Role of VEGFR2 trafficking in pathological angiogenesis in age related macular degeneration

VEGFR2 运输在年龄相关性黄斑变性病理性血管生成中的作用

基本信息

批准号：
10376188
负责人：
Kaori Horiguchi Yamada
金额：
$ 38.78万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
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：
2019-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10376188
关键词：
Adherens Junction Age related macular degeneration Blindness Blood Vessels Cell membrane Cells Choroidal Neovascularization Clinical Cues Data Defect Disease Disease Progression Endothelial Cells Extravasation Eye Filopodia Foundations Genetic Goals Growth Image Inflammation KDR gene Kinesin Knockout Mice Lasers Lead Ligands Light Mediating Membrane Modeling Molecular Molecular Motors Motor Mus Pathogenesis Pathologic Pathologic Neovascularization Pathology Pathway interactions Patients Peptides Permeability Pharmacology Plus End of the Microtubule Protein Family Regulation Retina Role Signal Transduction Site Stimulus Testing VEGFA gene Vascular Endothelial Growth Factors Work angiogenesis base cadherin 5 cell type drug development genetic approach inhibitor insight live cell imaging mouse model neovascularization novel novel strategies novel therapeutic intervention older patient prevent receptor response retinal damage targeted treatment tool trafficking

项目摘要

ABSTRACT Wet age-related macular degeneration (AMD) is a leading cause of vision loss in old patients. In wet AMD, excessive vascular endothelial growth factor (VEGF) causes abnormal angiogenesis and vascular leakage, which in turn damages the retina. The overall goal of this proposal is to determine the mechanism how excessive VEGF induces transition from quiescent blood vessels to pathological leaky angiogenesis in wet AMD by studying the fundamental roles of VEGFR2 trafficking. Angiogenesis is restrained in quiescent healthy endothelial cells (ECs), where VEGFR2 trafficking is limited by the interaction with VE-cadherin at adherens junctions (AJs). In contrast, marked VEGFR2 trafficking is evident in angiogenic ECs, where VEGFR2 translocates to filopodia tips that extend towards the VEGF ligand. We provided the first evidence that VEGFR2 is directly transported by the kinesin-3 family protein, KIF13B, a microtubules plus-end motor, to filopodia of sprouting ECs. Based on our finding, we will test our central hypothesis that KIF13B mediates VEGFR2 trafficking away from AJs to induce AJ disassembly and vascular leakage, and the directional trafficking of VEGFR2 to filopodia induces pathological angiogenesis in response to excessive VEGF in wet AMD. Our Specific Aims will test the following hypotheses; 1) KIF13B-mediated VEGFR2 trafficking from AJs breaks the critical interaction between VEGFR2 and VE-cadherin involved in stabilizing AJs, thus induces AJ disassembly and vascular leakage. 2) VEGF signaling induces KIF13B-mediated directional trafficking of VEGFR2 to filopodia extending toward VEGF, and the trafficking is required for sprouting angiogenesis. 3) KIF13B-mediated VEGFR2 trafficking is pathogenesis in wet AMD, thus the inhibition of the trafficking is a promising strategy for the therapy of wet AMD. To rigorously test these hypotheses, our lab has generated powerful tools (genetic mouse models and peptide inhibitors) that have led to conceptual advances. EC specific KIF13B knockout mice display a selective angiogenic defect in the pathological setting. A small peptide inhibitor disrupting the KIF13B/VEGFR2 interaction, termed KAI, inhibited choroidal neovascularization (CNV) in wet AMD model in mice. Using these powerful tools, we will examine the roles of KIF13B in VEGF-induced permeability of ocular blood vessels, live imaging of directional VEGFR2 trafficking in choroidal sprouting ex vivo, and pathology of wet AMD, characterized by abnormal angiogenesis, vascular leakage, and inflammation, using laser-induced CNV model. If successful, our proposed studies provide the novel concept of angiogenesis regulation by targeting VEGFR2 trafficking.

抽象的湿性年龄相关性黄斑变性（AMD）是老年患者视力丧失的主要原因。在湿 AMD 中，过量的血管内皮生长因子（VEGF）导致血管生成异常和血管渗漏，这反过来又会损害视网膜。该提案的总体目标是确定该机制如何过度 VEGF 通过以下方式诱导湿性 AMD 中从静止血管向病理性渗漏血管生成的转变研究 VEGFR2 贩运的基本作用。静态健康血管生成受到抑制内皮细胞 (EC)，其中 VEGFR2 运输受到粘附层 VE-钙粘蛋白相互作用的限制路口 (AJ)。相比之下，显着的 VEGFR2 转运在血管生成 EC 中很明显，其中 VEGFR2 易位至丝状伪足尖端，向 VEGF 配体延伸。我们提供了第一个证据表明 VEGFR2 由驱动蛋白 3 家族蛋白 KIF13B（一种微管正端马达）直接转运至丝状伪足发芽的 EC。根据我们的发现，我们将检验我们的中心假设：KIF13B 介导 VEGFR2 远离 AJ 的贩运导致 AJ 解体和血管渗漏，以及定向贩运 VEGFR2 与丝状伪足的结合可诱导病理性血管生成，以响应湿性 AMD 中过量的 VEGF。我们的具体目标将检验以下假设； 1) KIF13B 介导的 AJ 中的 VEGFR2 运输打破了 VEGFR2 和 VE-钙粘蛋白之间的关键相互作用参与稳定 AJ，从而诱导 AJ 解体和血管渗漏。 2) VEGF信号传导诱导KIF13B介导的VEGFR2向丝状伪足的定向运输向 VEGF 延伸，并且运输是萌发血管生成所必需的。 3) KIF13B介导的VEGFR2 转运是湿性 AMD 的发病机制，因此抑制转运是一种有前途的治疗策略湿性AMD。为了严格检验这些假设，我们的实验室开发了强大的工具（基因小鼠模型和肽抑制剂）带来了概念上的进步。 EC 特异性 KIF13B 敲除小鼠表现出病理环境中的选择性血管生成缺陷。一种破坏 KIF13B/VEGFR2 的小肽抑制剂这种相互作用（称为 KAI）可抑制小鼠湿性 AMD 模型中的脉络膜新生血管（CNV）。使用这些作为强大的工具，我们将研究 KIF13B 在 VEGF 诱导的眼部血管通透性中的作用，活体离体脉络膜萌芽中定向 VEGFR2 运输的成像，以及湿性 AMD 的病理学，使用激光诱导的 CNV 模型，以异常血管生成、血管渗漏和炎症为特征。如果成功，我们提出的研究提供了通过靶向 VEGFR2 来调节血管生成的新概念贩运。