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：
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
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/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&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 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

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&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 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

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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模仿H-EV的功能，以减轻小胶质细胞和Müller细胞的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。