Ion Channel Function and Regulation of the Polycystin-1/2 Complex in Kidney Physiology and Polycystic Kidney Disease

多囊蛋白-1/2复合物在肾脏生理学和多囊肾病中的离子通道功能和调节

• 批准号: 10665001
• 负责人: Feng Qian
• 金额: $ 53.82万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10665001
• 关键词: Affect; Amino Acids; Architecture; Autosomal Dominant Polycystic Kidney; Binding; Biochemical; Biological Assay; Biological Process; C-terminal; Calcium; Cations; Coiled-Coil Domain; Complex; Coupling; Cryoelectron Microscopy; Cyst; Cystic kidney; Development; Disease; EF Hand Motifs; Electrophysiology (science); Ensure; G-Protein-Coupled Receptors; Goals; Human; Ion Channel; Ion Channel Protein; Ions; Kidney; Kidney Diseases; Knock-in Mouse; Laboratories; Life; Link; Liquid substance; Live Birth; Maps; Measures; Mediating; Membrane; Methods; Molecular; Morphogenesis; Morphology; Mutant Strains Mice; Mutate; Mutation; N-terminal; Nephrons; PKD1 gene; PKD2 gene; PKD2 protein; Pathogenesis; Pathogenicity; Pathway interactions; Permeability; Physiology; Play; Polycystic Kidney Diseases; Potassium; Preventive; Process; Property; Proprotein Convertase 1; Proprotein Convertase 2; Proteins; Proteolysis; Publishing; Regulation; Renal function; Role; Signal Pathway; Signal Transduction; Site; Sodium; Structure; System; TRP channel; Tail; Testing; Therapeutic; Uncertainty; Xenopus oocyte; extracellular; gain of function; in vivo; insight; loss of function; mouse model; mutant; nephrogenesis; new therapeutic target; novel; polycystic kidney disease 1 protein; preservation; prevent; protein complex; receptor; stoichiometry; success; targeted treatment; therapeutic target; therapy development

Project Summary/Abstract The long-term goals of our laboratories are to understand the biological functions of polycystin proteins (PC1 and PC2) encoded by autosomal dominant polycystic kidney disease (ADPKD) genes PKD1 and PKD2, and to determine the pathogenic pathways when they are mutated. By so doing, we seek to establish a firm mechanistic understanding of the ADPKD pathogenesis, which can be used to guide the rational development of therapies. A great obstacle to the development of effective ADPKD therapies has been the lack of a precise understanding of polycystins’ key biological function and how its malfunction initiates and drives the disease process. We have most recently discovered that the complex formed by PC1 and PC2 (PC1/PC2), with both of proteins lining the channel pore, functions as a calcium-permeable ion channel, in contrast to the homomeric PC2 channel which primarily conducts sodium and potassium. This finding indicates that the polycystin complex gains unique ion channel properties from the assembly of PC1 and the resulting PC1/PC2 channel plays a key role in kidney physiology and ADPKD. In this multi-PI application, we will use our newly developed gain-of-function (GOF) PC1/PC2 channel mutant to determine how the PC1/PC2 ion channel function is regulated at a molecular level by extracellular and intracellular domains and how Ca2+ plays a role in this regulation. We will develop a new ADPKD mouse model that is defective in PC1/PC2 ion channel function and use it to determine that the channel function is essential for proper kidney development and inactivation of this function is the real culprit for ADPKD. To determine whether enhancing the PC1/PC2 ion channel function can be used as a therapeutic strategy, we will develop another new mouse model with the GOF of PC1/PC2 channel function and use it to determine that it can rescue the disease in a PKD1 mutant mouse that mimics human ADPKD. We anticipate that the proposed studies will provide new insights into the fundamental molecular mechanism of function and regulation of the PC1/PC2 channel and define its channel function as the key determinant of ADPKD. Overall, the project will likely lead to a better understanding of normal kidney development, reveal the primary culprit for developing ADPKD when PC1 or PC2 are mutated, and help form the basis for targeting the PC1/PC2 channel for preventative and therapeutic purposes.

项目概要/摘要 我们实验室的长期目标是了解多囊蛋白（PC1 和 PC2) 由常染色体显性多囊肾病 (ADPKD) 基因 PKD1 和 PKD2 编码，并且 确定突变时的致病途径。通过这样做，我们寻求建立牢固的机制 了解 ADPKD 发病机制，可用于指导合理开发治疗方法。 开发有效的 ADPKD 疗法的一个巨大障碍是缺乏准确的理解 多囊蛋白的关键生物学功能及其故障如何启动和驱动疾病过程。我们有 最近发现，由 PC1 和 PC2 (PC1/PC2) 形成的复合物，两种蛋白质均排列在 通道孔，起到钙渗透离子通道的作用，与同聚 PC2 通道相反， 主要传导钠和钾。这一发现表明多囊蛋白复合物获得了独特的离子 PC1 组装的通道特性以及由此产生的 PC1/PC2 通道在肾脏中发挥着关键作用 生理学和 ADPKD。在此多 PI 应用中，我们将使用我们新开发的功能增益 (GOF) PC1/PC2 通道突变体，用于确定 PC1/PC2 离子通道功能如何在分子水平上受到调节 细胞外和细胞内结构域的作用以及 Ca2+ 如何在这种调节中发挥作用。我们将开发一个新的 PC1/PC2离子通道功能缺陷的ADPKD小鼠模型，并用它来确定该通道 功能对于肾脏的正常发育至关重要，而该功能的失活是 ADPKD 的真正罪魁祸首。 为了确定增强 PC1/PC2 离子通道功能是否可以作为一种治疗策略，我们 将开发另一种具有PC1/PC2通道功能GOF的新鼠标模型，并用它来确定 它可以挽救模仿人类 ADPKD 的 PKD1 突变小鼠的疾病。我们预计拟议的 研究将为了解功能和调节的基本分子机制提供新的见解。 PC1/PC2通道并定义其通道功能作为ADPKD的关键决定因素。总体而言，该项目将 可能有助于更好地了解正常肾脏发育，揭示发育的罪魁祸首 ADPKD 当 PC1 或 PC2 发生突变时，有助于形成针对 PC1/PC2 通道的基础 预防和治疗目的。

项目成果

The heteromeric PC-1/PC-2 polycystin complex is activated by the PC-1 N-terminus.

• DOI: 10.7554/elife.60684
• 发表时间: 2020-11-09
• 期刊: eLife
• 影响因子: 7.7
• 作者: Ha K;Nobuhara M;Wang Q;Walker RV;Qian F;Schartner C;Cao E;Delling M
• 通讯作者: Delling M

The roles of two extracellular loops in proton sensing and permeation in human Otop1 proton channel.

• DOI: 10.1038/s42003-022-04085-2
• 发表时间: 2022-10-20
• 期刊: COMMUNICATIONS BIOLOGY
• 影响因子: 5.9
• 作者: Li, Bin;Wang, Yan;Castro, Alexis;Ng, Courtney;Wang, Zhifei;Chaudhry, Haroon;Agbaje, Zainab;Ulloa, Gabriella A.;Yu, Yong
• 通讯作者: Yu, Yong

The diverse effects of pathogenic point mutations on ion channel activity of a gain-of-function polycystin-2.

• DOI: 10.1016/j.jbc.2023.104674
• 发表时间: 2023-05
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Wang, Yan;Wang, Zhifei;Pavel, Mahmud Arif;Ng, Courtney;Kashyap, Parul;Li, Bin;Morais, Tiago D. C.;Ulloa, Gabriella A.;Yu, Yong
• 通讯作者: Yu, Yong

The ion channel TRPM7 regulates zinc-depletion-induced MDMX degradation.

• DOI: 10.1016/j.jbc.2021.101292
• 发表时间: 2021-11
• 期刊: The Journal of biological chemistry
• 作者: Wang H;Li B;Asha K;Pangilinan RL;Thuraisamy A;Chopra H;Rokudai S;Yu Y;Prives CL;Zhu Y
• 通讯作者: Zhu Y

Fibrocystin/Polyductin releases a C-terminal fragment that translocates into mitochondria and suppresses cystogenesis.

• DOI: 10.1038/s41467-023-42196-4
• 发表时间: 2023-10-16
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Walker, Rebecca, V;Yao, Qin;Xu, Hangxue;Maranto, Anthony;Swaney, Kristen F.;Ramachandran, Sreekumar;Li, Rong;Cassina, Laura;Polster, Brian M.;Outeda, Patricia;Boletta, Alessandra;Watnick, Terry;Qian, Feng
• 通讯作者: Qian, Feng