miR-17-92 exosome treatment of stroke

 描述(申请人提供)：Exosome，小的脂微囊(30-150 nm)，是活性的生物容器，在细胞之间运输调节基因和蛋白质，并形成一个主要的生物沟通渠道，促进大量的生物反应。外切体中包含的调控分子包括microRNAs(MiRNAs)和短的(22-25nt)非编码RNA，它们调节基因的翻译，并在广泛的生物学功能中发挥主要作用。在这项建议中，基于强大的初步数据，我们建议制造一个独特的外体群体，其中包含水平增加的miR-17-92簇，作为原则证明和治疗中风以及可能的其他神经疾病和损伤的新方法的机械演示。我们测试的前提是，通过调节其miRNA含量，外切体可以被设计成增强轴突的可塑性，从而进一步促进中风后的神经恢复。这一新方法的成功可能会带来一种基于设计者的治疗中风和神经疾病的新范式。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(MSCs)来源的外体治疗卒中，以促进神经血管重塑，从而促进卒中后功能恢复。假设：中风后给予大鼠骨髓间充质干细胞来源的外切体可促进神经血管重塑，从而改善功能结局。具体目标2：改变MSCs产生的外切体中包含的特定miRNAs，以此作为增强轴突可塑性和卒中后神经恢复的手段。假设：给卒中后的大鼠注射含有增加miR-17-92簇的外切体可促进轴突重塑并改善功能结果。我们的应用程序有多个层面的创新：我们产生为特定miRNAs量身定做的生物Exosome载体；我们使用这些Exosome治疗中风，而不使用外源细胞；我们使用电生理学方法、激光捕获、纤维跟踪跟踪、一系列神经学测试和一系列新方法，例如微流体室和体外切片培养，以机械方式确定介导轴突生长的目标Exosome的分子路径。这种设计的基于外显体的疗法的开发，也是利用外显体的特性来运输特定的miRNAs和制造设计外显体的原型。开发一种基于外体的中风治疗方法，开辟了多种手段来传递靶向调控基因，以增强中枢神经系统(CNS)多方面的可塑性，并扩大神经损伤和神经退行性疾病的神经康复。

 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.