Mechanisms for generating cargo specificity in inflammasome-mediated exosome secretion

炎症小体介导的外泌体分泌中产生货物特异性的机制

• 批准号: 10818688
• 负责人: Ann Lynn Wozniak
• 金额: $ 38.75万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-21 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10818688
• 关键词: AGFG1 gene; Adaptor Signaling Protein; Affect; Anti-Inflammatory Agents; Antigen Presentation; Binding; Biogenesis; Biological; Biological Process; CASP1 gene; Cell Communication; Cell surface; Cells; Complex; Data; Disease; Dynein ATPase; Endosomes; Event; Goals; Guanosine Triphosphate Phosphohydrolases; Human; Immune response; In Vitro; Inflammasome; Inflammatory; Inflammatory Response; Kinesin; Knowledge; Link; Liver; Liver diseases; Macrophage; Measures; Mediating; Membrane; MicroRNAs; Microtubules; Modification; Molecular Motors; Movement; Multivesicular Body; Outcome; Parents; Pathogenicity; Pathologic; Pathway interactions; Patients; Physiological; Physiological Processes; Play; Positioning Attribute; Process; Production; Proteins; Proteolysis; RNA; RNA-Binding Proteins; Recruitment Activity; Regulation; Research; Role; Route; Site; Sorting; Specificity; Stimulus; Testing; Therapeutic; Tissue Sample; Ubiquitination; Vesicle; Virus; Work; cell type; exosome; human disease; in vivo; intercellular communication; lysosomal proteins; monocyte; mouse model; nanosized; novel; patient population; programs; protein protein interaction; rab7 protein; response; tool; trafficking; virtual

PROJECT SUMMARY Exosomes mediate intercellular communication and their roles in a growing range of human disease is becoming increasingly evident. In preliminary studies, our lab has discovered that both virus- and LPS-induced exosome production is related to inflammasome activation resulting in cleavage of the Rab7 effector protein, RILP. The loss of RILP from the Rab7 complex reroutes intracellular trafficking toward the cell surface. This results in enhanced exosome secretion, due in part to a redistribution of multivesicular bodies throughout the cellular periphery. Formation of the cleaved form of RILP (cRILP) can also induce unique exosomal cargo loading, leading to the selective enrichment of specific pro-inflammatory miRNAs. Furthermore, inhibiting RILP cleavage can abrogate the proinflammatory actions generated by cRILP while specifically facilitating an anti-inflammatory response. This proposal will examine the hypothesis that enhanced exosome production that results from inflammatory disease is a consequence of inflammasome-triggered casepase-1 activation and subsequent cleavage of the trafficking adapter protein RILP. Cleaved RILP subsequently reprograms secretory events promoting stimuli-specific exosome formation and release. We will investigate this hypothesis with the following specific aims: Aim 1 will identify the mechanism by which cRILP regulates selective miRNA cargo loading. We hypothesize that the various forms of RILP may interact with and/or complex with specific subsets of RNA binding proteins that regulate miRNA cargo loading. Aim 2 will define the relationship between cRILP, RNABPs, and components of the ESCRT pathway. This aim will explore the hypothesis that cRILP hijacks the machinery required for exosome biogenesis leading to altered multivesicular body trafficking and subsequent exosome release. Aim 3 will examine the functional consequences of RILP manipulation on exosome secretion. This aim will identify how RILP manipulation, in both human monocytes and mouse models, can affect the outcome of various disease states to ultimately lead to the artificial modulation of exosome production and thus, the immune response. The ultimate goal of this research is to provide a detailed understanding of how exosomes are produced/secreted in response to pathogenic stimuli but also to describe the mechanisms that inflammatory stimuli use to specifically induce selective exosome cargo secretion. This knowledge can then be used to provide the tools to manipulate exosomes for therapeutic benefits.

项目摘要 外泌体介导细胞间通讯及其在越来越多的人类肿瘤中的作用 疾病变得越来越明显。在初步研究中，我们的实验室发现两种病毒- LPS诱导的外泌体产生与炎性小体激活有关，导致 Rab 7效应蛋白RILP。Rab 7复合物中RILP的丢失改变了细胞内的路线 向细胞表面运输。这导致增强的外泌体分泌，部分原因是 多泡体在整个细胞周边的重新分布。裂解形式的形成 RILP（cRILP）也可以诱导独特的外泌体货物装载，导致选择性富集 特定的促炎miRNAs。此外，抑制RILP切割可以消除RILP的活性。 cRILP产生的促炎作用，同时特异性促进抗炎作用， 反应这项提议将检验增强外泌体产生的假设， 炎症性疾病的结果是炎性小体触发的case 1 运输衔接蛋白RILP的活化和随后的切割。切开型RILP 随后重编程分泌事件，促进刺激特异性外泌体形成， release. 我们将研究这一假设与以下具体目标：目标1将确定 cRILP调节选择性miRNA货物装载的机制。我们假设 各种形式的RILP可以与RNA结合蛋白的特定亚类相互作用和/或复合 调节miRNA货物装载。目标2将定义cRILP，RNABP， 和ESCRT途径的组分。这一目标将探讨cRILP劫持了细胞的假设。 外泌体生物发生所需的机制，导致改变的多泡体运输， 随后外泌体释放。目标3将检查RILP的功能后果 操纵外泌体分泌。这一目标将确定RILP操纵，在人类和 单核细胞和小鼠模型，可以影响各种疾病状态的结果，最终导致 人工调节外泌体的产生，从而调节免疫反应。 这项研究的最终目标是提供一个详细的了解如何外泌体 产生/分泌的病原刺激，但也描述了机制， 用于特异性诱导选择性外泌体货物分泌的炎性刺激物。这些知识可以 然后用于提供操纵外泌体以获得治疗益处的工具。