The Role of EPB41L5 in Regulation of Cilia Function

EPB41L5 在纤毛功能调节中的作用

• 批准号: 10053505
• 负责人: Miho Matsuda
• 金额: $ 8.48万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-11-07 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10053505
• 关键词: Actins; Affect; Alleles; Apical; Binding; Biology; C-terminal; Cell Culture System; Cell Culture Techniques; Cell Shape; Cell membrane; Cell physiology; Cell surface; Cells; Cilia; Congenital Abnormality; Cystic kidney; Cytoplasmic Protein; Cytoskeletal Modeling; Cytoskeleton; Data; Defect; Development; Disease; Embryo; Embryonic Development; Environment; Erythrocyte Membrane; Exhibits; Failure; Functional disorder; Future; Genes; Genetic; Genetic Models; Goals; Hair; Handedness; Hepatobiliary; Human; Image; Investigation; Knockout Mice; Lead; Leber' s amaurosis; Mammalian Cell; Mediating; Membrane Proteins; Morbidity - disease rate; Morphogenesis; Mus; Mutation; Names; Nephronophthisis; Patients; Phenotype; Process; Proteins; Regulation; Reporting; Research; Retina; Role; Scaffolding Protein; Secondary to; Signal Transduction; Source; Surface; Syndrome; Testing; Treatment Efficacy; Zebrafish; base; cell motility; ciliopathy; combat; cost effective; driving force; effective therapy; human disease; mortality; mutant; new therapeutic target; novel; novel therapeutic intervention; overexpression; postnatal; response; synergism

Summary Developmental defects are substantial contributors to morbidity and mortality in the US as well as worldwide. While considerable progress has been made to begin to understand the underlying biology that contributes to these defects, much remains poorly understood. We study morphogenesis in which cells rearrange cytoskeletal organization in response to signals from its surrounding environments. Defects in this process during embryogenesis cause birth defects and developmental problems. Our long-term goal is a more comprehensive understanding of morphogenesis: how a cell responds to signals from its environment and remodels the cytoskeleton in developing embryos where dynamic rearrangement of the cytoskeleton is the major driving force of morphogenesis. This is essential to advance our understanding of congenital and postnatal disorders in which cytoskeletal rearrangement is the underlying source of the defect. This proposed research is aimed at exploring a novel role of Erythrocyte membrane protein band 4.1 like 5 (Epb41l5) in the regulation of ciliary function. Cilia are hair like extensions from the apical surface to receive signals. Epb41l5 is a scaffold protein that mediates association of cytosolic proteins with proteins at the plasma membrane. Epb41l5 regulates a number of cellular processes that require remodeling of the actin cytoskeleton, in particular actin at the apical cortex. While previous studies reported cilia dysfunction in epb41l5 null mouse embryos, these studies concluded that the defects in cilia were secondary to defects of other Epb41l5 functions such as apical-basal polarity formation. Our preliminary data, however, suggests that Epb41l5 could have a direct role in regulating ciliary function. We established novel alleles of zebrafish epb41l5 mutants which showed normal apicobasal polarity formation but showed phenotypes associated with cilia dysfunction. We identified Nephrocystin 5 (NHPH5) as a novel Epb41l5 interacting protein. NPHP5 mutations were originally identified in patients with Nephronophthisis and NPHP5 has shown to regulate cilia function. Our hypothesis is that: Epb41l5 regulates ciliary function by inhibiting NPHP5 localization at cilia. We will test the hypothesis using mammalian cell culture system and zebrafish embryos, an established vertebrate genetic model that is accessible for experimental manipulation and imaging at all developmental stages. Completion of this study will lead to further understanding of regulatory mechanisms of ciliary function, which is necessary for developing more effective therapeutic strategies to combat ciliopathies. We believe that this study is highly cost effective and will produce data that lays the basis for a future R01.

总结 发育缺陷是美国和世界范围内发病率和死亡率的重要因素。 虽然已经取得了相当大的进展，开始了解潜在的生物学，有助于 这些缺陷，很多仍然知之甚少。我们研究细胞重新排列的形态发生 细胞骨架组织响应来自其周围环境的信号。此过程中的缺陷 在胚胎发育过程中会导致出生缺陷和发育问题。我们的长期目标是 形态发生的全面理解：细胞如何响应来自环境的信号， 在发育中的胚胎中重塑细胞骨架，其中细胞骨架的动态重排是 形态发生的主要驱动力。这对于促进我们对先天性和 细胞骨架重排是缺陷的潜在来源的产后疾病。这一拟议 本研究旨在探索红细胞膜蛋白带4.1样5（Epb 4115）在红细胞凋亡中的新作用。 调节纤毛功能。纤毛是从顶端表面延伸的毛发状延伸物，用于接收信号。Epb 41 l5是 一种介导细胞溶质蛋白与质膜蛋白结合的支架蛋白。 Epb 41 l5调节许多需要肌动蛋白细胞骨架重塑的细胞过程， 特别是顶端皮层的肌动蛋白。虽然先前的研究报道了epb 41 l5敲除小鼠的纤毛功能障碍， 胚胎，这些研究得出结论，纤毛的缺陷是继发于其他Epb 41 l5缺陷 如顶基极性形成的功能。然而，我们的初步数据表明，Epb 4115可能 在调节纤毛功能中具有直接作用。我们建立了斑马鱼epb 41 l5突变体的新等位基因， 其显示正常的顶基极性形成，但显示与纤毛功能障碍相关的表型。 我们鉴定了一种新的Epb 4115相互作用蛋白--肾囊蛋白5（NHPH 5）。NPHP 5突变是 最初在患有肾病的患者中鉴定，并且NPHP 5已显示出调节纤毛功能。我们 假设：Epb 4115通过抑制NPHP 5在纤毛上的定位来调节纤毛功能。我们将测试 假设使用哺乳动物细胞培养系统和斑马鱼胚胎，一个建立的脊椎动物遗传 该模型可用于所有发育阶段的实验操作和成像。完成 这项研究将导致进一步了解纤毛功能的调节机制，这是必要的， 开发更有效的治疗策略来对抗纤毛病。我们认为，这项研究的成本很高， 有效，并将产生为未来R 01奠定基础的数据。