Delineating the role of TIMP3 in macular degeneration

描述 TIMP3 在黄斑变性中的作用

• 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
• 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' 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

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.!

摘要 视网膜中的视网膜色素上皮(RPE)细胞是许多疾病发病的主要部位 黄斑退行性疾病。在这些黄斑病变的一个子集中，主要的病理表现 定位于RPE-细胞外基质(RPE-ECM)复合体，提示ECM异常 有助于这些黄斑病变的潜在疾病机制和RPE功能障碍。事实上, 参与细胞外基质调节的基因TIMP3的突变导致索尔斯比眼底营养不良(SFD)，以及 遗传性黄斑病变，具有类似老年性黄斑变性(AMD)的表型改变。在……里面 此外，TIMP3的罕见变体与AMD的发生密切相关，TIMP3在下面积累 SFD和AMD中的RPE。综上所述，这些发现表明TIMP3生物学在 维持RPE-ECM动态平衡和TIMP3功能障碍在某些 黄斑病。然而，TIMP3功能障碍对RPE生理的影响及其在RPE中的作用 黄斑变性的发生尚未确定。在这项建议中，我们将澄清 使用患者来源的人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 本病的主要病理表现包括玻璃体形成和TIMP3在 细胞外基质。此外，与TIMP3的S的生物学功能和病理生理一致。 影响RPE-ECM完整性的黄斑病变，SFD HiPSC-RPE显示1)促血管生成的表达改变 TIMP3-靶向基质金属蛋白酶(MMPs；MMP2，MMP14)，2)肿瘤坏死因子转换酶(TACE)和3) 补体途径基因。这些结果支持了SFD中TIMP3功能障碍导致 ECM周转失调以及促血管生成和促炎信号的激活 从而导致玻璃体形成和RPE介导的CNV。为了检验这一假设，我们将评估 根据我们的模型系统中的预测，1)SFD患者ECM中TIMP3积累增加会改变ECM 合成/周转和激活补体进而促进玻璃疣的形成，2)TIMP3 功能障碍导致MMPs、MMP2和MMP14的表达/活性增加，从而损害屏障 SFD中RPE细胞的完整性和血管内皮生长因子的释放；3)TIMP3功能障碍导致肿瘤坏死因子诱导的促细胞因子释放。 RPE的炎症信号。这些拟议的实验将提供明确的答案，即 TIMP3突变/功能障碍与玻璃体生物发生和RPE介导的CNV归根结底，所获得的知识 这项研究将有助于确定针对TIMP3诱导的SFD RPE-ECM改变的潜在药物疗法 以及其他黄斑病变。