Regulation of vascular leakage in age related macular degeneration

年龄相关性黄斑变性中血管渗漏的调节

基本信息

批准号：
8385057
负责人：
MEREDITH GREGORY-KSANDER
金额：
$ 30.02万
依托单位：
SCHEPENS EYE RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8385057
关键词：
Address Age related macular degeneration Age-Years Agonist American Angiogenic Factor Animal Model Apoptosis Apoptotic Atrophic Binding Binding Proteins Blindness Blood Blood Vessels Cell surface Cells Choroidal Neovascularization Cleaved cell Data Development Doxycycline Endothelial Cells Extravasation Eye Failure Fluorescein Angiography Gene Targeting Goals Growth Immune Induction of Apoptosis Inflammatory Inflammatory Infiltrate Integral Membrane Protein Interleukin-1 Isolectin Knock-in Mouse Laboratories Lasers Lead Lesion Liquid substance Maintenance Mediating Membrane Metalloproteases Modeling Mouse Strains Mus Mutate Optical Coherence Tomography Pathogenesis Pathway interactions Patients Phenotype Play Population Protein Isoforms Proteins Recombinants Regulation Reporting Retina Retinal Role Site Staining method Stains Structure of retinal pigment epithelium System TNF gene Therapeutic Time Tissues Transgenic Mice Tumor Necrosis Factor Ligand Superfamily Member 6 Vascular Endothelial Cell Vascular Endothelial Growth Factors Vision Wild Type Mouse Work bevacizumab cell type cytokine cytotoxic extracellular in vivo intravitreal injection macrophage mouse model neovascular new growth new therapeutic target novel overexpression photoreceptor degeneration prevent protein function success

Address Age related macular degeneration Age-Years Agonist American Angiogenic Factor Animal Model Apoptosis Apoptotic Atrophic Binding Binding Proteins Blindness Blood Blood Vessels Cell surface Cells Choroidal Neovascularization Cleaved cell Data Development Doxycycline Endothelial Cells Extravasation Eye Failure Fluorescein Angiography Gene Targeting Goals Growth Immune Induction of Apoptosis Inflammatory Inflammatory Infiltrate Integral Membrane Protein Interleukin-1 Isolectin Knock-in Mouse Laboratories Lasers Lead Lesion Liquid substance Maintenance Mediating Membrane Metalloproteases Modeling Mouse Strains Mus Mutate Optical Coherence Tomography Pathogenesis Pathway interactions Patients Phenotype Play Population Protein Isoforms Proteins Recombinants Regulation Reporting Retina Retinal Role Site Staining method Stains Structure of retinal pigment epithelium System TNF gene Therapeutic Time Tissues Transgenic Mice Tumor Necrosis Factor Ligand Superfamily Member 6 Vascular Endothelial Cell Vascular Endothelial Growth Factors Vision Wild Type Mouse Work bevacizumab cell type cytokine cytotoxic extracellular in vivo intravitreal injection macrophage mouse model neovascular new growth new therapeutic target novel overexpression photoreceptor degeneration prevent protein function success

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项目摘要

DESCRIPTION (provided by applicant): Age-related macular degeneration (AMD) is a leading cause of blindness in Americans 60 years of age and older. There are two forms of AMD: Dry or atrophic and Wet or exudative. However, the Wet form of AMD, which is characterized by choroidal neovascularization (CNV), is associated with more severe vision loss caused primarily by the leakage of fluid and blood from the abnormal vessels. Anti-VEGF therapy has emerged as the treatment of choice for patients with wet AMD. However, some patients never respond to anti-VEGF therapy, while still others stop responding after initial success, highlighting the fact that the pathobiology of wet AMD development and progression is still unknown. FasL is an important protein in maintaining immune privilege in the eye, where it is thought to induce apoptosis of infiltrating inflammatory cells. In fact, several reports indicate that FasL expressed on retinal pigment epithelial cells inhibits choroidal neovascularization by triggering apoptosis o vascular endothelial cells. However, FasL exists as both a membrane bound and soluble protein with potentially opposing functions and it is unclear how the different forms of FasL contribute to the development of CNV. We recently developed a unique knock-in mouse strain in which the FasL metalloproteinase cleavage sites were mutated to prevent cleavage of the membrane-bound protein. In these ¿CS mice, FasL is expressed by the physiologically relevant cell types, but these cells are unable to cleave FasL and therefore, can only express mFasL (termed ¿CS mice). For the first time, these mice have allowed us to study the in vivo function of membrane FasL in the absence of soluble FasL. To better understand the function of the different FasL isoforms in the development of choroidal neovascularization, we used a laser-induced murine model of CNV. Our preliminary data indicate that, in ¿CS mice, either the increased expression of mFasL (or the loss of sFasL) prevents vascular leakage following laser induced-CNV. Spectral domain optical coherence tomography (SD-OCT) and Isolectin-B staining demonstrated no difference in the size of the neovascular lesion between ¿CS mice and wild-type mice. However, fluorescein angiography revealed a significant decrease in vascular leakage in ¿CS mice as compared to wild-type mice. These results indicate that membrane FasL does not prevent the ingrowth of choroidal vessels, as previously documented, but rather plays a critical function in preventing vascular leakage, the primary cause of vision loss in patients with wet AMD. We hypothesize that membrane FasL inhibits vascular leakage in laser-induced CNV through Fas mediated pathways triggered in infiltrating macrophages and/or RPE cells. The goals of this proposal are as follows: (Aim 1) Determine whether the decreased vascular leakage in ¿CS mice is due to increased mFasL and/or a lack of sFasL and whether the decreased leakage coincides with a shift in the M1/M2 phenotype of infiltrating macrophages (Aim 2) Establish whether infiltrating macrophages and/or RPE are the critical Fas+ targets through which mFasL mediates inhibition of vascular leakage. PUBLIC HEALTH RELEVANCE: Age Related Macular Degeneration (AMD) is the primary cause of blindness in patients 60 years of age and older. The growth of abnormal blood vessels into the retina is the main cause of severe vision loss, in part due to vessel leakage of blood and fluid that causes damage to the surrounding retinal tissue. We have identified a novel protein that prevents vascular leakage and the goal of this study is to (i) determine whether the administration of this protein can be used to prevent vascular leakage in a mouse model, and (ii) elucidate the mechanism by which this protein functions.

描述（由适用提供）：与年龄相关的黄斑变性（AMD）是60岁及以上的美国人失明的主要原因。 AMD有两种形式：干燥或萎缩，潮湿或先进。然而，以脉络膜新生血管形成（CNV）为特征的AMD的湿形形式与更严重的视力丧失有关，主要是由于异常血管中流体和血液泄漏而引起的。抗VEGF疗法已成为湿AMD患者的首选治疗方法。但是，有些患者从未对抗VEGF疗法做出反应，而另一些患者在最初成功后停止反应，强调了湿AMD发育和进展的病理学仍然未知的事实。 FASL是维持眼睛中免疫特权的重要蛋白质，在那里人们认为它会诱导浸润性炎症细胞的凋亡。实际上，有几份报告表明FASL表示在视网膜色素上的上皮细胞上，通过触发凋亡o血管内皮细胞抑制脉络膜新血管化。但是，FASL既是具有可能相反功能的膜结合和固体蛋白的存在，尚不清楚不同形式的FASL如何有助于 CNV的发展。我们最近开发了一种独特的敲入小鼠菌株，其中将FASL金属蛋白酶裂解位点突变以防止膜结合蛋白的裂解。在这些CS小鼠中，FASL由物理相关的细胞类型表达，但是这些细胞无法清除FASL，因此只能表达MFASL（称为CS小鼠）。这些小鼠首次使我们能够在没有固体FASL的情况下研究膜FASL的体内功能。为了更好地理解不同FASL同工型在脉络膜新生血管形成发展中的功能，我们使用了激光诱导的CNV鼠模型。我们的初步数据表明，在CS小鼠中，MFASL的表达增加（或SFASL丢失）可以防止激光诱导CNV后血管泄漏。光谱结构域光学相干断层扫描（SD-OCT）和分离蛋白-B染色表明，CS小鼠和野生型小鼠之间的新生血管病变的大小没有差异。然而，与野生型小鼠相比，荧光素血管造影显示CS小鼠的血管泄漏显着下降。这些结果表明，如前所述，膜FASL并不能阻止脉络膜视频的侵蚀，而是在防止血管泄漏方面起着关键功能，这是湿AMD患者视力丧失的主要原因。我们假设膜FASL抑制激光诱导的CNV中的血管渗漏，这通过FAS介导的途径在浸润巨噬细胞和/或RPE细胞中触发。该提案的目标如下：（目标1）确定€cs小鼠中血管泄漏减少是由于MFASL增加和/或缺乏SFASL引起的，以及减少的泄漏是否与M1/M2表型的变化是否同时发生，是否与Mocrophages的M1/M2表型相一致（AIM 2）确定了Indriltrating Mocltrating Mocltrating Mocltrating Mocltrating Mocltrating Moctrating Mope+ RPE是否是/或RPE。介导抑制血管泄漏。公共卫生相关性：与年龄相关的黄斑变性（AMD）是60岁及以上患者失明的主要原因。异常血管进入视网膜的生长是严重视力丧失的主要原因，部分原因是血管泄漏和造成周围视网膜组织损害的流体。我们已经确定了一种防止血管泄漏的新型蛋白质，这项研究的目的是（i）确定该蛋白的给药是否可用于防止小鼠模型中的血管泄漏，以及（ii）阐明该蛋白质功能的机制。