The Role of EPB41L5 in Regulation of Cilia Function

EPB41L5 在纤毛功能调节中的作用

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

概括发育缺陷是美国乃至全世界发病率和死亡率的重要原因。虽然在开始了解有助于实现这一目标的基础生物学方面已经取得了相当大的进展对于这些缺陷，还有很多仍知之甚少。我们研究细胞重新排列的形态发生细胞骨架组织响应周围环境的信号。这个过程中的缺陷在胚胎发生过程中会导致出生缺陷和发育问题。我们的长期目标是更全面了解形态发生：细胞如何响应来自环境和环境的信号重塑发育中胚胎的细胞骨架，其中细胞骨架的动态重排是形态发生的主要驱动力。这对于增进我们对先天性和细胞骨架重排是缺陷的根本原因的产后疾病。这个提议研究旨在探索红细胞膜蛋白条带 4.1 like 5 (Epb41l5) 在睫状体功能的调节。纤毛是从顶端表面延伸出来的毛发状延伸物，用于接收信号。 Epb41l5 是介导胞浆蛋白与质膜蛋白结合的支架蛋白。 Epb41l5 调节许多需要肌动蛋白细胞骨架重塑的细胞过程，顶皮质的特殊肌动蛋白。虽然之前的研究报道了 epb41l5 缺失小鼠的纤毛功能障碍胚胎中，这些研究得出的结论是纤毛缺陷继发于其他 Epb41l5 的缺陷诸如顶端-基底极性形成等功能。然而，我们的初步数据表明 Epb41l5 可以对睫状体功能有直接的调节作用。我们建立了斑马鱼 epb41l5 突变体的新等位基因显示正常的顶端基底极性形成，但显示与纤毛功能障碍相关的表型。我们将肾囊肿素 5 (NHPH5) 鉴定为一种新型 Epb41l5 相互作用蛋白。 NPHP5 突变为最初在肾结核患者中发现，NPHP5 已被证明可以调节纤毛功能。我们的假设是：Epb41l5 通过抑制 NPHP5 在纤毛上的定位来调节纤毛功能。我们将测试使用哺乳动物细胞培养系统和斑马鱼胚胎的假设，这是一种已建立的脊椎动物遗传可在所有发育阶段进行实验操作和成像的模型。完成这项研究将进一步了解睫状体功能的调节机制，这对于制定更有效的治疗策略来对抗纤毛病。我们认为这项研究成本很高有效并将产生为未来 R01 奠定基础的数据。