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.

视网膜中央动脉阻塞（CRAO）是一种眼科急症， 治疗基于干细胞的视网膜细胞替代是一种非常令人鼓舞的方法， 视网膜神经保护和挽救视网膜疾病的视力。然而，有一些限制，包括 少数细胞整合，不良免疫反应和异常生长，替代无细胞 方法是必需的。EV是纳米囊泡体，当被靶细胞内吞时， 引发特定的反应。在这里，EV的microRNA（miRNA）货物起着关键作用。这 建议使用具有功能特异性的工程MSC-EV恢复视网膜功能 小RNA。我们的研究表明，电动汽车可以拯救视网膜细胞， 缺氧或缺血，这是CRAO中细胞死亡的关键机制。我们还发现 低氧预处理的MSCs产生了具有增强的细胞保护特性的EV（H-EV 包括抗凋亡和抗炎。在H-EV中过表达的一些miRNAs 对视网膜细胞有细胞保护作用。 我们的中心假设是，MSC-EV中关键miRNA的靶向EV特异性表达 将重新发挥H-EV的抗凋亡和抗炎作用。我们指定这样的 功能工程电动汽车（FEE）。为了促进MSC-EV治疗的临床转化， 我们发现了关键的知识空白：（1）EV miRNA与其抗凋亡的关系 （2）EV miRNA及其在视网膜中MSC-EV抗炎作用中的作用;（3） MSC-EV是否可以通过设计其miRNA货物来增强靶向功能？要求1 将产生过表达miR-424（FEE-424）和146 b（FEE-146 b）的FEE。我们将评估 它们的作用机制，以及它们在视网膜神经节细胞中产生FEE的候选资格， 小胶质细胞、Muller细胞和视网膜血管内皮细胞的功能丧失和获得研究 在体外模拟缺血。这些结果将作为开发的原理证明， FEE用于改善视网膜中的细胞损伤。在目标2中，含有miR-424和miR-146 b的FEE 将在啮齿动物模型中测试抗凋亡和炎症机制的特异性靶向， CRAO。预计拟议的研究将提供变革性的结果， 在缺血性事件后进行修饰并递送用于视网膜保护作用以治疗CRAO。 多样性补充方案将培训一名研究生进行数据收集和分析， 批判性地回顾文献，设计实验，并提交和撰写研究报告。 我们的目标是为这个经验，以提高学生的技能，在执行翻译的基本 科学研究，并确保有更多的机会使科学工作队伍多样化。