Investigating the transfer of miRNAs by exosomes during inflammation

研究炎症过程中外泌体的 miRNA 转移

• 批准号: 9310060
• 负责人: Ryan M O'Connell
• 金额: $ 37.84万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9310060
• 关键词: Acute; Autoimmunity; Biochemical; Bone Marrow; CRISPR/Cas technology; Cardiovascular Diseases; Cells; Chimera organism; Chronic; Clinic; Clinical Research; Communication; Custom; Data; Dendritic Cells; Disease; Endotoxins; Engineering; Fractionation; Gene Targeting; Genes; Grant; Human; Immune; Immune response; Immune system; Impairment; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Injection of therapeutic agent; Light; Lipids; Malignant Neoplasms; Mediating; Methods; MicroRNAs; Molecular; Movement; Multivesicular Body; Mus; Nerve Degeneration; Obesity; Pathologic Processes; Phenotype; Physiological; Play; Process; Production; Radiation Chimera; Reporting; Role; Small RNA; Study models; Symptoms; System; Technology; Testing; Therapeutic; Therapeutic Uses; Tissues; Toxic effect; Translating; Vesicle; base; cell type; combat; exosome; experimental study; gene repression; human disease; immunoregulation; improved; in vivo; inhibitor/antagonist; insight; intercellular communication; knock-down; microbial; mouse model; novel; prevent; receptor mediated endocytosis; response; success; therapeutic miRNA

Abstract Mammalian microRNAs are critical regulators of several human diseases. As a result, therapeutic manipulation of miRNAs in diseased tissues has emerged as a promising approach to combating human disorders, and has had success in some settings. Yet, effective delivery of miRNAs or their inhibitors to many cell and tissue types continues to be a major challenge, and this underscores the need for improved methods of small RNA delivery to relevant cell types. Recent reports have demonstrated that miRNAs can be released from cells in small lipid vesicles called exosomes. In turn, the exosomes can deliver miRNAs to recipient cells, and this system is thought to constitute a novel form of intercellular communication. This process includes an endogenous miRNA delivery mechanism that is largely uncharacterized, yet could provide valuable insights into how therapeutic miRNA delivery can be improved and how inflammatory responses are regulated. Our preliminary data demonstrate that mouse bone marrow derived dendritic cells (BMDCs) produce exosomes that contain specific miRNAs, including the inflammatory regulators miR-155 and mIR-146a, that can be delivered to recipient immune cells and subsequently mediate target knockdown both in vitro and in vivo. This results in an altered response to endotoxin both in vitro and in vivo, which provides experimental evidence that exosome-transferred miRNAs provide a novel layer of regulating inflammation. Further, we have also been able to successfully load specific miRNA mimics into exosomes and demonstrate that they can be delivered to recipient cells in a functionally relevant manner. As a result of these findings, we hypothesize that exosomal miRNAs play novel regulatory roles during physiologically relevant inflammatory responses, and that through an improved understanding of this process, exosomes can ultimately be coopted to deliver specific miRNA cocktails in a therapeutically relevant manner. We will carry out the following specific aims to test these predictions. First, we will define the process of exosomal miRNA delivery to immune cells. Next, we will determine the functional relevance of exosomal miRNAs during inflammation using radiation chimeras and Rab27a/b DKO mice with impaired exosome production. Finally, we will engineer custom exosomes and test their impact on inflammatory disease. Together, this project will shed light on how exosome transfer of miRNAs is regulated, determine the role of this system during physiologically relevant inflammation, and begin to understand the translational potential of this novel process.

抽象的 哺乳动物microRNA是几种人类疾病的关键调节剂。结果，治疗 患病组织中miRNA的操纵已成为对抗人的有前途的方法 疾病，并在某些情况下取得了成功。然而，有效地将miRNA或其抑制剂传递给许多 细胞和组织类型仍然是一个重大挑战，这强调了改进方法的需求 小的RNA输送到相关的细胞类型。最近的报告表明，miRNA可以发布 来自小脂质囊泡中的细胞，称为外泌体。反过来，外泌体可以向受体细胞传递miRNA 并且该系统被认为构成了细胞间交流的一种新型形式。这个过程包括一个 内源miRNA递送机制在很大程度上没有特征，但可以提供有价值的见解 如何改善治疗性miRNA递送以及如何调节炎症反应。我们的 初步数据表明，小鼠骨髓衍生的树突状细胞（BMDC）产生外泌体 包含特定的miRNA，包括炎症调节剂miR-155和miR-146a，可以是 递送到受体免疫细胞中，随后介导体外和体内靶向敲低。这 导致对体外和体内内毒素的反应改变，这提供了实验证据，证明 外部转移的miRNA提供了调节炎症的新层。此外，我们也一直 能够成功地将特定的miRNA模拟加载到外泌体中，并证明它们可以传递到 受体细胞以功能相关的方式。由于这些发现，我们假设外泌体 miRNA在生理上相关的炎症反应中扮演新颖的调节作用，并通过 对这一过程的改进理解，外泌体最终可以使用特定的miRNA 鸡尾酒以治疗方式相关。我们将执行以下特定目标来测试这些 预测。首先，我们将定义外泌体miRNA递送到免疫细胞的过程。接下来，我们会的 使用辐射嵌合体和 RAB27A/B DKO小鼠外泌体产生受损。最后，我们将设计自定义外泌体和测试 它们对炎症性疾病的影响。该项目一起将阐明miRNA的外泌体转移 受调节，确定该系统在生理相关的炎症中的作用，然后开始 了解这一新过程的翻译潜力。