Defining oxidative stress induced changes in RPE that control RPE and photoreceptor degeneration

定义氧化应激诱导的 RPE 变化，控制 RPE 和光感受器变性

• 批准号: 10315588
• 负责人: Manas R Biswal
• 金额: $ 22.43万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10315588
• 关键词: Affect; Affinity Chromatography; Age; Age related macular degeneration; Atrophic; Binding; Biochemical; Biological Assay; Blindness; Bruch' s basal membrane structure; CASP1 gene; CCL2 gene; Cathepsins B; Cell Culture Techniques; Cell Surface Proteins; Cell surface; Cessation of life; Choroidal Neovascularization; Chronic; Confocal Microscopy; Data Set; Defect; Deposition; Development; Disease; Disease Progression; ERCC2 gene; Electron Microscopy; Enzyme-Linked Immunosorbent Assay; Event; Exudative age-related macular degeneration; Eye; Fluorescein-5-isothiocyanate; Foundations; Functional disorder; Gene Expression; Genes; Goals; Health; Immunofluorescence Immunologic; Immunohistochemistry; Impairment; In Vitro; Individual; Inflammasome; Inflammatory; Injury; Interleukin-1 alpha; Interleukin-18; Interleukin-6; Label; Lasers; LysoTracker; Lysosomes; Magic; Maintenance; Measures; Membrane Proteins; Mitochondria; Modeling; Molecular; Mus; Nonexudative age-related macular degeneration; Ovalbumin; Oxidative Stress; Pathogenesis; Pathway interactions; Pattern; Phagocytes; Phagocytosis; Phagolysosome; Phagosomes; Phenotype; Photoreceptors; Play; Process; Protein Isoforms; Proteins; Proteomics; Public Health; Reactive Oxygen Species; Reporting; Research; Retina; Retinal Degeneration; Retinal Photoreceptors; Retinoids; Reverse Transcriptase Polymerase Chain Reaction; Rhodopsin; Role; SOD2 gene; Scanning; Signal Pathway; Site; Stains; Streptavidin; Structure; Structure of retinal pigment epithelium; System; Systems Biology; Testing; Time; Training; Vacuole; Western Blotting; age related; base; career development; cytokine; differential expression; effective therapy; epithelial injury; geographic atrophy; improved; in vivo; loss of function; macula; mouse model; new therapeutic target; novel therapeutic intervention; novel therapeutics; photoreceptor cell outer segment; photoreceptor degeneration; prevent; recombinase-mediated cassette exchange; therapy development; tool; transcriptome sequencing

Summary Age-related macular degeneration (AMD) causes vision loss among many older individuals, and the retinal pigment epithelium (RPE) is thought to be a critical site of injury. Vision loss in AMD occurs due to photoreceptor degeneration and/or choroidal neovascularization. Geographic atrophy (GA), the advanced form of dry AMD, is characterized by the breakdown of RPE, choriocapillaris, and photoreceptors, especially in the macula. Lack of clear understanding of the molecular mechanisms of GA hinders the development of therapy. For lifelong maintenance of photoreceptors, RPE cells play an essential role in phagocytosis and degradation of tips shed from photoreceptor outer segments (POS). Photoreceptors and RPE cells are susceptible to injury from mitochondrial oxidative stress. The central goal of the project is to understand how photoreceptor degeneration occurs in GA. I hypothesize that oxidative stress impairs phagocytosis and lysosome function and ultimately activates inflammatory processes in RPE that stimulate geographic atrophy. I will test my hypothesis in RPE cell culture and in a new mouse model of age dependent RPE atrophy that was recently developed in our lab. In this model we used the cre/lox system to generate an RPE-specific deletion of Sod2, the mitochondrial gene for manganese superoxide dismutase (MnSOD). These mice develop a normal RPE but overtime the RPE has elevated oxidative stress resulting in phenotypic changes that are commonly observed in AMD, including RPE injury, loss of function and subsequent retinal degeneration. In the context of GA, I have following aims: (1) To characterize the impact of oxidative stress on phagocytosis, lysosomal function and inflammasome activation in RPE; (2) Identify molecular changes in RPE under oxidative stress. These studies will illuminate signaling pathways that drive photoreceptor and RPE loss and will provide a foundation to develop new therapeutic targets to prevent disease progression in AMD.

概括 与年龄相关的黄斑变性（AMD）导致许多老年人的视力丧失，并且 视网膜色素上皮（RPE）被认为是损伤的关键部位。 AMD的视力丧失 发生由于感光受体变性和/或脉络膜新生血管形成。地理 干燥AMD的高级形式萎缩（GA）的特征是RPE的崩溃， 绒毛膜毛细血管和感光体，尤其是在黄斑中。对 GA的分子机制阻碍了治疗的发展。用于终身维护 感光细胞，RPE细胞在吞噬和降解中起着至关重要的作用 来自光感受器外部段（POS）。感光细胞和RPE细胞易受损伤 来自线粒体氧化应激。该项目的核心目标是了解如何 光感受器变性发生在GA中。我假设氧化应激会损害 吞噬作用和溶酶体功能，并最终激活RPE的炎症过程 刺激地理萎缩。我将在RPE细胞培养和新鼠标中检验我的假设 我们实验室最近开发的年龄依赖性RPE萎缩模型。在这个模型中，我们 使用CRE/LOX系统生成SOD2的RPE特异性缺失，即线粒体基因 锰超氧化物歧化酶（MNSOD）。这些老鼠会形成正常的RPE，但加班 RPE具有升高的氧化应激，导致表型变化通常是 在AMD中观察到，包括RPE损伤，功能丧失和随后的视网膜变性。在 GA的背景，我有以下目的：（1）表征氧化应激对 RPE中的吞噬作用，溶酶体功能和炎性体激活； （2）识别分子 RPE在氧化应激下的变化。这些研究将照亮驱动的信号通路 感光器和RPE损失，并将为开发新的治疗靶标提供基础 预防AMD疾病进展。

项目成果

Carbon Dots Fabrication: Ocular Imaging and Therapeutic Potential.

• DOI: 10.3389/fbioe.2020.573407
• 发表时间: 2020
• 期刊: Frontiers in bioengineering and biotechnology
• 影响因子: 5.7
• 作者: Garner I;Vichare R;Paulson R;Appavu R;Panguluri SK;Tzekov R;Sahiner N;Ayyala R;Biswal MR
• 通讯作者: Biswal MR

Carbon Dot Nanoparticles: Exploring the Potential Use for Gene Delivery in Ophthalmic Diseases.

• DOI: 10.3390/nano11040935
• 发表时间: 2021-04-06
• 期刊: Nanomaterials (Basel, Switzerland)
• 作者: Biswal MR;Bhatia S
• 通讯作者: Bhatia S