Role of the COP9 Signalosome (CSN) in kidney disease and hypertension

COP9 信号体 (CSN) 在肾脏疾病和高血压中的作用

• 批准号: 10583478
• 负责人: Ryan J Cornelius
• 金额: $ 12.97万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-02 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10583478
• 关键词: Acceleration; American; Area; Award; Binding; Blood Pressure; COPS5 gene; Catalytic Domain; Cells; Chronic Kidney Failure; Complex; Cullin Proteins; Cultured Cells; DNA Sequence Alteration; Data; Development; Disease; Distal; Distal convoluted renal tubule structure; Environment; Excision; Experimental Designs; Failure; Familial disease; Functional disorder; Future; Genetic; Goals; Heart Diseases; Homeostasis; Human; Hypertension; Impairment; In Vitro; Journals; Kidney; Kidney Diseases; Kidney Failure; Knockout Mice; Lead; Ligase; Lysine; Mediating; Medical; Mendelian disorder; Mentors; Modeling; Molecular; Mus; Mutation; Nephrology; Nephrons; Pathogenesis; Pathway interactions; Pharmacologic Substance; Phenotype; Phosphotransferases; Physiological; Physiology; Play; Positioning Attribute; Potassium; Prevention; Protein Subunits; Proteins; Public Health; Publishing; Regulation; Renal tubule structure; Research; Research Personnel; Risk Factors; Role; Site; Societies; Sodium Chloride; Stroke; Syndrome; Techniques; Testing; Type II Pseudohypoaldosteronism; Ubiquitin; Ubiquitin Like Proteins; Up-Regulation; Work; blood pressure regulation; cost efficient; cullin-3; disease phenotype; familial hyperkalemic hypertension; human disease; hyperkalemia; hypertension treatment; in vivo; in vivo evaluation; interest; kidney fibrosis; mouse model; multicatalytic endopeptidase complex; mutant; novel therapeutics; prevent; protein degradation; renal damage; skills; success; symporter; thiazide; ubiquitin ligase; ubiquitin-protein ligase

Project Summary / Abstract Hypertension is the most common risk factor for heart disease and stroke; therefore, it is important to understand the pathogenesis of hypertension for better prevention and treatment. Rare Mendelian causes of hypertension identify previously unrecognized physiological pathways and networks, which can illuminate new treatments for disease. The Mendelian syndrome Familial Hyperkalemic Hypertension (FHHt, or pseudohypoaldosteronism type II) is a monogenic disease resulting from mutations which increase expression of with-no-lysine kinases (WNKs). Cullin-RING ligases (CRLs) were recently discovered to regulate blood pressure via proteasomal degradation of WNKs. Regulation of CRL activity is facilitated by the deneddylase, COP9 Signalosome (CSN), which binds to the complex and removes the ubiquitin-like protein, NEDD8. A mutation in cullin 3 (CUL3) causes FHHt and was shown, by in vitro analysis, to have enhanced CUL3 neddylation, increased degradation of the substrate adaptor kelch-like 3 (KLHL3), and decreased binding to the CSN. We hypothesized that the impaired interaction with the CSN was integral to the disease. The CSN is a well-studied multi-subunit protein, but the data here provides the first evidence that disordered CSN activity in the kidney may relate to hypertension and chronic kidney disease (CKD). The applicant has characterized a genetic mouse model of CSN dysfunction, in which the catalytic subunit of the CSN, Jab1 is deleted from kidney-tubule cells (KS-Jab1-/-). These mice developed an unusual phenotype; there was decreased KLHL3 and upregulation of the WNK-SPAK pathway akin to FHHt, however, several unexpected consequences of Jab1 deletion throughout the nephron were noted that made the observed phenotype differ from the human disease. This included a decrease in the abundance of the Na-Cl co-transporter (NCC) after several weeks, and progressive and spontaneous kidney fibrosis, mimicking chronic kidney disease. Here, the applicant proposes to further explore these provocative results in three specific aims. Aim 1 will continue to test the hypothesis that impaired CSN function causes FHHt by generating mouse models that more faithfully mimic the disease mutation. In Aim 2, a combination of in vitro and in vivo techniques will be used to determine whether the CSN plays a role in modulating NCC directly. Aim 3 will examine the kidney damage caused by deletion of Jab1. Nrf2 accumulation will be investigated as a possible mechanism for the damage by generating Jab1 and Nrf2 double knockout mice. In addition to successful completion of these aims the mentoring and scientific environment make the applicant an ideal candidate to develop independence in renal physiology research. The proposed research will help reveal the mechanisms involved in regulation of blood pressure through the CUL3-KLHL3-WNK4 pathway which could lead to pharmaceutical treatment of hypertension by targeting CRLs, or the CSN. The results will also open two entirely new areas of focus, the role that the CSN plays in regulating NCC degradation and the potential role played by the CSN in mediating or accelerating the development of chronic kidney disease.

项目概要/摘要 高血压是心脏病和中风最常见的危险因素；因此，了解这一点很重要 了解高血压的发病机制，以便更好地预防和治疗。高血压的罕见孟德尔病因 识别以前未被识别的生理途径和网络，这可以阐明新的治疗方法 疾病。孟德尔综合征家族性高钾血症性高血压（FHHt，或假性醛固酮增多症 II 型）是一种单基因疾病，由增加无赖氨酸激酶表达的突变引起 （WNK）。最近发现 Cullin-RING 连接酶 (CRL) 可通过蛋白酶体调节血压 WNK 的降解。 CRL 活性的调节由 deneddylase、COP9 Signalosome (CSN) 促进， 它与复合物结合并去除泛素样蛋白 NEDD8。 cullin 3 (CUL3) 突变导致 FHHt 并通过体外分析显示，增强了 CUL3 neddylation，增加了 底物接头 kelch-like 3 (KLHL3)，并减少与 CSN 的结合。我们假设受损者 与 CSN 的相互作用是该疾病不可或缺的一部分。 CSN 是一种经过充分研究的多亚基蛋白，但 这里的数据提供了第一个证据，证明肾脏中 CSN 活性紊乱可能与高血压和 慢性肾脏病（CKD）。申请人已经表征了 CSN 功能障碍的遗传小鼠模型， 其中 CSN 的催化亚基 Jab1 从肾小管细胞中删除 (KS-Jab1-/-)。这些老鼠 形成不寻常的表型； KLHL3 减少，WNK-SPAK 通路上调 然而，与 FHHt 类似，我们注意到整个肾单位中 Jab1 缺失的一些意想不到的后果 这使得观察到的表型与人类疾病不同。这包括丰度的减少 几周后 Na-Cl 协同转运蛋白 (NCC) 的变化，以及进行性和自发性肾纤维化， 模仿慢性肾脏疾病。在此，申请人建议进一步探索这些具有挑战性的结果 三个具体目标。目标 1 将继续检验 CSN 功能受损导致 FHHt 的假设： 生成更忠实地模仿疾病突变的小鼠模型。在目标 2 中，体外和 体内技术将用于确定 CSN 是否在直接调节 NCC 中发挥作用。目标3将 检查 Jab1 缺失引起的肾脏损伤。将调查 Nrf2 积累是否可能 通过产生 Jab1 和 Nrf2 双敲除小鼠来研究损伤机制。除了成功 完成这些目标的指导和科学环境使申请人成为理想的候选人 培养肾脏生理学研究的独立性。拟议的研究将有助于揭示其机制 通过 CUL3-KLHL3-WNK4 通路参与血压调节，这可能导致 通过针对 CRL 或 CSN 进行高血压的药物治疗。结果也将完全打开两个 新的关注领域、CSN 在调节 NCC 降解中所发挥的作用以及 CSN 介导或加速慢性肾病的发展。