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.

总结 视网膜相关性黄斑变性（AMD）在许多老年人中引起视力丧失， 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 由于光感受器变性和/或脉络膜新生血管形成而发生。地理 萎缩（GA），干性AMD的晚期形式，特征在于RPE的破坏， 脉络膜毛细血管和光感受器，尤其是黄斑。缺乏对 GA的分子机制阻碍了治疗的发展。终身维护 作为光感受器，RPE细胞在吞噬和降解脱落的尖端中起重要作用。 光感受器外节（POS）。光感受器和RPE细胞易受损伤 来自线粒体氧化应激。该项目的中心目标是了解如何 在GA中发生光感受器退化。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中的炎症过程， 刺激地图状萎缩。我将在视网膜色素上皮细胞培养和一只新的小鼠中检验我的假设 年龄依赖性RPE萎缩的模型，这是我们实验室最近开发的。在这个模型中，我们 使用cre/lox系统来产生RPE特异性Sod 2缺失，Sod 2是用于 锰超氧化物歧化酶（MnSOD）。这些老鼠的视网膜色素上皮细胞正常， RPE具有升高的氧化应激，导致表型变化， 在AMD中观察到的视网膜变性包括RPE损伤、功能丧失和随后的视网膜变性。在 本论文的主要目的是：（1）研究氧化应激对细胞凋亡的影响， RPE细胞吞噬功能、溶酶体功能和炎性小体激活;（2）分子鉴定 氧化应激下RPE的变化。这些研究将阐明信号通路， 光感受器和RPE的损失，并将提供基础，开发新的治疗目标， 预防AMD的疾病进展。

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