Targeting Mitochondrial Cyclophilin D in Vascular Oxidative Stress and Hypertension

靶向线粒体亲环蛋白 D 治疗血管氧化应激和高血压

• 批准号: 10449107
• 负责人: Sergey Dikalov
• 金额: $ 42.1万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10449107
• 关键词: Acetylation; Acetyltransferase; Adult; Angiotensin II; Animal Model; Antihypertensive Agents; Antioxidants; Arteries; Attenuated; Blood Pressure; Blood Vessels; CRISPR/Cas technology; Cells; Clinical Data; DOCA; Data; Deacetylase; Deacetylation; Development; Disease; Drug Targeting; Endothelium; Essential Hypertension; Female; Genetic; Health; Heart failure; Human; Hypertension; Inflammation; Inner mitochondrial membrane; Interdisciplinary Study; Knock-out; Knockout Mice; Lipids; Measures; Mediating; Mitochondria; Mitochondrial Matrix; Modeling; Molecular; Mus; Mutant Strains Mice; Myocardial Infarction; Organ; Oxidative Stress; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Population; Positioning Attribute; Regulation; Relaxation; Residual state; Resistance; Risk Factors; Role; Smooth Muscle; Societies; Sodium Chloride; Stress; Stroke; Superoxides; Testing; Therapeutic; Tissues; Transgenic Mice; Transgenic Organisms; Vascular Diseases; Vasodilation; Wild Type Mouse; Work; blood pressure reduction; cyclophilin D; cytokine; endothelial dysfunction; human subject; human tissue; hypertension treatment; hypertensive; improved; inhibitor; male; mitochondrial dysfunction; mitochondrial permeability transition pore; mouse model; multiple drug use; novel; oxidation; prevent; targeted agent; therapeutic evaluation

Project Summary Hypertension is a major health problem in Western Societies and a risk factor for stroke, myocardial infarction, and heart failure and therapies specifically targeted at mitochondria represent promising strategies to reduce target- organ-damage. Mitochondrial permeability transition pore (mPTP) plays a key role in mitochondrial dysfunction and target-organ-damage in hypertension. We discovered that depletion or inhibition of Cyclophilin D (CypD), a regulatory subunit of mPTP opening, improves vascular function and attenuates hypertension. Meanwhile, the tissue specific role of CypD and molecular mechanisms of CypD activation are not known. In the proposed studies, we will take this work forward by defining the role of vascular CypD and therapeutic potential to target CypD in endothelial dysfunction and hypertension. Our studies in human subjects with essential hypertension showed CypD hyperacetylation and implicate CypD activation by K166 acetylation due to imbalance between GCN5L1 acetyltransfe- rase and reduced Sirt3 deacetylase activity. Sirt3 is inactivated by highly reactive mitochondrial lipid dicarbonyls, isolevuglandins (isoLG), while isoLG scavenging prevents CypD acetylation and reduces hypertension. We propose targeting mitochondrial CypD to inhibit oxidative stress, improve vascular functions and reduce hypertension. The overall objective of this proposal is to define specific mechanisms of CypD mediated vascular oxidative stress and test the therapeutic potential of targeting CypD in hypertension. We will pursue the following aims: AIM 1. To test the hypothesis that cell specific CypD depletion in endothelial and smooth muscle reduces vascular oxidative stress, protects vascular relaxation and attenuates hypertension. In this aim we will examine the protective role of CypD depletion in inducible endothelial specific CypD knockout (EcCypDKO) and smooth muscle specific CypD knockout (SmcCypDKO) mice using AngII and DOCA-salt models of hypertension. AIM 2. To test the hypothesis that CypD-K166 acetylation contributes to vascular dysfunction and hypertension. We will define the pathophysiological significance of CypD-K166 acetylation using new deacetylation mimic CypD-K166R mutant mice, new endothelial specific GCN5L1 knockout mice (EcGCN5L1KO), and endothelial specific Sirt3 knockout mice (EcSirt3KO). All mice are available in our lab. AIM 3. To test the hypothesis that CypD inhibition and blocking CypD hyperacetylation after onset hypertension improve vascular function. We will test if CypD blockers improve vascular function and reduce blood pressure in hypertensive mice. We will study CypD acetylation in resistance arteries isolated from human subjects with essential hypertension and test if CypD blockers improve human endothelial function. We are in an ideal position to perform these interdisciplinary studies. We developed new CypD transgenic mouse models and mitochondria-targeted treatments. We have access to human vascular tissue and unique expertise in oxidative stress, human vascular studies and hypertension. Our data strongly support this novel pathway in vascular dysfunction, and this work has the potential to make a major impact on the development of new treatments.

项目摘要 高血压是西方社会的主要健康问题，也是中风、心肌梗死和脑梗死的危险因素。 心力衰竭和特异性靶向线粒体治疗代表了减少靶向线粒体的有希望的策略， 器官损伤线粒体通透性转换孔（mitochondrialpermeability transition pore，mPTP）在线粒体功能障碍中起关键作用 和靶器官损害。我们发现，亲环素D（CypD），一种 mPTP开放调节亚基，改善血管功能，减轻高血压。同时 CypD的组织特异性作用和CypD活化的分子机制尚不清楚。在拟议的研究中， 我们将通过定义血管CypD的作用和靶向CypD的治疗潜力来推进这项工作， 内皮功能障碍和高血压。我们对原发性高血压患者的研究表明CypD 由于GCN 5L 1乙酰转移与K166乙酰化之间不平衡， 降低Sirt 3脱乙酰酶活性。Sirt 3被高度反应性的线粒体脂质二羰基化合物灭活， 异左旋木素（isoLG），而isoLG清除阻止CypD乙酰化并降低高血压。我们提出 靶向线粒体CypD，抑制氧化应激，改善血管功能，降低高血压。的 该提案的总体目标是确定CypD介导的血管氧化应激的具体机制 并测试靶向CypD在高血压中的治疗潜力。我们将努力实现以下目标： AIM 1.为了检验内皮和平滑肌中细胞特异性CypD耗竭减少血管内皮细胞和平滑肌细胞中的细胞特异性CypD的假设， 氧化应激，保护血管舒张和减轻高血压。为此，我们将研究 CypD耗竭在诱导型内皮特异性CypD敲除（EcCypDKO）和平滑中的保护作用 肌肉特异性CypD敲除（SmcCypDKO）小鼠，使用AngII和DOCA-盐高血压模型。 AIM 2.为了检验CypD-K166乙酰化导致血管功能障碍和高血压的假设。我们 将使用新的去乙酰化模拟物确定CypD-K166乙酰化的病理生理学意义 CypD-K166 R突变小鼠、新内皮特异性GCN 5L 1敲除小鼠（EcGCN 5L 1 KO）和内皮细胞 特异性Sirt 3敲除小鼠（EcSirt 3 KO）。所有的老鼠都在我们的实验室里。 AIM 3.为了验证高血压发作后CypD抑制和阻断CypD高乙酰化的假设， 改善血管功能。我们将测试CypD阻滞剂是否能改善血管功能， 高血压小鼠的血压。我们将研究CypD乙酰化在人的阻力动脉中的作用 受试者患有原发性高血压，并测试CypD阻断剂是否改善人内皮功能。 我们处于一个理想的位置来进行这些跨学科的研究。我们研制出新型CypD转基因小鼠 模型和靶向治疗。我们有机会接触到人体血管组织和独特的专业知识 氧化应激、人体血管研究和高血压。我们的数据强烈支持这种新的途径， 血管功能障碍，这项工作有可能对新治疗方法的发展产生重大影响。