重影响欧共体的健康。AMP激活的蛋白激酶(AMPK)是细胞能量状态的关键调节因子。 内环境平衡功能。我们的初步研究表明，能量应激导致空间上定义的AMPK 在细胞器的活性，这表明AMPK的活性在细胞内是分区的。一种新兴的 血管内皮细胞中的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，具有动脉粥样硬化保护作用。SARS冠状病毒入侵宿主 通过将病毒尖峰蛋白(S蛋白)与血管紧张素转换酶2结合，导致细胞膜血管紧张素转换酶2水平下降， 胞外可溶性ACE2增加，糖酵解增加，从而导致宿主细胞损伤。在预赛中 研究还发现，SARS-CoV-2 S蛋白使AMPK-ACE2轴失活，损伤EC 在体外和体内发挥作用。这种损害很可能构成SARS长期影响的风险因素- 冠状病毒感染或SARS-CoV-2感染的急性后遗症(PASC)。这些初步发现导致了 假设EC内环境的稳定是通过AMPK-ACE2轴的时空调节来维持的。在……里面 相比之下，S蛋白进入内皮细胞扰乱了细胞能量学，导致AMPK调控失调，进而导致 血管紧张素转换酶2低磷酸化，这在新冠肺炎相关的EC功能障碍和PASC中起关键作用。 提出用来检验这一新假说的三个具体目标如下：目标1.调查 生理[例如，脉动切应力]下内皮细胞AMPK的时空调节， 药理学条件(如二甲双胍)和病理生理条件(如S蛋白)；目的2.破译 生理、药理和病理生理刺激调节AMPK的机制-- 目的3.探讨AMPK-ACE2轴受损在S蛋白加速损伤中的作用 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， 并在活体动物模型中确定AMPK-ACE2轴在内皮健康和疾病中的作用。 这些发现将导致对PASC的病理生理学缺乏深入的了解，这将继续 是SARS-CoV-2感染的长期后果。