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

项目摘要/摘要 慢性肾脏疾病(CKD)是一个重要的全球疾病负担，治疗选择有限。小说 改善肾功能的治疗策略包括将患者衍生的肾脏器官移植到 天生的肾脏。然而，无法将器官小管与宿主的肾小管连接起来是一种主要的 这是有待解决的问题。目前，驱动小管之间相互连接的分子机制 人们对它们的管腔知之甚少。填补这一知识空白对于推进可植入材料的工作至关重要 肾脏替代组织。我们之前的研究表明，缺乏细胞骨架支架Fafadin的小鼠 来自肾上皮小管的蛋白质在管腔连续性方面存在缺陷。这个项目的目标是确定 管腔连接/融合在小管内如何发生，并决定afadin的分子机制 促进肾上皮细胞管腔融合和维持。我们假设管腔融合需要细胞 这些重排是由afadin介导的细胞-细胞接触的调节来促进的。在这里，我建议 使用明确的体外和体内模型：(1)确定发病后管腔融合的机制 (2)确定管腔融合所需的afadin结构域，以及(3)确定其作用 在体外和体内可诱导的基因敲除小鼠模型中非他丁在管腔维持中的作用。《知识》 从这些研究中获得的结果将使更好地设计具有形成和 保持连续的管腔，以供将来作为植入性治疗使用。

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.