VRC: Engineered extracellular vesicles for mild TBI-induced retinal injury

VRC：工程细胞外囊泡治疗轻度 TBI 引起的视网膜损伤

基本信息

批准号：
10598277
负责人：
STEVEN ROTH
金额：
$ 33.76万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10598277
关键词：
Address Adjuvant Affect Anti-Inflammatory Agents Antiinflammatory Effect Apoptosis Athletic Injuries Attenuated Biology Blindness Cell Death Cell Line Cells Cytoprotection Data Diabetic Retinopathy Engineering Exposure to Eye Functional disorder Genes Glutamates Goals Health Hypoxia In Vitro Inflammation Inflammation Mediators Inflammatory Injury Interruption Knowledge Laboratories Left Lentivirus Vector Mesenchymal Stem Cells Methodology MicroRNAs Microglia Muller&apos s cell Natural regeneration Neurodegenerative Disorders Neuronal Dysfunction Parents Pathway interactions Play Precision therapeutics Public Health Publishing Retina Retinal Diseases Retinal Ganglion Cells Rodent Rodent Model Role Specificity Sports Strategic Planning Stress Testing Therapeutic Translating Trauma Traumatic Brain Injury Tropism United States Visual antagonist automobile accident base clinically relevant combat cranium cytokine disability exosome experimental study extracellular vesicles human disease human model immunoregulation improved in vivo innovation knock-down loss of function mild traumatic brain injury mouse model neuroinflammation neuron loss novel overexpression precision medicine preconditioning prevent repaired response retinal damage retinal neuron stem cells stroke model transcriptome sequencing translational medicine visual dysfunction

项目摘要

Traumatic brain injury (TBI) is the leading cause of vision loss in sports injury, and automobile accidents in the United States, for which there currently is no treatment. Our long-term objective is to develop safe, innovative strategies to combat vision loss from TBI. Mild TBI (mTBI), even without direct trauma to the eye, results in retinal damage via microglia and Müller cell-driven inflammation, and dysfunction from loss of retinal ganglion cells (RGCs). Mesenchymal stem cell (MSC) extracellular vesicles (EVs) are rapidly emerging as a stem cell alternative that promote immunomodulation, repair, and regeneration. MSC EVs injected into the vitreous in rodents demonstrated tropism for RGCs, Müller cells, and microglia, and triggered specific cellular responses, with micro RNA (miRNA) from the EVs playing a key role. Here we capitalize upon our published and preliminary data demonstrating that MSC EVs attenuate microglial activation and RGC death in vitro. Anti-apoptosis and anti-inflammatory effects of MSC EVs were further boosted by “supercharging” the EVs using hypoxic preconditioning of the parent MSCs, yielding “H-EVs.” MSC cell lines were genetically altered to stably overexpress microRNAs in EVs (i.e., Functionally-Engineered EVs, or FEEs), and FEEs mimicked the anti-apoptosis and anti-inflammatory action of H-EVs. This proposal targets interrupting retinal damage from mild TBI with MSC-EVs containing function-specific miRNA to attenuate neuro-inflammation and accompanying neuronal dysfunction and cell death. Our central hypothesis is that targeted EV-specific expression of key miRNAs ameliorates microglia and Müller cell-driven inflammation, and loss of RGCs and subsequent visual dysfunction from mTBI. Aim 1 engineers EVs mimicking functionality of H-EVs to attenuate microglia and Müller cell activation and retinal neuron death in vitro. We hypothesize that FEEs enhance anti-inflammatory and anti-apoptosis effects of MSC-EVs and mimic H-EVs. Two of the highest expressed miRs in H-EVs will be studied as FEEs in comparison to MSC EVs, H-EVs, and controls in activated retinal microglia, and RGCs and Müller cells exposed to glutamate. Identifying mechanisms of action of FEEs and H-EVs will provide a novel pathway to therapeutic precision medicine for visual dysfunction in mTBI. Aim 2 determines in vivo anti-inflammatory and anti-apoptosis actions of FEEs in a clinically-relevant mouse model of mTBI. We will test the hypothesis that FEEs administered into the vitreous rescue the retina when given after mTBI. Retinal function, apoptosis, glutamate levels, and inflammatory mediators post mTBI using 50 psi blast to the cranium will be quantitatively compared, and cell-specific effects identified in groups treated with H- EVs, FEEs, MSC EVs, and controls. The proposed studies are expected to provide new results with MSC-EVs modified for protective action to treat the retinal cells affected by TBI, and provide novel potential therapy for other neurodegenerative disorders.

创伤性脑损伤 (TBI) 是导致运动损伤和汽车事故中视力丧失的主要原因美国目前尚无治疗方法，我们的长期目标是开发安全、创新的药物。对抗 TBI（mTBI）导致的视力丧失的策略，即使没有对眼睛造成直接创伤，也会导致视网膜损伤。小胶质细胞和穆勒细胞驱动的炎症造成的损伤，以及视网膜神经节细胞损失造成的功能障碍 (RGC) 间充质干细胞 (MSC) 细胞外囊泡 (EV) 正在迅速成为干细胞。促进注射到玻璃体中的 MSC EV 的免疫调节、修复和再生的替代方案。啮齿动物表现出对 RGC、Müller 细胞和小胶质细胞的趋向性，并引发特定的细胞反应， EV 中的微小 RNA (miRNA) 发挥着关键作用。在这里，我们利用已发表的初步数据证明 MSC EV 可以减弱小胶质细胞 MSC EVs 的体外激活和 RGC 死亡进一步增强。通过对亲本 MSC 进行缺氧预处理，对 EV 进行“增压”，从而产生“H-EV 细胞系”。经过基因改造，可在 EV 中稳定过度表达 microRNA（即功能工程 EV 或 FEE）， FEE 模仿了 H-EV 的抗细胞凋亡和抗炎作用。该提案的目标是中断。使用含有功能特异性 miRNA 的 MSC-EV 减轻轻度 TBI 引起的视网膜损伤，以减轻神经炎症以及伴随的神经元功能障碍和细胞死亡，我们的中心假设是针对 EV 特异性。关键 miRNA 的表达可改善小胶质细胞和 Müller 细胞驱动的炎症以及 RGC 和 RGC 的损失 mTBI 导致的后续视觉功能障碍。 Aim 1 工程师设计了模仿 H-EV 功能的 EV，以减弱小胶质细胞和 Müller 细胞的激活，并我们发现 FEE 增强了视网膜神经元的抗炎和抗凋亡作用。 MSC-EV 和模拟 H-EV 中 H-EV 中表达最高的两种 miR 将作为 FEE 进行研究进行比较。 MSC EV、H-EV 以及暴露于谷氨酸的激活的视网膜小胶质细胞、RGC 和 Müller 细胞中的对照。确定 FEE 和 H-EV 的作用机制将为实现治疗精准度提供新途径治疗 mTBI 视觉功能障碍的药物。目标 2 确定 FEE 在临床相关小鼠体内的抗炎和抗凋亡作用我们将检验以下假设：将 FEE 注射到玻璃体中可挽救视网膜。 mTBI 后使用 50 psi 测量视网膜功能、细胞凋亡、谷氨酸水平和炎症介质。将定量比较颅骨爆炸，并确定 H-治疗组中的细胞特异性效应 EV、FEE、MSC EV 和控件。拟议的研究预计将提供修改后的 MSC-EV 的新结果，以起到保护作用治疗受 TBI 影响的视网膜细胞，并为其他神经退行性疾病提供新的潜在疗法。