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

• 批准号: 10688145
• 负责人: STEVEN ROTH
• 金额: $ 31.76万
• 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: 2022-09-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10688145
• 关键词: Address; Adjuvant; Affect; Anti-Inflammatory Agents; Antiinflammatory Effect; Apoptosis; Athletic Injuries; Attenuated; Biology; Blindness; Cell Death; Cell Line; Cells; Cessation of life; 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' 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; clinically relevant; combat; cranium; cytokine; disability; exosome; experimental study; extracellular vesicles; functional mimics; glial activation; 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

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）是运动损伤视力丧失的主要原因，而汽车事故是 美国，目前没有治疗。我们的长期目标是发展安全，创新 打击TBI视力丧失的策略。轻度TBI（MTBI），即使没有直接创伤，也会导致残留 通过小胶质细胞和Müller细胞驱动的注射损害，以及残留神经节细胞丧失的功能障碍 （RGC）。间充质干细胞（MSC）细胞外蔬菜（EV）迅速作为干细胞出现 促进免疫调节，修复和再生的替代方案。将玻璃体注入的MSC EV 啮齿动物证明了RGC，Müller细胞和小胶质细胞的向流，并触发了特定的细胞反应， 电动汽车的微RNA（miRNA）发挥关键作用。 在这里，我们利用了我们已发布的初步数据，证明了MSC EV衰减小胶质 体外激活和RGC死亡。 MSC EV的抗凋亡和抗炎作用进一步增强 通过使用母体MSC的低氧预处理“增压” EV，产生“ H-EV”。 MSC细胞系 在遗传上改变为EV中的稳定过表达microRNA（即功能工程电动汽车或费用）， 费用模仿了H-EV的抗凋亡和抗炎作用。该建议针对中断 轻度TBI的视网膜损害，其中MSC-EV含有特定功能的miRNA，以减轻神经炎症 并执行神经元功能障碍和细胞死亡。我们的中心假设是针对性EV特异性的 关键miRNA的表达可以改善小胶质细胞和müller细胞驱动的注射，RGC和RGC的丧失 MTBI的随后视觉功能障碍。 AIM 1工程师EV模拟H-EV的功能，以减轻小胶质细胞和Müller细胞激活和 体外视网膜神经元死亡。我们假设费用增强了抗炎和抗凋亡的影响 MSC-EV和模拟H-EV。 H-EV中最高表达的miR中的两个将被研究为相比 在活化的视网膜小胶质细胞，RGC和Müller细胞中暴露于谷氨酸的MSC EV，H-EV和对照。 识别费用和H-EV的作用机制将为治疗精度提供新的途径 MTBI视觉功能障碍的医学。 AIM 2决定了与临床相关的小鼠中费用的体内抗炎和抗凋亡作用 MTBI模型。我们将测试以下假设，即给出的视网膜时给予玻璃体的费用 mtbi之后。 MTBI使用50 psi的视网膜功能，凋亡，谷氨酸水平和炎症介质 对颅骨的爆炸将进行定量比较，并在用H-处理的组中鉴定出细胞特异性作用 电动汽车，费用，MSC EV和控制。 预计拟议的研究将通过修改MSC-EV为保护行动提供新的结果 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。