Determining the role of afadin in tubular development and maintenance in the kidney

确定阿法丁在肾小管发育和维护中的作用

• 批准号: 10675599
• 负责人: Isabel A Alejandra Lopez-Garcia
• 金额: $ 5.27万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10675599
• 关键词: 3-Dimensional; Adherens Junction; Apical; Architecture; Automobile Driving; Binding; Cells; Central cord canal structure; Chronic Kidney Failure; Cytoskeleton; Data; Defect; Development; Dialysis procedure; Drug Implants; Epithelium; Evolution; F-Actin; Filtration; Future; Goals; Grant; Image; Implant; In Vitro; Intercellular Junctions; Kidney; Kidney Transplantation; Knock-out; Knockout Mice; Knowledge; Label; Lead; Liquid substance; Maintenance; Mediating; Modeling; Molecular; Morphogenesis; Mus; Nephrons; Organoids; Patients; Process; Proteins; Regulation; Renal Replacement Therapy; Renal function; Renal tubule structure; Research; Role; Scaffolding Protein; Signal Transduction; Structure; System; Testing; Therapeutic; Tissues; Tube; Tubular formation; Visualization; Work; afadin; burden of illness; clinically relevant; design; follow-up; global health; improved; in vitro Model; in vivo; in vivo Model; innovation; mouse model; nephrogenesis; novel therapeutic intervention; novel therapeutics; renal epithelium; replacement tissue; tissue regeneration; trafficking

Project Summary/Abstract Chronic kidney disease (CKD) poses an important global disease burden with limited therapeutic options. Novel therapeutic strategies to improve renal function include implanting patient-derived kidney organoids onto the native kidneys. However, an inability to connect organoid tubules with the host’s kidney tubules presents a major problem that remains to be solved. Currently, the molecular mechanisms driving interconnection between tubules and their lumens are poorly understood. Filling this gap in knowledge is critical to advance work with implantable renal replacement tissues. Our previous studies have shown that mice lacking afadin, a cytoskeletal scaffolding protein, from renal epithelial tubules, have a defect in lumen continuity. The goal of this project is to determine how lumen connection/fusion occurs within tubules and determine the molecular mechanism by which afadin promotes lumen fusion and maintenance in renal epithelia. We hypothesize that lumen fusion requires cellular rearrangements and that these are facilitated by afadin-mediated regulation of cell-cell contacts. Here, I propose to use well-defined in vitro and in vivo models to: (1) Determine the mechanism of lumen fusion after the onset of de novo lumenogenesis, (2) Identify the afadin domains required for lumen fusion, and (3) Determine the role of afadin in lumen maintenance in vitro and in an in vivo inducible knockout mouse model. The knowledge obtained from these studies will enable a better design of epithelial structures that have the ability to form and maintain a continuous lumen for future use as implantable therapeutics.

项目概要/摘要 慢性肾病 (CKD) 是一种重要的全球疾病负担，但治疗选择有限。 改善肾功能的治疗策略包括将源自患者的肾脏类器官植入到 然而，无法将类器官肾小管与宿主的肾小管连接起来是一个主要问题。 目前，驱动小管之间互连的分子机制仍有待解决。 对它们的流明知之甚少，填补这一知识空白对于推进植入式工作至关重要。 我们之前的研究表明，小鼠缺乏阿法丁（一种细胞骨架支架）。 来自肾上皮小管的蛋白质在管腔连续性方面存在缺陷。该项目的目标是确定。 管腔连接/融合如何在肾小管内发生，并确定 afadin 的分子机制 促进肾上皮细胞的管腔融合和维持我们发现管腔融合需要细胞。 重排，并且这些是通过 afadin 介导的细胞间接触调节来促进的。 使用明确的体外和体内模型来：（1）确定发病后管腔融合的机制 de novo lumenogenesis，（2）识别腔融合所需的 afadin 结构域，以及（3）确定作用 阿法丁在体外和体内诱导敲除小鼠模型中管腔维持的知识。 从这些研究中获得的结果将有助于更好地设计具有形成和形成能力的上皮结构。 保持连续的管腔以供将来用作植入疗法。