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.

项目概要 外泌体介导细胞间通讯及其在越来越多的人类中的作用 疾病越来越明显。在初步研究中，我们的实验室发现这两种病毒- LPS诱导的外泌体产生与炎症小体激活相关，导致裂解 Rab7 效应蛋白 RILP。 Rab7 复合体中 RILP 的丢失改变了细胞内的路线 向细胞表面运输。这导致外泌体分泌增强，部分原因是 多泡体在整个细胞外周的重新分布。裂解形式的形成 RILP (cRILP) 还可以诱导独特的外泌体货物装载，从而导致选择性富集 特定的促炎 miRNA。此外，抑制 RILP 裂解可以消除 cRILP 产生的促炎作用，同时特异性促进抗炎作用 回复。该提案将检验以下假设：增强外泌体的产生 炎症性疾病的结果是炎症小体触发的 casepase-1 的结果 运输接头蛋白 RILP 的激活和随后的裂解。裂开的RILP 随后重新编程促进刺激特异性外泌体形成的分泌事件 发布。 我们将通过以下具体目标来研究这一假设： 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 可能与 RNA 结合蛋白的特定子集相互作用和/或复合 调节 miRNA 货物装载。目标 2 将定义 cRILP、RNABP、 以及 ESRT 途径的组成部分。该目标将探讨 cRILP 劫持的假设 外泌体生物发生所需的机制导致改变多泡体运输和 随后的外泌体释放。目标 3 将检查 RILP 的功能后果 外泌体分泌的调控。该目标将确定 RILP 在人类和人类中的操纵方式 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详细了解外泌体是如何发挥作用的。 产生/分泌以响应致病刺激，但也描述了机制 炎症刺激用于特异性诱导选择性外泌体分泌。这些知识可以 然后用于提供操纵外泌体以获得治疗效果的工具。