Targeting ACE2 ubiquitination for hypertension

靶向 ACE2 泛素化治疗高血压

• 批准号: 10077586
• 负责人: Catalin Filipeanu
• 金额: $ 60.05万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-26 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10077586
• 关键词: ACE2; Address; Affect; Age; Aldosterone; Angiotensin II; Blood Pressure; Bradykinin; Brain; Cardiovascular Diseases; Cardiovascular system; Cells; Cerebrovascular system; Cleaved cell; Data; Development; Dominant-Negative Mutation; Endothelial Cells; Enzymes; Family; Family member; Female; G-Protein-Coupled Receptors; Gender; Hypertension; Hypothalamic structure; Impairment; In Vitro; Individual; Inflammation; Inflammatory; Inflammatory Response; Kinins; Knockout Mice; Laboratories; Ligase; Lysine; Lysosomes; Maintenance; Mediating; Medical; Modeling; Molecular; Mus; Mutation; Neuraxis; Neurons; Peptidyl-Dipeptidase A; Pharmacology; Play; Population; Prevalence; Prevention; Proteomics; Public Health; Pulmonary Hypertension; Receptor Activation; Renin-Angiotensin System; Reporting; Resistance; Risk Factors; Role; Site; Sodium Chloride; System; Telemetry; Testing; Therapeutic; Transfection; Transgenic Model; Transgenic Organisms; Ubiquitin; Ubiquitination; United States; Validation; Vasopressins; Viral; Virus; Water; Work; aged; angiotensin I (1-7); base; conditional knockout; gene therapy; hypertension treatment; in vitro activity; in vivo; induced pluripotent stem cell; inhibitor/antagonist; male; member; mortality; mouse model; mutant; neurogenic hypertension; novel; novel strategies; optogenetics; preservation; prevent; receptor; response; salt sensitive hypertension; tool; ubiquitin isopeptidase; ubiquitin-protein ligase; vasoconstriction

With prevalence as high as 55% for individuals aged 55 and older in United States, hypertension is a major risk factor contributing to cardiovascular diseases (CVD) and global mortality, hence remaining an increasingly important medical and public health issue. The role of the renin-angiotensin system (RAS) in the maintenance of normal blood pressure (BP) and in the neuro-cardiovascular dysregulation leading to hypertension has been firmly established. Angiotensin (Ang)-II, by means of its type 1 receptor (AT1R), promotes increased sympathetic activity, salt and water reabsorption, vasoconstriction, aldosterone and vasopressin release and inflammation, all contributing to hypertension. Angiotensin Converting Enzyme type 2 (ACE2), one of the latest identified members of this system cleaves Ang-II and produces Ang-(1-7) which plays a compensatory role and opposes the RAS deleterious effects. Beyond establishing ACE2 as a critical player in the prevention of neurogenic hypertension, our group was the first to report that Ang-II mediates ACE2 ubiquitination and degradation via AT1R activation, effects that were prevented by pretreatment with a lysosomal inhibitor. Although confirmed by independent groups, the therapeutic potential of preventing ACE2 ubiquitination and degradation has not been investigated. Our pilot data, show that mutation of the C-terminus of ACE2 prevents the ubiquitination and preserves ACE2 compensatory activity while treatment with an ubiquitination-resistant ACE2 virus blunts the development of Ang-II-mediated hypertension in mice otherwise lacking ACE2. In addition, we show that targeting NEDD4, a major family of E3 ubiquitin ligases, neutralizes the deleterious effects of Ang-II on ACE2 activity, while a pilot proteomics analysis highlighted gender-specific modulation of other E3 ligases and de- ubiquitinases in the hypothalamus of mice undergoing salt-sensitive hypertension. Our preliminary data further highlight that this mechanism is not restricted to AT1R but that bradykinin B1R, activated by the inflammatory response associated to hypertension, are also involved. Thus, the hypothesis of this work is that RAS over- activation and hypertension-associated inflammation exacerbate ACE2 ubiquitination, resulting in enhanced degradation of this enzyme and a loss of its compensatory activity, ultimately reinforcing hypertension. Here, we will target ACE2 ubiquitination using novel viral approaches and unique transgenic models with an emphasis on gender- and cell-specific differences in the central nervous system and the vasculature. Validation of our hypothesis will open the door for new targeting approaches aimed at preserving ACE2 compensatory activity in hypertension and CVD.

在美国，55 岁及以上人群的患病率高达 55%，高血压是一项主要风险 导致心血管疾病（CVD）和全球死亡率的因素，因此仍然是一个日益重要的因素 重要的医疗和公共卫生问题。肾素-血管紧张素系统（RAS）在维护中的作用 正常血压（BP）和导致高血压的神经心血管失调 牢固地建立起来。血管紧张素 (Ang)-II 通过其 1 型受体 (AT1R) 促进交感神经的增强 活性、盐和水重吸收、血管收缩、醛固酮和加压素释放和炎症， 都会导致高血压。 2 型血管紧张素转换酶 (ACE2)，最新发现的酶之一 该系统的成员裂解Ang-II并产生Ang-(1-7)，Ang-(1-7)起到补偿作用并反对 RAS 的有害影响。除了将 ACE2 确立为预防神经源性疾病的关键参与者之外 高血压，我们课题组率先报道Ang-II通过介导ACE2泛素化和降解 AT1R 激活，通过溶酶体抑制剂预处理可以预防的效应。虽然证实了 独立群体中，预防 ACE2 泛素化和降解的治疗潜力尚未得到证实 调查了。我们的试验数据表明，ACE2 C 末端的突变会阻止泛素化， 保留 ACE2 补偿活性，而用抗泛素化 ACE2 病毒治疗会削弱 缺乏 ACE2 的小鼠会出现 Ang-II 介导的高血压。此外，我们还表明 靶向 NEDD4（E3 泛素连接酶的一个主要家族）可中和 Ang-II 对 ACE2 的有害影响 活动，而试点蛋白质组学分析强调了其他 E3 连接酶的性别特异性调节和去 患有盐敏感性高血压的小鼠下丘脑中的泛素酶。我们的初步数据进一步 强调这一机制不仅限于 AT1R，还包括由炎症激活的缓激肽 B1R 与高血压相关的反应也参与其中。因此，这项工作的假设是 RAS 过度 激活和高血压相关炎症会加剧 ACE2 泛素化，从而增强 这种酶的降解及其代偿活性的丧失，最终加剧高血压。在这里，我们 将使用新颖的病毒方法和独特的转基因模型来靶向 ACE2 泛素化，重点是 中枢神经系统和脉管系统的性别和细胞特异性差异。验证我们的 该假设将为新的靶向方法打开大门，旨在保留 ACE2 补偿活性 高血压和心血管疾病。