Non-coding RNA regulation of sex differences in stroke

非编码RNA对中风性别差异的调节

• 批准号: 10605234
• 负责人: Creed Michael Stary
• 金额: $ 36.12万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10605234
• 关键词: 3' Untranslated Regions; Acute; Advanced Development; Affect; Animals; Apoptosis Inhibitor; Astrocytes; Biological; Biological Phenomena; Brain; Brain Injuries; Cause of Death; Cell Culture Techniques; Cell Death; Cell Survival; Cell physiology; Cells; Cessation of life; Clinical; Clinical Treatment; Clinical Trials; Data; Development; Disease; Elderly; Evolution; Experimental Models; Failure; Female; Fluorescent in Situ Hybridization; Functional disorder; Gene Expression Regulation; Gene Targeting; Generations; Genes; Glucose; Immunohistochemistry; In Vitro; Incidence; Injury; Intravenous; Investigation; Ischemia; Knowledge; Life; Link; Mediating; MicroRNAs; Middle Cerebral Artery Occlusion; Mitochondria; Molecular; Mus; Nerve Growth Factors; Neurodegenerative Disorders; Neurons; Outcome; Oxidative Phosphorylation; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Pharmacological Treatment; Play; Prevalence; Process; Proteins; RNA immunoprecipitation sequencing; RNA methylation; Reactive Oxygen Species; Regulation; Regulator Genes; Reperfusion Therapy; Resolution; Rodent; Role; SIRT1 gene; Sex Differences; Site; Small Interfering RNA; Source; Spirometry; Stroke; Testing; Therapeutic; Untranslated RNA; Woman; Work; age related; aged; brain cell; cell type; cohort; comparative; design; disability; fluorescence imaging; in vivo; in vivo evaluation; ischemic injury; long term recovery; male; men; mitochondrial dysfunction; novel; post stroke; pre-clinical; pre-clinical research; preclinical study; reelin protein; response; response to injury; sex; sexual dimorphism; stroke outcome; stroke therapy; translation factor; translational barrier; trend; young adult; 3' Untranslated Regions; Acute; Advanced Development; Affect; Animals; Apoptosis Inhibitor; Astrocytes; Biological; Biological Phenomena; Brain; Brain Injuries; Cause of Death; Cell Culture Techniques; Cell Death; Cell Survival; Cell physiology; Cells; Cessation of life; Clinical; Clinical Treatment; Clinical Trials; Data; Development; Disease; Elderly; Evolution; Experimental Models; Failure; Female; Fluorescent in Situ Hybridization; Functional disorder; Gene Expression Regulation; Gene Targeting; Generations; Genes; Glucose; Immunohistochemistry; In Vitro; Incidence; Injury; Intravenous; Investigation; Ischemia; Knowledge; Life; Link; Mediating; MicroRNAs; Middle Cerebral Artery Occlusion; Mitochondria; Molecular; Mus; Nerve Growth Factors; Neurodegenerative Disorders; Neurons; Outcome; Oxidative Phosphorylation; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Pharmacological Treatment; Play; Prevalence; Process; Proteins; RNA immunoprecipitation sequencing; RNA methylation; Reactive Oxygen Species; Regulation; Regulator Genes; Reperfusion Therapy; Resolution; Rodent; Role; SIRT1 gene; Sex Differences; Site; Small Interfering RNA; Source; Spirometry; Stroke; Testing; Therapeutic; Untranslated RNA; Woman; Work; age related; aged; brain cell; cell type; cohort; comparative; design; disability; fluorescence imaging; in vivo; in vivo evaluation; ischemic injury; long term recovery; male; men; mitochondrial dysfunction; novel; post stroke; pre-clinical; pre-clinical research; preclinical study; reelin protein; response; response to injury; sex; sexual dimorphism; stroke outcome; stroke therapy; translation factor; translational barrier; trend; young adult

Stroke remains the second leading cause of death worldwide and the primary cause of long-term disability in the US. Although pre-clinical studies have identified hundreds of potential drug agents, the only pharmacological treatment available remains early reperfusion with thrombolytics. A major factor for translational failure may be the overwhelming use of young male rodents in pre-clinical research, and lack of sex-specific design and analysis in clinical trials. Stroke is a sexually dimorphic disease with sex differences in incidence, prevalence, and outcome. Mitochondria mediate post-ischemia cell death in both males and females, but in different manners. Non-coding RNAs, including microRNAs (miRs), are upstream regulators of genes that regulate cell survival and mitochondrial function. In young adult male animals, miRs have been established as central regulators in the cellular response to stroke, however their role in females or aged animal cohorts represents a critical knowledge gap. Our prior studies (2, 4) and preliminary evidence suggest age-related differences between females and males in expression of select miRs and their targets following stroke. The overarching hypothesis of these Aims is that expression and function of miR-181a and miR-200c is a critical determinant of sexual dimorphism in stroke outcomes. To test this, we will first compare aged (20 month old) female and male mice using the middle cerebral artery occlusion (MCAO) model of experimental stroke, then compare primary female and male neuronal and astrocyte cultures utilizing in vitro ischemia. Preliminary evidence in aged animals reveals only a modest post- MCAO miR-181a response that is limited to females, while comparatively, a pronounced miR-200c response occurs in both sexes. Moreover, our preliminary evidence implies sex-differences in miR-181a and miR-200c gene targeting. In Aim 1, we will first compare protection with post-MCAO IV treatment of anti-miR-181a or anti- miR-200c in aged males and females. Then, we will assess the long-term cell-specific and sex-specific response in miR-181a and miR-200c to MCAO. Next, we will compare sustained post-MCAO miR-200c inhibition on long- term neuro-recovery between aged males and females, with the basis that preliminary evidence implies female- specific targeting of the neurotrophic protein reelin by miR-200c. In Aim 2, we will: 1) focus on the roles of miR- 181a and miR-200c in differences in regional (core versus penumbra) post-stroke disruption in mitochondrial function between aged males and females; and, 2) determine the roles of miR-181a and miR-200c in cell-type, and sex-specific alterations in oxidative phosphorylation and disruptions in cytosolic and mitochondrial Ca2+- handling following in vitro ischemia. In Aim 3 we will first define cell-type and sex-dependent differential gene targeting by miR-181a (Grp78 and XIAP) and miR-200c (XIAP, sirtuin-1, reelin). Finally we will determine whether observed differential gene targeting could be attributed to differences in RNA methylation, and whether this changes in response to injury. Delineating the molecular mechanisms that determine sexual dimorphism in stroke is necessary to overcome translational barriers and advance the development of novel stroke treatments.

中风仍然是全球死亡的第二大原因，也是长期残疾的主要原因 我们。尽管临床前研究已经确定了数百种潜在药物，但唯一的药理 可用的治疗仍然具有溶栓剂的早期再灌注。翻译失败的主要因素可能是 年轻的雄性啮齿动物在临床前研究中的压倒性用途以及缺乏性别的设计和分析 在临床试验中。中风是一种性二态性疾病，发病率，患病率和 结果。线粒体介导了男性和女性的缺血后细胞死亡，但以不同的方式进行。 包括microRNA（miR）在内的非编码RNA是调节细胞存活和的基因的上游调节剂 线粒体功能。在年轻的成年雄性动物中，MIR被确定为中央调节剂 细胞对中风的反应，但是它们在女性或老年动物队列中的作用代表了关键知识 差距。我们先前的研究（2，4）和初步证据表明，女性与年龄相关的差异 中风后选择的MIR及其靶标表达的雄性。这些目标的总体假设 miR-181a和miR-200c的表达和功能是中风中性二态性的关键决定因素 结果。为了测试这一点，我们将首先使用中大脑比较年迈的（20个月大）的雌鼠和雄性小鼠 实验中风的动脉闭塞（MCAO）模型，然后比较原发性女性和雌性神经元和 使用体外缺血的星形胶质细胞培养物。老年动物中的初步证据只揭示了一个适中的证据。 MCAO miR-181a响应仅限于女性，而相对的miR-200c反应 出现在男女中。此外，我们的初步证据意味着miR-181a和miR-200c中的性别差异 基因靶向。在AIM 1中，我们将首先将保护与MCAO后IV治疗抗MIR-181A或抗 - Mire-200c在老年男性和女性中。然后，我们将评估长期细胞特异性和性别的反应 在mir-181a和miR-200c中，到MCAO。接下来，我们将比较持续的mcao miR-200c抑制作用 老年男性和女性之间的术语神经恢复，初步证据暗示着女性 miR-200c对神经营养蛋白reelin的特异性靶向。在AIM 2中，我们将：1）关注mir的作用 181a和miR-200c在线粒体中触摸后区域（核心与半阴茎）的差异差异 老年男性和女性之间的功能； 2）确定miR-181a和miR-200c在细胞类型中的作用， 氧化磷酸化和细胞体和线粒体Ca2+ - 的氧化磷酸化以及性别特异性改变 - 处理以下体外缺血。在AIM 3中，我们将首先定义细胞类型和性别依赖性差异基因 由miR-181a（GRP78和XIAP）和miR-200c（XIAP，Sirtuin-1，reelin）靶向。最后，我们将确定是否 观察到的差异基因靶向可以归因于RNA甲基化的差异，以及这是否是 响应伤害的变化。描述决定性二态性的分子机制 中风对于克服翻译障碍并推动新型中风治疗的发展是必要的。