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: 10843511
• 负责人: STEVEN ROTH
• 金额: $ 5.16万
• 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/10843511
• 关键词: Acute; Address; Adjuvant; Affect; Alteplase; Animal Model; Anti-Inflammatory Agents; Antiinflammatory Effect; Apoptosis; Apoptotic; Attenuated; Basic Science; Biology; Biomimetics; Blindness; Blood-Retinal Barrier; Cardiac Surgery procedures; Case Series; Cell Death; Cell secretion; Cells; Central Retinal Artery Occlusion; Characteristics; Cytokine Gene; Cytoprotection; Dependence; Dermal; Diabetic Retinopathy; Disease; Economic Burden; Elderly; Emergency Situation; Engineering; Enrollment; Event; Eye; Face; Filler; Generations; Growth; Hyperbaric Oxygen; Hypoxia; Iatrogenesis; Immune response; In Vitro; Incidence; Inflammation; Inflammatory; Injections; Interruption; Intervention; Intravenous; Ischemia; Knowledge; Laboratories; Long-Term Care; Massage; Mediating; Mesenchymal Stem Cells; Methodology; MicroRNAs; Microglia; Modeling; Molecular; Muller' s cell; Natural regeneration; Neurons; Paracentesis; Pathway interactions; Patients; Permeability; Physiologic Intraocular Pressure; Play; Property; Public Health; Publishing; Reporting; Retina; Retinal Diseases; Retinal Ganglion Cells; Rodent Model; Role; Specificity; Stroke; System; Testing; Therapeutic; Translating; Vascular Diseases; Vascular Endothelial Cell; Vision; antagonist; anterior chamber; cell injury; cerebrovascular; clinical translation; combat; disability burden; effectiveness evaluation; experimental study; extracellular vesicles; falls; functional disability; functional mimics; gain of function; human disease; human model; immunoregulation; in vivo; innovation; ischemic injury; knock-down; loss of function; model development; nanovesicle; neuroinflammation; neuroprotection; novel; overexpression; precision medicine; preconditioning; prevent; protective effect; randomized, clinical trials; repaired; response; restoration; retinal damage; retinal ischemia; retinal neuron; stem cells; stroke model; success; thrombolysis; tool; transcriptome sequencing; translational medicine; vascular injury

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 re- sponses, 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 containing function-specific miRNA. Compared to MSCs, their EVs are non-immunogenic, non-tumorigenic, and modifiable for specific delivery modes. These characteristics render them ideal biomimetic agents fitting precision-based medicine. 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-inflam- mation. 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 miR-424/other key miRNAs in MSC-EVs will re-capitulate the anti-apoptosis and anti-inflammatory actions of H-EVs. We designate such EVs as Func- tionally Engineered EVs (FEEs). To facilitate clinical translation of MSC-EV therapy, we have identified key fun- damental 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? In Aim 1 we will produce FEEs overexpressing miR- 424 (FEE-424) and 146b (FEE-146b). We will evaluate the mechanisms of action of the FEEs, and their candi- dacy for generation of FEEs in retinal ganglion cells, microglia, Muller cells, and retinal vascular endothelial cells using loss and gain of function studies in models of simulated ischemia in vitro. These results will serve as a proof-of-principle model for development of FEEs for amelioration of cell damage in the retina. In Aim 2, FEEs containing miR-424 and -146b will be used to test specific targeting of anti-apoptotic and inflammatory mecha- nisms in a rodent model of CRAO. Overall, the 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. Innovations are cell-free therapy of retinal diseases, EV miR-mediated application-specificity, and direct determination of the impact of EVs on specific cells involved in retinal ischemic injury. Translational significance is the high likelihood of impacting novel molecular therapy. Underlying basic research significance is that the studies will enable vertical advancement of the field by determining mechanisms of actions of EVs in the retina.

中央视网膜动脉阻塞（CRAO）是一种眼科紧急情况，几乎没有经过验证的疗法。干 基于细胞的视网膜细胞置换是实现视网膜神经保护和到达的一种令人鼓舞的方法 保存视网膜疾病的视力。但是，由于局限 货物和异常生长需要一种替代的无细胞方法。电动汽车是纳米深头体， 当靶细胞内吞肠细胞时，触发特定的响应。在这里，电动汽车的microRNA（miRNA）货物 扮演关键角色。该建议针对使用含有工程的MSC-EV的视网膜功能恢复 功能特异性miRNA。与MSC相比，它们的电动汽车是非免疫原性的，非肿瘤的和可修改的 用于特定的交货模式。这些特征使它们理想地拟合基于精度的仿生剂 药品。我们的研究表明，电动汽车可以挽救遭受低氧或缺氧或 缺血，这是开始在CRAO中死亡的关键机制。我们还发现 MSC导致EVS（H-EV）具有增强的细胞保护特性，包括抗凋亡和抗炎症 - mation。 H-EV中过表达的许多miRNA在视网膜细胞中具有细胞保护特性。 我们的中心假设是msc-evs中miR-424/其他关键miRNA的靶向EV特异性表达 将重新列出H-EV的抗凋亡和抗炎作用。我们将EVS指定为func- 经过工程的电动汽车（费用）。为了促进MSC-EV治疗的临床翻译，我们确定了关键的有趣 - 大型知识差距：（1）EV miRNA及其抗凋亡特性之间的关系； （2）EV miRNA及其在视网膜中MSC-EV的抗炎作用中的作用； （3）可以增强MSC-EV 通过工程MiRNA货物进行有针对性的功能？在AIM 1中，我们将产生过表达mir-的费用 424（费用424）和146b（费用146b）。我们将评估费用的作用机制及其候选 DACY用于视网膜神经节细胞，小胶质细胞，毛毛细胞和视网膜血管内皮细胞的发电 在体外模拟缺血模型中使用功能研究的丧失和增益。这些结果将作为 原则上的原则模型，用于改善视网膜细胞损伤的费用。在AIM 2中，费用 包含miR-424和-146b将用于测试抗凋亡和炎症性机械的特定靶向 在Crao啮齿动物模型中的nism。总体而言，拟议的研究有望提供变革性的结果 因此，在缺血事件治疗CRAO之后，将MSC-EV进行了修改并进行视网膜保护作用。 创新是视网膜疾病的无细胞治疗，EV miR介导的应用特异性和直接的 确定电动汽车对视网膜缺血性损伤的特定细胞的影响。翻译意义 是影响新型分子疗法的很高可能性。基础研究意义是 研究将通过确定视网膜中电动汽车作用的机制来实现该领域的垂直进步。