Role of Astrocytic miR-20a-3p as a Potential Therapeutic for Ischemic Stroke

星形细胞 miR-20a-3p 作为缺血性中风的潜在治疗药物的作用

• 批准号: 10675024
• 负责人: Taylor Branyan
• 金额: $ 3.65万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10675024
• 关键词: Adenoviruses; Affect; Age; Aging; Animal Model; Apoptotic; Astrocytes; Attenuated; Behavioral Assay; Bioinformatics; Biological Assay; Cell Hypoxia; Cell Survival; Cells; Chromatin; Corpus striatum structure; Culture Media; Data; Development; Disease; Down-Regulation; Drug Delivery Systems; Enzyme-Linked Immunosorbent Assay; Epigenetic Process; Estrogen Replacements; Estrogens; Exhibits; Female; Gene Transfer; Gene Transfer Techniques; Genes; Genetic Transcription; Genome; Glial Fibrillary Acidic Protein; Glutamates; Goals; Hour; Human; Hypoxia; Impairment; In Vitro; Infarction; Inflammation Mediators; Inflammatory; Injections; Ischemia; Ischemic Stroke; Link; Mediating; Menopause; MicroRNAs; Middle Cerebral Artery Occlusion; Mitochondria; Modeling; Modernization; Modification; Neuroprotective Agents; Oligonucleotides; Organelles; Outcome; Ovarian hormone; Perimenopause; Peripheral; Population; Postmenopause; Premenopause; Production; Proteins; Quantitative Reverse Transcriptase PCR; RNA; Rattus; Recombinant adeno-associated virus (rAAV); Recovery; Research; Risk; Role; Scientist; Sensorimotor functions; Severities; Stroke; Supplementation; System; Testing; Therapeutic; Time; Woman; Women' s Group; age related; brain cell; career; clinically relevant; cytokine; experience; extracellular vesicles; histone methylation; improved; in silico; in vivo; insight; member; men; middle age; neuroprotection; neurotoxic; novel therapeutics; post stroke; pre-clinical research; promoter; reproductive; senescence; sex; stroke incidence; stroke model; stroke outcome; stroke risk; therapeutic development; therapeutic target; translational potential; Adenoviruses; Affect; Age; Aging; Animal Model; Apoptotic; Astrocytes; Attenuated; Behavioral Assay; Bioinformatics; Biological Assay; Cell Hypoxia; Cell Survival; Cells; Chromatin; Corpus striatum structure; Culture Media; Data; Development; Disease; Down-Regulation; Drug Delivery Systems; Enzyme-Linked Immunosorbent Assay; Epigenetic Process; Estrogen Replacements; Estrogens; Exhibits; Female; Gene Transfer; Gene Transfer Techniques; Genes; Genetic Transcription; Genome; Glial Fibrillary Acidic Protein; Glutamates; Goals; Hour; Human; Hypoxia; Impairment; In Vitro; Infarction; Inflammation Mediators; Inflammatory; Injections; Ischemia; Ischemic Stroke; Link; Mediating; Menopause; MicroRNAs; Middle Cerebral Artery Occlusion; Mitochondria; Modeling; Modernization; Modification; Neuroprotective Agents; Oligonucleotides; Organelles; Outcome; Ovarian hormone; Perimenopause; Peripheral; Population; Postmenopause; Premenopause; Production; Proteins; Quantitative Reverse Transcriptase PCR; RNA; Rattus; Recombinant adeno-associated virus (rAAV); Recovery; Research; Risk; Role; Scientist; Sensorimotor functions; Severities; Stroke; Supplementation; System; Testing; Therapeutic; Time; Woman; Women' s Group; age related; brain cell; career; clinically relevant; cytokine; experience; extracellular vesicles; histone methylation; improved; in silico; in vivo; insight; member; men; middle age; neuroprotection; neurotoxic; novel therapeutics; post stroke; pre-clinical research; promoter; reproductive; senescence; sex; stroke incidence; stroke model; stroke outcome; stroke risk; therapeutic development; therapeutic target; translational potential

Project Summary The goal of this project is to test a therapeutic compound that has the potential to improve stroke outcomes in older females. It is critical to focus on this demographic because studies show that after menopause, women have greater risk and severity of ischemic stroke. Previous studies from the sponsor's lab have shown that reproductively senescent (acyclic) (RS) female rats have larger infarct and worse sensorimotor impairments than younger females. Moreover, estrogen replacement to RS females exacerbates stroke impairment. Thus this animal model replicates key features of stroke-related consequences seen in the human population, resulting in a model with translational potential, in which neurotherapeutic compounds can be tested. Prior research from the sponsor's lab has shown that astrocytes from reproductively senescent (RS) female rats exhibit age-related changes that contribute to poorer recovery after stroke. This includes a reduced capacity for glutamate clearance, decreased secretion of trophic factors, and increased release of inflammatory cytokines, compared to younger females. Furthermore, these aging astrocytes also display reduced histone methylation of the H3K4 chromatin region, resulting in less transcription of several genes including the microRNA mir-17-92 cluster. MicroRNAs have recently gained popularity as potential therapeutics for a variety of conditions, including ischemic stroke. Preliminary data for this proposal shows that miR-20a-3p, a member of the mir-17-92 cluster is dramatically down regulated in astrocytes of middle-aged females. Furthermore, iv injection of miR-20a-3p mimics four hours after middle cerebral artery occlusion (MCAo) decreased infarct volume and attenuated sensorimotor impairment in middle-aged females, suggesting that the miR-20a-3p could be mechanistically linked to stroke outcomes. This raises the intriguing possibility that in younger females, endogenous astrocyte-derived miR- 20a-3p may be `distributed' to other cells to improve stroke outcomes. This hypothesis will be tested in the following aims: Aim 1 will determine whether miR-20a-3p alters the composition of astrocytice extracellular vesicles (EVs) and that miR-20a-3p modified EVs will confer neuroprotection. Astrocytes have been shown to release neuroprotective factors, organelles and RNA translational machinery through EVs after stroke. In silico analysis predicts that miR-20a-3p may regulate inflammatory mediators and mitochondrial genes. Thus, miR- 20a-3p could drastically change the composition of the EVs. In aim 2, replication-deficient adenovirus constructs will be used to increase miR-20a-3p levels only in astrocytes and determine whether this recapitulates the neuroprotective effect of iv miR-20a-3p. Using clinically relevant stroke models, a battery of behavioral assays, and modern gene transfer techniques, this proposal will critically examine whether modification of the aging astrocyte will promote neuroprotection.

项目摘要 该项目的目的是测试具有改善中风结果的治疗化合物 年长的女性。关注这一人群至关重要，因为研究表明，更年期后，女性 缺血性中风的风险和严重程度更大。赞助商实验室的先前研究表明 生殖性衰老（无环）（RS）雌性大鼠的梗塞较大，感觉运动障碍较差 比年轻的女性。此外，雌激素替代RS女性加剧了中风障碍。因此 该动物模型复制了人口中与中风相关后果的关键特征， 导致具有转化电位的模型，可以测试神经治疗化合物。 赞助商实验室的先前研究表明，生殖性衰老（RS）雌性大鼠的星形胶质细胞 表现出与年龄相关的变化，导致中风后恢复较差。这包括降低的能力 谷氨酸清除，营养因子的分泌减少以及炎症细胞因子的释放增加， 与年轻的女性相比。此外，这些老化的星形胶质细胞还显示出降低的组蛋白甲基化 H3K4染色质区域，导致几种基因的转录较少，包括microRNA miR-17-92 簇。 MicroRNA最近作为各种疾病的潜在疗法广受欢迎， 包括缺血性中风。 该提案的初步数据表明，miR-17-92群集的miR-20a-3p是急剧 在中年女性的星形胶质细胞中调节。此外，静脉注射miR-20a-3p模拟四个 中大脑中动脉阻塞（MCAO）后数小时减少梗塞体积并减弱感觉运动 中年女性的损害，表明miR-20a-3p可能与中风有关 结果。这增加了一个有趣的可能性，即在年轻的女性中，内源性星形胶质细胞衍生的mir- 20A-3P可以“分布”到其他细胞中，以改善中风结果。该假设将在 以下目的：AIM 1将确定miR-20a-3p是否改变了星形胶质细胞的组成 囊泡（EV）和miR-20a-3p修饰的电动汽车将赋予神经保护作用。星形胶质细胞已显示为 释放神经保护因子，细胞器和RNA翻译机械通过中风后的电动汽车。在计算机中 分析预测，miR-20a-3p可能调节炎症介质和线粒体基因。因此，mir- 20A-3P可以大大改变电动汽车的组成。在AIM 2中，复制缺陷腺病毒 构造仅在星形胶质细胞中仅用于增加miR-20a-3p水平，并确定是否 概括IV miR-20a-3p的神经保护作用。使用临床相关的中风模型，一电池 行为测定和现代基因转移技术，该提案将批判性地检查是否是否 老化星形胶质细胞的修饰将促进神经保护作用。