Delineating the role of TIMP3 in macular degeneration

描述 TIMP3 在黄斑变性中的作用

基本信息

批准号：
10213739
负责人：
Ruchira Singh
金额：
$ 37.35万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-30 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10213739
关键词：
Affect Age related macular degeneration Atrophic Biogenesis Biological Models Biological Process Biology Cadaver Choroidal Neovascularization Complement Complement Activation Complement Membrane Attack Complex Complex Deposition Development Disease Drusen Electron Transport Complex III Enzymes Epithelial Physiology Equilibrium Event Extracellular Matrix Extracellular Matrix Proteins Eye Functional disorder Genes Genetic Transcription Histologic Histopathology Homeostasis Human Immune System Diseases Impairment In Vitro Inflammation Inflammatory Inherited Knock-in Knowledge Lead Link Lipids MMP14 gene MMP2 gene MMP9 gene Macular degeneration Mammalian Cell Matrix Metalloproteinases Mediating Modeling Molecular Mutation Outcome Pathogenesis Pathologic Pathology Pathway interactions Patients Pharmacotherapy Phenotype Physiology Play Production Proteins Regulation Retina Role Signal Transduction Site Sorsby&apos s fundus dystrophy Structure of retinal pigment epithelium TIMP3 gene TNF gene TNFRSF1A gene TNFRSF1B gene Testing Tight Junctions Variant Vascular Endothelial Growth Factors angiogenesis causal variant complement pathway cytokine disorder of macula of retina experimental study extracellular gene complementation induced pluripotent stem cell mouse model overexpression pigment epithelium-derived factor rare variant receptor retinal progenitor cell stem cell model targeted treatment

项目摘要

ABSTRACT Retinal pigment epithelium (RPE) cells in the retina are the major site of disease pathogenesis in numerous macular degenerative diseases. In a subset of these maculopathies, the primary pathological manifestations are localized to the RPE-extracellular matrix (RPE-ECM) complex, suggesting that ECM abnormalities contribute to the underlying disease mechanisms and RPE dysfunction in these maculopathies. In fact, mutations in TIMP3, a gene involved in ECM regulation, leads to Sorsby's fundus dystrophy (SFD), an inherited maculopathy with phenotypic alterations that mimic age-related macular degeneration (AMD). In addition, rare variants of TIMP3 are strongly linked to AMD development and TIMP3 accumulates underneath the RPE in both SFD and AMD. Together, these findings suggest that TIMP3 biology plays a central role in maintaining RPE-ECM homeostasis and that TIMP3 dysfunction contributes to pathophysiology of some maculopathies. However, the consequence of TIMP3 dysfunction on RPE physiology and its role in the development of macular degeneration has not been established. In this proposal, we will elucidate the molecular and pathological consequences of TIMP3 mutation in RPE cells using a patient-derived human- induced pluripotent stem cell (hiPSC)-RPE model of SFD. In preliminary studies, SFD hiPSC-RPE develops key pathological manifestation of the disease including, drusen formation and accumulation of TIMP3 in the extracellular matrix. Furthermore, consistent with TIMP3's biological function and pathophysiology of maculopathies affecting RPE-ECM integrity, SFD hiPSC-RPE displays altered expression of 1) pro-angiogenic TIMP3-target matrix metalloproteinases (MMPs; MMP2, MMP14), 2) TNF converting enzyme (TACE) and 3) complement pathway genes. These results support the hypothesis that TIMP3 dysfunction in SFD leads to dysregulation of ECM-turnover and activation of pro-angiogenic and pro-inflammatory signaling that consequently leads to drusen formation and RPE-mediated CNV. To test this hypothesis, we will assess the following predictions in our model system 1)) increased TIMP3 accumulation in ECM in SFD alters ECM composition/turnover and instigates complement activation in turn promoting drusen formation, 2) TIMP3 dysfunction leads to increased expression/activity of MMPs, MMP2 and MMP14, and thereby impaired barrier integrity and VEGF release by RPE cells in SFD and 3) TIMP3 dysfunction leads to TNF-induced pro- inflammatory signaling by RPE. These proposed experiments will provide clear answers the specific role of TIMP3 mutation/dysfunction in drusen biogenesis and RPE-mediated CNV. Ultimately, the knowledge gained in this study will help identify potential drug therapies for targeting TIMP3-induced RPE-ECM alterations in SFD and other maculopathies.!

抽象的视网膜中的视网膜色素上皮（RPE）细胞是许多疾病发病机制的主要部位。黄斑变性疾病。在这些黄斑病的一部分中，主要病理表现定位于 RPE-细胞外基质 (RPE-ECM) 复合体，表明 ECM 异常导致这些黄斑病的潜在疾病机制和 RPE 功能障碍。实际上， TIMP3（一种参与 ECM 调节的基因）的突变会导致索尔斯比眼底营养不良 (SFD)，这是一种遗传性黄斑病，其表型改变类似于年龄相关性黄斑变性（AMD）。在此外，TIMP3 的罕见变体与 AMD 的发展密切相关，并且 TIMP3 在下面积累 SFD 和 AMD 中的 RPE。总之，这些发现表明 TIMP3 生物学在维持 RPE-ECM 稳态，TIMP3 功能障碍导致某些疾病的病理生理学黄斑病。然而，TIMP3 功能障碍对 RPE 生理学的影响及其在黄斑变性的发展尚未确定。在本提案中，我们将阐明使用源自患者的人类 RPE 细胞中 TIMP3 突变的分子和病理后果 SFD的诱导多能干细胞（hiPSC）-RPE模型。在初步研究中，SFD hiPSC-RPE 开发该疾病的关键病理表现包括玻璃疣的形成和TIMP3在细胞外基质。此外，与 TIMP3 的生物学功能和病理生理学一致影响 RPE-ECM 完整性的黄斑病，SFD hiPSC-RPE 显示 1) 促血管生成的表达改变 TIMP3 靶基质金属蛋白酶（MMP；MMP2、MMP14）、2) TNF 转换酶 (TACE) 和 3) 补体途径基因。这些结果支持以下假设：SFD 中 TIMP3 功能障碍导致 ECM 周转失调以及促血管生成和促炎症信号传导的激活因此导致玻璃疣形成和 RPE 介导的 CNV。为了检验这个假设，我们将评估根据我们模型系统中的预测 1)) SFD 中 ECM 中 TIMP3 积累的增加会改变 ECM 组成/周转并刺激补体激活，进而促进玻璃疣形成，2) TIMP3 功能障碍导致 MMP、MMP2 和 MMP14 表达/活性增加，从而损害屏障 SFD 中 RPE 细胞的完整性和 VEGF 释放，以及 3) TIMP3 功能障碍导致 TNF 诱导的促- RPE 的炎症信号传导。这些提出的实验将提供明确的答案，具体作用玻璃疣生物发生和 RPE 介导的 CNV 中的 TIMP3 突变/功能障碍。最终获得的知识这项研究将有助于确定针对 SFD 中 TIMP3 诱导的 RPE-ECM 改变的潜在药物疗法和其他黄斑病。！