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: 10029791
• 负责人: Feng Qian
• 金额: $ 58.35万
• 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/10029791
• 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; Homo; Human; Ion Channel; Ion Channel Protein; Ions; Kidney; Kidney Diseases; Knock-in Mouse; Laboratories; Life; Link; Liquid substance; Live Birth; Maps; Measures; Mediating; 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; 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.

项目摘要/摘要 我们实验室的长期目标是了解多囊蛋白蛋白的生物学功能（PC1） PC2）由常染色体显性多囊肾脏疾病（ADPKD）基因PKD1和PKD2编码 确定致病途径突变时。通过这样做，我们试图建立一个牢固的机械 了解ADPKD发病机理，可用于指导疗法的合理发展。 有效ADPKD疗法发展的巨大障碍是缺乏精确的理解 Polycystins的关键生物学功能以及其故障如何启动和驱动疾病过程。我们有 最近发现，该复合物是由PC1和PC2（PC1/PC2）形成的，两种蛋白质都衬有 通道孔，与同源PC2通道相比，用作钙渗透的离子通道 原发性进行钠和钾。这一发现表明多囊蛋白络合物获得了独特的离子 PC1组装和由此产生的PC1/PC2通道的信道特性在肾脏中起关键作用 生理学和ADPKD。在此Multi-Pi应用程序中，我们将使用新开发的功能收益（GOF） PC1/PC2通道突变体，以确定如何在分子水平调节PC1/PC2离子通道函数 通过细胞外和细胞内结构域以及Ca2+如何在该调节中起作用。我们将开发一个新的 在PC1/PC2离子通道功能中有缺陷的ADPKD鼠标模型并使用它来确定该通道 功能对于适当的肾脏发育至关重要，而该功能的灭活是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用作治疗策略，我们 将使用PC1/PC2通道功能的GOF开发另一个新的鼠标模型，并使用它来确定 它可以在模仿人ADPKD的PKD1突变小鼠中挽救该疾病。我们预计提议 研究将提供有关功能的基本分子机制和调节的新见解 PC1/PC2通道并将其通道函数定义为ADPKD的关键确定器。总体而言，该项目将 可能会更好地理解正常肾脏发育，揭示了发展的主要罪魁祸首 当PC1或PC2突变时ADPKD，并有助于构成针对PC1/PC2通道的基础 预防和治疗目的。