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: 10543403
• 负责人: Isabel A Alejandra Lopez-Garcia
• 金额: $ 5.18万
• 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/10543403
• 关键词: 3-Dimensional; Adherens Junction; Apical; Architecture; Automobile Driving; Binding; Cells; Central cord canal structure; Chronic Kidney Failure; Data; Defect; Development; Dialysis procedure; Epithelial; Evolution; F-Actin; Filtration; Future; Goals; Grant; Heel; 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; Work; afadin; base; 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.

项目摘要/摘要 慢性肾脏疾病（CKD）具有重要的全球疾病伯恩，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植入 本地肾脏。但是，无法将类正弦管与宿主的肾小管连接起来。 尚待解决的问题。目前，分子机制驱动管子之间的互连 他们的流明知之甚少。在知识中填补这一差距对于推进植入式工作至关重要 肾脏替代组织。我们以前的研究表明，缺乏afadin的小鼠是细胞骨架脚手架 来自肾上皮管的蛋白质在管腔连续性中缺陷。该项目的目的是确定 管腔连接/融合如何发生在管中，并确定afadin的分子机制 促进肾上皮中的管腔融合和维护。我们假设管腔融合需要细胞 重排和这些是通过Afadin介导的细胞触点调节制备的。我建议在这里 使用明确定义的体外和体内模型以：（1）确定发作后管腔融合的机理 （2）识别腔蛋白融合所需的afadin域，（3）确定作用 在体外和体内诱导敲除小鼠模型中，腔内维持中的afadin。知识 从这些研究中获得的将使具有形成能力的上皮结构进行更好的设计 保持连续的管腔供将来用作植入疗法。