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

项目总结/摘要 慢性肾脏病（CKD）是一种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源自患者的肾类器官植入到肾组织上。 原生肾脏然而，不能将类器官小管与宿主的肾小管连接， 这个问题还有待解决。目前，驱动小管之间相互连接的分子机制 而它们的内腔却知之甚少。填补这一知识空白对于推进植入式药物的研究至关重要。 肾脏替代组织我们之前的研究表明，缺乏afadin（一种细胞骨架支架）的小鼠 来自肾上皮小管的蛋白质在管腔连续性方面有缺陷。这个项目的目标是确定 管腔连接/融合如何在小管内发生，并确定afadin 促进肾上皮细胞的管腔融合和维持。我们假设管腔融合需要细胞 在某些实施方案中，细胞与细胞之间的相互作用是通过细胞-细胞接触的afadin介导的调节来促进的。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）鉴定管腔融合所需的afadin结构域，（3）确定 在体外和体内可诱导敲除小鼠模型中，知识 从这些研究中获得的细胞将能够更好地设计上皮结构， 保持连续的管腔以供将来用作可植入治疗剂。

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.