RETINA: REVERSED POLARITY AND MORPHOGENESIS OF RPE

视网膜：RPE 的极性反转和形态发生

• 批准号: 7269795
• 负责人: Enrique J Rodriguez-Boulan
• 金额: $ 56.45万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-03-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7269795
• 关键词: Accounting; Adaptor Signaling Protein; Address; Adenovirus Infections; Adhesions; Adult; Amino Acid Receptors; Amino Acid Transporter; Apical; Basolateral Sorting Signal; Cell Line; Cell membrane; Cells; Chloride Ion; Chlorides; Cultured Cells; Cyst; Data; Disease; Epithelial Cells; Epithelium; Golgi Apparatus; In Situ; Integrins; Ion Transport; Laboratories; Lactate Transporter; Low Density Lipoprotein Receptor; MDCK cell; Membrane Proteins; Microtubules; Modeling; Molecular; Molecular Chaperones; Morphogenesis; Neural Cell Adhesion Molecules; Neutral Amino Acid Transport Systems; Nutrient; Pathway interactions; Phenotype; Physiology; Predisposition; Protein Sortings; Proteins; Protons; RNA Interference; Research; Research Personnel; Retina; Retinal; Role; Route; Signal Transduction; Simple Epithelium; Sorting - Cell Movement; Structure of retinal pigment epithelium; Tars; Transferrin Receptor; Tyrosine; Virus Receptors; adenovirus receptor; base; basolateral membrane; cdc42 GTP-Binding Protein; gene therapy; improved; intracellular protein transport; kidney cell; novel; pH Homeostasis; programs; protein kinase D; protein transport; prototype; receptor; receptor recycling; research study; symporter; syntaxin 1; syntaxin 4; trafficking; uptake; vesicular stomatitis virus G protein

DESCRIPTION (provided by applicant): This proposal addresses the fundamental mechanisms involved in the polarized trafficking of plasma membrane proteins in Retinal Pigment Epithelium (RPE). Previous research from our laboratory has revealed dramatic differences in the targeting of several apical and basolateral plasma membrane proteins between RPE and other simple epithelia, represented by the prototype kidney cell line MDCK. Basolateral proteins such as the neural cell adhesion molecule (NCAM) and the chaperone for lactate transporters CD147 (EMMPRIN) have a strikingly reversed apical polarity in adult RPE. Experiments in SA1 will study the role of different basolateral sorting signals in polarized transport between Golgi and plasma membrane in RPE and study the role of CD147 in the polarized sorting of neutral amino acid transporters. Experiments in SA2 will explore the role of basolateral sorting adaptors in RPE. We will explore the RPE trafficking phenotype arising from the absence of AP1B, an adaptor found in most other epithelial cells. This phenotype includes alterations in the trafficking of Transferrin Receptors, compared to AP1B+ epithelial cells, and a dramatically increased susceptibility to adenovirus infection due to altered trafficking in virus receptors. Other experiments will characterize the basolateral sorting role of other adaptors present in RPE (APIA, AP3, AP4, GGA1-3), using an RNAi approach, and search for novel adaptors/proteins that may account for the reversed polarity of NCAM and CD147. Experiments in SA3 will address the mechanisms involved in polarized vesicular delivery in RPE. This aim builds on significant technical and conceptual advances in our understanding of the vesicular delivery machinery of MDCK cells. We will characterize the role of svntaxins 1 and 4, microtubules, cdc42 and IQGAP1 in protein sorting by RPE cells. We will explore the hypothesis that some components of the basolateral vesicular trafficking machinery in MDCK cells may operate in reverse in RPE cells. The data obtained will provide fundamental information on how RPE cells organize their polarized trafficking routes. This information is essential to understand RPE's roles in the physiology of the retina and in blinding diseases, and may help in devising improved gene therapy strategies for their treatment.

描述（由申请人提供）：该提案解决了视网膜色素上皮（RPE）中质膜蛋白两极分化的基本机制。我们实验室的先前研究表明，在RPE和其他简单上皮中，以原型肾细胞系MDCK表示，RPE和其他简单上皮菌之间的几种顶端和基底外侧质膜蛋白的靶向差异很大。基底外侧蛋白，例如神经细胞粘附分子（NCAM）和乳酸转运蛋白CD147（Emmprin）的伴侣蛋白在成人RPE中具有显着逆转的根尖极性。 SA1中的实验将研究RPE中高尔基体和质膜之间极化转运中不同基底外侧分类信号的作用，并研究CD147在中性氨基酸转运蛋白的极化排序中的作用。 SA2中的实验将探讨RPE中基底外侧分类适配器的作用。我们将探索由于不存在AP1B而引起的RPE运输表型，AP1B是在大多数其他上皮细胞中发现的适配器。与AP1B+上皮细胞相比，该表型包括转移蛋白受体的运输的改变，以及由于病毒受体的运输改变而引起的腺病毒感染的易感性大大提高。其他实验将使用RNAi方法（APIA，AP3，AP4，GGA1-3）中存在的其他适配器的基底外侧排序作用，并使用RNAi方法，并搜索新的适配器/蛋白质，这些适配器/蛋白可能占NCAM和CD147的极性。 SA3中的实验将解决RPE偏振囊泡递送所涉及的机制。这个目标是基于我们对MDCK细胞囊泡传递机制的重要技术和概念进步的基础。我们将表征SVNTAXINS 1和4，微管，CDC42和IQGAP1在RPE细胞中分类中的作用。我们将探讨以下假设：MDCK细胞中基底外侧囊泡运输机制的某些组成部分可能会在RPE细胞中反向运行。获得的数据将提供有关RPE细胞如何组织其两极分化途径的基本信息。这些信息对于了解RPE在视网膜生理和盲目疾病中的作用至关重要，并且可能有助于制定改进的基因疗法治疗策略。