Delineating the role of TIMP3 in macular degeneration

描述 TIMP3 在黄斑变性中的作用

• 批准号: 10685477
• 负责人: Ruchira Singh
• 金额: $ 49.05万
• 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 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10685477
• 关键词: Age related macular degeneration; Area; Atrophic; Biological Models; Bruch' s basal membrane structure; CRISPR/Cas technology; Cell Line; Choroidal Neovascularization; Complex; Data; Deposition; Development; Disease; Disease model; Drusen; Event; Extracellular Matrix; Eye; Eye diseases; Functional disorder; Funding; Genes; Goals; HMGB1 gene; Homeostasis; Impairment; In Vitro; Inflammation; Intervention; LDL-Receptor Related Protein 1; Link; Lipids; Lipoproteins; Low Density Lipoprotein Receptor; Lysine; MMP2 gene; Macular degeneration; Matrix Metalloproteinases; Mediating; Modeling; Modification; Molecular; Mutation; Outcome Measure; Pathogenesis; Pathologic; Pathology; Patients; Pattern; Peptides; Pharmacotherapy; Phenotype; Phospholipase; Retinal Dystrophy; Role; Site; Sorsby' s fundus dystrophy; Sterility; Structure of retinal pigment epithelium; Study models; TIMP3 gene; Testing; Therapeutic; Vision; activation product; causal variant; disease mechanisms study; disorder of macula of retina; experimental study; extracellular; in vivo; induced pluripotent stem cell; inhibitor; knock-down; lipid metabolism; longitudinal analysis; macular dystrophy; monolayer; pharmacologic; rare variant; receptor for advanced glycation endproducts; small hairpin RNA; small molecule; stem cell model

ABSTRACT The retinal pigment epithelium-choriocapillaris (RPE-CC) complex is the primary site of disease pathogenesis in several eye diseases, including age-related macular degeneration (AMD) and related macular dystrophies, like Sorsby’s fundus dystrophy (SFD). Notably, sub-RPE accumulation of tissue inhibitor of metalloproteinase 3 (TIMP3) is a prominent feature of SFD/AMD. However, it is not known “how” sub-RPE TIMP3 accumulation promotes SFD/AMD pathology. This is partially because the RPE-CC is a functional composite making it difficult to study the contribution of spatial changes in RPE versus CC layer during disease development in vivo. Furthermore, the lack of a representative model of the RPE-CC in vitro has significantly impacted our ability to study RPE-CC interaction in vitro. Using induced pluripotent stem cells (iPSCs), we have developed an iPSC- RPE-CC model that recapitulate key features of both healthy and AMD/SFD eyes. Specifically, iPSC-RPE-CC shows evidence of fenestrated CC-like vasculature and Bruch’s membrane like ECM. Similarly, SFD iPSC-RPE- CC displays two central hallmarks of SFD/AMD, drusen and choroidal neovascularization (CNV)-like pathology. Notably, longitudinal analyses of control versus SFD iPSC-model showed that sub-RPE TIMP3 accumulation occurs relatively early and precedes maculopathy cellular events. Furthermore, our preliminary proof of concept studies shows that sub-RPE TIMP3 accumulation in the SFD iPSC model promotes pro-maculopathy cellular changes via dysregulated lipid metabolism and sterile inflammation. Based on these strong preliminary studies, the overall goal of this proposal is to establish the independent contribution of sub-RPE TIMP3 accumulation to AMD/SFD pathology development. We will perform spatial (RPE versus CC) and temporal (e.g., prior and after drusen formation) manipulation of TIMP3 expression/activity in i) SFD iPSC-RPE-CC and ii) control iPSC-RPE- CC cultures (iPSCs derived from healthy subjects with normal vision) to test the hypotheses that sub-RPE TIMP3 accumulation leads to dysregulated lipid metabolism and sterile inflammation and consequently i) drusen beneath the RPE monolayer and ii) CC atrophy and CNV. From a therapeutic standpoint, we will pharmacologically target dysregulated lipid metabolism and sterile inflammation in SFD/AMD iPSC models.

抽象的 视网膜色素上皮 - 胆管膜毛（RPE-CC）复合物是疾病发病机理的主要部位 几种眼科疾病，包括与年龄相关的黄斑变性（AMD）和相关的黄斑营养不良，例如 索斯比的眼底营养不良（SFD）。值得注意的是，金属蛋白酶3的组织抑制剂的亚RPE积累 （timp3）是SFD/AMD的突出特征。但是，尚不清楚“如何” sub-rpe timp3积累 促进SFD/AMD病理学。这部分是因为RPE-CC是一种功能性复合材料，使其变得困难 研究体内疾病发育过程中RPE与CC层的空间变化的贡献。 此外，缺乏体外RPE-CC的代表性模型显着影响了我们的能力 在体外研究RPE-CC相互作用。使用诱导的多能干细胞（IPSC），我们开发了一个IPSC- RPE-CC模型概括了健康和AMD/SFD眼睛的关键特征。具体来说，IPSC-RPE-CC 显示出染色的CC样脉管系统和Bruch膜（如ECM）的证据。同样，SFD IPSC-RPE- CC显示了SFD/AMD，DRUSEN和脉络膜新生血管形成（CNV）类似病理学的两个中心标志。 值得注意的是，对照与SFD IPSC模型的纵向分析表明，sub-rpe timp3积累 相对较早地发生，并先于刺激细胞事件。此外，我们的初步概念证明 研究表明，SFD IPSC模型中的子RPE Timp3积累促进了促肌病细胞 通过失调的脂质代谢和无菌注射的变化。基于这些强大的初步研究 该提案的总体目标是建立子RPE Timp3积累的独立贡献 AMD/SFD病理发展。我们将执行空间（RPE与CC）和临时性（例如，先前和之后 drusen的形成）操纵i）sfd ipsc-rpe-cc和ii）的timp3表达/活性 CC培养物（IPSC源自具有正常视力的健康受试者），以测试Sub-Rpe Timp3的假设 积累会导致脂质代谢和无菌感染失调，因此I） 在RPE单层和II）CC萎缩和CNV之下。从治疗的角度来看，我们将 在SFD/AMD IPSC模型中，药理学靶向失调的脂质代谢和无菌注射。

项目成果

Characterization of Human iPSC-RPE on a Prosthetic Bruch's Membrane Manufactured From Silk Fibroin.

• DOI: 10.1167/iovs.17-23157
• 发表时间: 2018-06-01
• 期刊: Investigative ophthalmology & visual science
• 影响因子: 4.4
• 作者: Galloway CA;Dalvi S;Shadforth AMA;Suzuki S;Wilson M;Kuai D;Hashim A;MacDonald LA;Gamm DM;Harkin DG;Singh R
• 通讯作者: Singh R

Extracellular vesicles: an emerging player in retinal homeostasis.

• DOI: 10.3389/fcell.2023.1059141
• 发表时间: 2023
• 期刊: FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY
• 影响因子: 5.5
• 作者: Chatterjee, Amit;Singh, Ruchira
• 通讯作者: Singh, Ruchira