Structural and functional integrity and microenvironment of RPE cells

RPE 细胞的结构和功能完整性以及微环境

基本信息

批准号：
8415828
负责人：
CHING-HWA SUNG
金额：
$ 46.59万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-08-01 至 2016-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8415828
关键词：
3-Dimensional Active Biological Transport Address Adhesions Adverse effects Age Age related macular degeneration Aging Animal Model Animals Apical Area Atrophic Biochemical Biological Models Blindness Blood capillaries Bruch&apos s basal membrane structure Cell Communication Cell Culture Techniques Cell Line Cell membrane Cell physiology Cells Choroid Communication Complex Deposition Development Diffusion Disease Disease Progression Doxycycline Early Diagnosis Elderly Endothelial Cells Epithelial Epithelial Cells Etiology Exocytosis Future Gene Expression Guanosine Triphosphate Phosphohydrolases High Prevalence Homeostasis In Situ Integral Membrane Protein Knockout Mice Lead Link Maintenance Measures Mediating Membrane Membrane Protein Traffic Methods Modeling Mus Names Neonatal Pathology Pathway interactions Pattern Permeability Phenotype Photoreceptors Physiologic pulse Physiological Plant Roots Plasmids Play Primary Lesion Protein Secretion Proteins Reporter Retina Retinal Retinal Detachment Rodent Rodent Model Role Site Structure Structure of retinal pigment epithelium Surface Testing Time Tissues Transfection Transgenic Mice Vascular Endothelial Growth Factors Vision apical membrane base capillary cell type cellular microvillus extracellular human CLIC4 protein in vivo innovation insight macular edema mouse model mutant novel protein transport relating to nervous system senescence small hairpin RNA therapy design trafficking young adult

项目摘要

DESCRIPTION (provided by applicant): The retinal pigment epithelium (RPE) is a multifunctional and indispensable component of the vertebrate retina. The convoluted apical and basal plasma membranes of RPE cells provide a large surface area that allows rapid material exchange between these cells and their local microenvironment. The RPE is considered to be the primary lesion site of age-related macular degeneration (AMD), a disease that has pathology spanning the entire photoreceptor-RPE-choriocapillaris complex. The root cause of the initial damage to the RPE remains unclear. Because the RPE cells have an active transport role, we propose that a decline in membrane trafficking is sufficient to cause numerous adverse effects on membrane specialization, protein trafficking, and/or secretion. These effects could be conveyed to neighboring cells through direct cell-cell interaction and/or diffusion. Chloride intracellular channel 4 (CLIC4), an apical RPE protein, has an important role in vesicular exocytosis in other epithelial cell types. To study the physiological relevance of CLIC4, we have developed two novel rodent models in which CLIC4 can be selectively suppressed from RPE cells in situ (i.e., in vivo transfection, and conditional knockout mice). In both of these model systems, young adults manifest several cell autonomous and non-cell autonomous features that not only mirror each other, but also mimic the hallmarks of AMD. To build upon these findings, we will conduct a comprehensive characterization of these animals to better model the disease progression of AMD (Aim 1). Furthermore, we will directly test our model that the dysregulated vesicular trafficking function of CLIC4 is what causes the microvillar dysmorphogenesis and retinal detachment in the mutant animals (Aim 2). Finally, we will test the hypothesis that CLIC4 is important for the secretion of molecules produced by RPE cells (Aim 3). Imbalanced secretion may lead to atrophy in the adjacent tissues. Several innovative techniques, cell cultures, and state-of-the-art animal models will be used in combination to address these inter-related questions. These studies will enrich our fundamental understanding of the RPE and ultimately lead to better diagnosis of early AMD and rational design of treatments.

描述（申请人提供）：视网膜色素上皮（RPE）是脊椎动物视网膜的多功能且不可或缺的组成部分。 RPE 细胞的回旋顶端和基底质膜提供了很大的表面积，允许这些细胞与其局部微环境之间进行快速的物质交换。 RPE 被认为是年龄相关性黄斑变性 (AMD) 的主要病变部位，AMD 是一种病理学跨越整个光感受器-RPE-脉络膜毛细血管复合体的疾病。 RPE 最初受损的根本原因仍不清楚。由于 RPE 细胞具有主动运输作用，我们认为膜运输的下降足以对膜特化、蛋白质运输和/或分泌造成许多不利影响。这些效应可以通过直接的细胞间相互作用和/或扩散传递到邻近的细胞。氯离子胞内通道 4 (CLIC4) 是一种顶端 RPE 蛋白，在其他上皮细胞类型的囊泡胞吐作用中具有重要作用。为了研究 CLIC4 的生理相关性，我们开发了两种新型啮齿动物模型，其中 CLIC4 可以被原位 RPE 细胞选择性抑制（即体内转染和条件敲除小鼠）。在这两个模型系统中，年轻人表现出多种细胞自主和非细胞自主特征，这些特征不仅相互镜像，而且模仿 AMD 的特征。为了以这些发现为基础，我们将对这些动物进行全面的表征，以更好地模拟 AMD 的疾病进展（目标 1）。此外，我们将直接测试我们的模型，即 CLIC4 失调的囊泡运输功能是导致突变动物中微绒毛畸形发生和视网膜脱离的原因（目标 2）。最后，我们将检验 CLIC4 对于 RPE 细胞产生的分子的分泌很重要的假设（目标 3）。分泌不平衡可能导致邻近组织萎缩。将结合使用几种创新技术、细胞培养和最先进的动物模型来解决这些相互关联的问题。这些研究将丰富我们对 RPE 的基本了解，并最终更好地诊断早期 AMD 并合理设计治疗方案。