Mesenchymal stem cell extracellular vesicles for ischemic retinal damage

间充质干细胞胞外囊泡治疗缺血性视网膜损伤

• 批准号: 10766045
• 负责人: STEVEN ROTH
• 金额: $ 1.72万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10766045
• 关键词: Acute; Anti-Inflammatory Agents; Apoptosis; Apoptotic; Basic Science; Blindness; Cell Death; Cell secretion; Cells; Central Retinal Artery Occlusion; Cytoprotection; Data Analyses; Data Collection; Development; Emergency Situation; Engineering; Ensure; Event; Experimental Designs; Funding; Generations; Goals; Growth; Hypoxia; Immune response; In Vitro; Inflammation; Inflammatory; Ischemia; Knowledge; Mesenchymal Stem Cells; MicroRNAs; Microglia; Muller' s cell; Play; Property; Public Health; Reporting; Research; Research Design; Retina; Retinal Diseases; Retinal Ganglion Cells; Review Literature; Rodent Model; Role; Students; Testing; Therapeutic; Training; Vascular Endothelial Cell; Vision; Writing; biological research; cell injury; clinical translation; experience; extracellular vesicles; gain of function; graduate student; loss of function; nanovesicle; neuroprotection; overexpression; preconditioning; prevent; response; restoration; retinal damage; retinal ischemia; skills; stem cells; success; tool

Central retinal artery occlusion (CRAO) is an ophthalmological emergency with few proven therapies. Stem cell-based retinal cell replacement is a highly encouraging approach to achieve retinal neuroprotection and to save vision in retinal diseases. However, with limitations including few cells integrated, adverse immune responses, and aberrant growth, an alternative cell-free approach is required. EVs are nano-vesicular bodies that, when endocytosed by target cells, trigger specific responses. Here, the microRNA (miRNA) cargo of the EVs plays a key role. This proposal targets restoration of retinal function using engineered MSC-EVs with function-specific miRNA. Our studies indicate that EVs can rescue retinal cells that have been acutely subjected to hypoxia or ischemia, the key mechanism that starts cells dying in CRAO. We also found that hypoxic preconditioning of MSCs resulted in EVs (H-EVs) with enhanced cytoprotective properties including anti-apoptosis and anti-inflammation. A number of miRNAs overexpressed in the H-EVs have cytoprotective properties in retinal cells. Our central hypothesis is that targeted EV-specific expression of key miRNAs in MSC-EVs will re-capitulate the anti-apoptosis and anti-inflammatory actions of H-EVs. We designate such EVs as Functionally Engineered EVs (FEEs). To facilitate clinical translation of MSC-EV therapy, we identified key knowledge gaps: (1) The relationship between EV miRNA and its anti-apoptotic properties; (2) EV miRNA and its role in anti-inflammatory actions of MSC-EVs in retina; and (3) Can MSC-EVs be enhanced for targeted functionality by engineering their miRNA cargo? Aim 1 will produce FEEs overexpressing miR-424 (FEE-424) and 146b (FEE-146b). We will evaluate their mechanisms of action, and their candidacy for generation of FEEs in retinal ganglion cells, microglia, Muller cells, and retinal vascular endothelial cells using loss and gain of function studies in simulated ischemia in vitro. These results will serve as proof-of-principle for development of FEEs for amelioration of cell damage in the retina. In Aim 2, FEEs containing miR-424 and -146b will test specific targeting of anti-apoptotic and inflammatory mechanisms in a rodent model of CRAO. Proposed studies are expected to provide transformative results whereby MSC-EVs are modified and delivered for retinal protective action after the ischemic event to treat CRAO. The Supplement for Diversity will train a graduate student in data collection and analysis, reviewing literature critically, designing experiments, and presenting and writing research reports. The goal is for this experience to increase the student’s skills in performing translational basic science research and to ensure more opportunities for diversification of the scientific workforce.

中央视网膜动脉cocclusion（CRAO）是眼科紧急事件，可证明 疗法。 视网膜神经保护并保存视网膜疾病的视力。 很少有细胞整合，不良免疫反应和异常生长，一种无细胞 需要方法。 触发特定的响应。 提案目标使用具有特定功能的发动机MSC-EV进行视网膜功能的重新安排 miRNA。 对于低氧或缺血，恒星在CRAO中的关键机制也发现。 MSC的低氧预处理导致EVS（H-EV）具有增强的细胞保护特性 包含抗凋亡抗炎。 在视网膜细胞中具有细胞保护特性。 我们的中心假设是MSC-EV中关键miRNA的靶向EV特异性表达 我们将重新分配H-EV的抗凋亡抗炎作用 电动汽车作为功能设计的电动汽车（费用）。 我们确定了关键知识差距：（1）EV miRNA与抗凋亡之间的关系 （2）EV miRNA及其在视网膜中MSC-EV的抗炎作用中的作用； MSC-EV可以通过工程货物1来增强忘记的功能吗？ 将产生过表达mir-424（费用-424）和146b（费用146b）的费用 它们的作用机理以及在视网膜神经节细胞中产生的候选者， 使用功能研究的损失和增益 在体外模拟缺血中。 在AIM 2中改善细胞损伤的费用 将测试啮齿动物模型中抗凋亡和炎症机制的特定靶向 CRAO。 缺血性事件后，对视网膜保护器保护行动进行了修改和交付，以信任CRAO。 多样性供应量将培训研究和分析的研究生， 批判性地审查文献，设计实验以及介绍和写作研究报告。 目的是使执行力提高学生在执行翻译基本方面的技能 科学研究并确保更多的科学劳动力多样化机会。