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 外泌体介导细胞间交流及其在不断增长的人类中的作用 疾病越来越明显。在初步研究中，我们的实验室发现两种病毒 - LPS诱导的外泌体产生与炎性体激活有关，导致裂解 Rab7效应蛋白，RILP。 RAB7复合物中的RILP损失细胞内重新布置 运输向细胞表面。这导致了增强的外泌体分泌，部分原因是 整个细胞周围的多囊体重新分布。裂片形式的形成 RILP（CRILP）也可以引起独特的外泌体货物载荷，从而导致选择性富集 特定的促炎性miRNA。此外，抑制RILP裂解可以消除 CRILP产生的促炎作用，同时特别促进抗炎 回复。该提案将研究以下假设，即增强了外泌体生产 炎症性疾病的结果是炎性体触发的casepase-1的结果 运输适配器蛋白RILP的激活和随后的切割。切割的rilp 随后重新编程的分泌事件促进了特定刺激的外泌体形成和 发布。 我们将以以下特定目的调查这一假设：AIM 1将确定 CRILP调节选择性miRNA货物载荷的机制。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/或复合物 调节miRNA货物载荷。 AIM 2将定义CRILP，rnabps之间的关系 和ESCRT途径的组成部分。这个目的将探讨以下假设： 外泌体生物发生所需的机械导致多孔体贩运和 随后的外泌体释放。 AIM 3将检查RILP的功能后果 对外泌体分泌的操纵。这个目标将确定在人类中如何操纵RILP 单核细胞和小鼠模型会影响各种疾病状态的结果，最终导致 外泌体产生的人工调制，因此是免疫反应。 这项研究的最终目的是提供对外泌体的详细了解 响应致病性刺激而产生/分泌 炎症刺激用于特异性诱导选择性外泌体货物分泌。这些知识可以 然后被用来提供操纵外泌体的工具以获得治疗益处。