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: 10117237
• 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/10117237
• 关键词: 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; 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; 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.

项目摘要 /摘要 高血压是心脏病和中风的最常见危险因素。因此，重要的是要了解 高血压的发病机理，以更好地预防和治疗。罕见的孟德尔造成高血压的原因 识别以前无法识别的生理途径和网络，可以照亮新的治疗方法 疾病。孟德尔综合征家族性高钾血高血压（FHHT或假甲状腺高体性 II型）是一种由突变引起的一种单基因疾病，该疾病增加了与非赖氨酸激酶的表达 （wnks）。最近发现Cullin环连接酶（CRL）通过蛋白酶体调节血压 WNK的退化。 CRL活性的调节是由Deneddydylase COP9信号体（CSN），促进的 与复合物结合并去除泛素样蛋白NEDD8。 Cullin 3（Cul3）的突变原因 FHHT并通过体外分析显示，以增强Cul3 neddylation，增加了降解的降解 底物适配器类kelch样3（KLHL3），并降低了与CSN的结合。我们假设有障碍 与CSN的相互作用是该疾病不可或缺的。 CSN是一种精心研究的多亚基蛋白，但 这里的数据提供了第一个证据，表明肾脏中的CSN活性无序可能与高血压有关 慢性肾脏病（CKD）。申请人表征了CSN功能障碍的遗传小鼠模型， CSN的催化亚基是从肾小管细胞（KS-JAB1 - / - ）中删除的JAB1。这些老鼠 发展了不寻常的表型； WNK-Spak途径的KLHL3降低和上调 然而，类似于fhht，在整个肾单位中，JAB1删除的几种意外后果被注意到 这使观察到的表型不同于人类疾病。这包括丰度减少 几周后，NA-CL共同转运蛋白（NCC）以及进行性和自发肾脏纤维化， 模仿慢性肾脏疾病。在这里，申请人建议进一步探索这些挑衅性结果 三个具体目标。 AIM 1将继续检验以下假设，即CSN功能受损会导致FHHT 生成更忠实地模仿疾病突变的小鼠模型。在AIM 2中，体外和 体内技术将用于确定CSN是否在调节NCC中起作用。目标3意志 检查由JAB1缺失造成的肾脏损害。 NRF2积累将作为可能的研究 通过产生JAB1和NRF2双基因敲除小鼠来损害的机理。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 发展肾脏生理学研究的独立性。拟议的研究将有助于揭示机制 参与通过CUL3-KLHL3-WNK4途径调节血压的调节，这可能导致 通过靶向CRL或CSN对高血压的药物治疗。结果也将完全打开两个 新的重点领域，CSN在调节NCC退化中所扮演的作用以及扮演的潜在作用 CSN介导或加速慢性肾脏疾病的发展。