miR-17-92 exosome treatment of stroke

miR-17-92 外泌体治疗中风

• 批准号: 8996733
• 负责人: MICHAEL CHOPP
• 金额: $ 32.48万
• 依托单位: HENRY FORD HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8996733
• 关键词: Axon; Biological; Biological Process; Brain; Cell Therapy; Cells; Characteristics; Communication; Data; Development; Electrophysiology (science); Essential Genes; Fiber; Gene Targeting; Genes; Health; Injury; Lasers; Lead; Lipids; Mediating; Mesenchymal; Methods; MicroRNAs; Microfluidics; Middle Cerebral Artery Occlusion; Molecular; Nervous System Physiology; Nervous System Trauma; Neuraxis; Neurodegenerative Disorders; Neurologic; PTEN gene; Pathway interactions; Play; Population; Rattus; Recovery of Function; Regulator Genes; Regulatory Element; Role; Slice; Stroke; Stromal Cells; Testing; Therapeutic Effect; Thrombospondin 1; Translations; Untranslated RNA; base; connective tissue growth factor; design; exosome; functional outcomes; genetic regulatory protein; improved; in vivo; innovation; intercellular communication; mTOR Signaling Pathway; microvesicles; nerve injury; nervous system disorder; neurological recovery; neurovascular; novel strategies; post stroke; prototype; research study; response; stroke therapy; stroke treatment; success

 DESCRIPTION (provided by applicant): Exosomes, small lipid microvesicles (30-150 nm), are active biological containers, which transport regulatory genes and proteins between cells and form a major biological communication conduit, facilitating a plethora of biological responses. The regulatory molecules contained in the exosomes include microRNAs (miRNAs), short (22-25 nt) non-coding RNAs which regulate gene translation and play primary roles in mediating a vast range of biological functions. In this proposal, based on strong preliminary data, we propose to manufacture a distinct exosome population which contains increased levels of the miR-17-92 cluster as a proof-of-principle and a mechanistic demonstration of a new method of treating stroke and possibly other neurological diseases and injury. We test the premise, that by modulating their miRNA content, exosomes can be designed to enhance plasticity of axons and thereby further promote neurological recovery post stroke. Success of this novel approach may lead to a new designer-based paradigm for the treatment of stroke and neurological disease. The following Specific Aims and associated Hypotheses are proposed: Specific Aim 1: To employ exosomes derived from multipotent mesenchymal stromal cells (MSCs) to treat stroke in order to enhance neurovascular remodeling and thereby, functional recovery post stroke. Hypothesis: Exosomes, derived from MSCs when administered to rats after stroke promote neurovascular remodeling which improves functional outcome. Specific Aim 2: To alter specific miRNAs contained within exosomes generated by MSCs as a means to enhance axonal plasticity and neurological recovery post stroke. Hypothesis: Administration of exosomes with increased miR-17-92 cluster to rats post stroke promotes axonal remodeling and enhances functional outcome. There are multiple layers of innovation in our application: we generate biological exosome carriers tailored for specific miRNAs; we use these exosomes to treat stroke, without the administration of exogenous cells; we employ electrophysiological methods, laser capture, fiber track tracing, a battery of neurological tests, and an array of novel approaches, e.g. microfluidic chambers, and ex vivo slice cultures, to mechanistically determine the molecular pathways of the target exosomes which mediate axonal outgrowth. Development of this designer exosome-based therapy, also serves as a prototype for capitalizing on the characteristics of exosomes to transport specific miRNAs and for the manufacture of designer exosomes. Developing a therapy for stroke that is exosome-based, opens up a wide variety of means to deliver targeted regulatory genes to enhance multifaceted aspects of central nervous system (CNS) plasticity and to amplify neurological recovery for neural injury and neurodegenerative diseases.

 描述（由申请人提供）：外泌体，小脂质微泡（30-150 nm），是活性生物容器，它在细胞之间运输调节基因和蛋白质，形成主要的生物通讯管道，促进大量的生物反应。外泌体中包含的调节分子包括微小RNA (miRNA)、短(22-25 nt)非编码RNA，它们调节基因翻译并在介导广泛的生物功能中发挥主要作用。在这项提案中，基于强有力的初步数据，我们建议制造一种独特的外泌体群体，其中含有增加的 miR-17-92 簇水平，作为治疗中风和可能的其他神经系统疾病和损伤的新方法的原理验证和机制论证。我们测试的前提是，通过调节 miRNA 含量，外泌体可以被设计来增强轴突的可塑性，从而进一步促进中风后的神经功能恢复。这种新方法的成功可能会带来一种新的基于设计者的治疗中风和神经系统疾病的范例。提出以下具体目标和相关假设： 具体目标 1：利用多能间充质基质细胞 (MSC) 衍生的外泌体治疗中风，以增强神经血管重塑，从而增强中风后的功能恢复。假设：源自 MSC 的外泌体给予中风后的大鼠，可促进神经血管重塑，从而改善功能结果。具体目标 2：改变 MSC 产生的外泌体中包含的特定 miRNA，作为增强轴突可塑性和中风后神经恢复的手段。假设：对中风后的大鼠施用含有增加的 miR-17-92 簇的外泌体可促进轴突重塑并增强功能结果。我们的应用有多个层面的创新：我们生成针对特定 miRNA 定制的生物外泌体载体；我们使用这些外泌体来治疗中风，无需施用外源细胞；我们采用电生理学方法、激光捕获、纤维轨迹追踪、一系列神经学测试以及一系列新颖的方法，例如微流体室和离体切片培养物，以机械地确定介导轴突生长的目标外泌体的分子途径。这种基于外泌体的设计疗法的开发，也可作为利用外泌体的特性来运输特定 miRNA 和制造设计外泌体的原型。开发一种基于外泌体的中风疗法，开辟了多种手段来传递靶向调节基因，以增强中枢神经系统（CNS）可塑性的多方面，并增强神经损伤和神经退行性疾病的神经恢复。