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: 10397059
• 负责人: Ann Lynn Wozniak
• 金额: $ 42.67万
• 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 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10397059
• 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; Event; Goals; Guanosine Triphosphate Phosphohydrolases; Human; Immune response; In Vitro; Inflammasome; Inflammatory; Inflammatory Response; Kinesin; Knowledge; Lead; Link; Liver; Liver diseases; Measures; Mediating; Membrane; MicroRNAs; Microtubules; Modification; Molecular Motors; Movement; Multivesicular Body; Outcome; Parents; Pathogenicity; Pathologic; Pathologic Processes; Pathway interactions; Patients; Physiological; Physiological Processes; Play; Process; Production; Proteins; Proteolysis; RNA; RNA-Binding Proteins; Recruitment Activity; Regulation; Research; Role; Site; Sorting - Cell Movement; Specificity; Stimulus; Testing; Therapeutic; Tissue Sample; Ubiquitination; Vesicle; Virus; Work; cell type; exosome; human disease; in vivo; intercellular communication; lysosomal proteins; macrophage; monocyte; mouse model; nano-exosomes; nanosized; novel; patient population; protein protein interaction; rab7 protein; response; tool; trafficking; vesicular release; 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.

项目总结 外切体介导胞间通讯及其在越来越多的人类中的作用 疾病正变得越来越明显。在初步研究中，我们的实验室发现这两种病毒- 而内毒素诱导的外切体的产生与炎性小体的激活有关，从而导致裂解 Rab7效应蛋白，RILP。Rab7复合体细胞内重路由中RILP的丢失 向细胞表面运输。这导致外切体分泌增加，部分原因是 多泡小体在细胞周围重新分布。卵裂型的形成 RILP(CRILP)还可以诱导独特的胞外货物装载，从而导致选择性浓缩 特定的促炎miRNAs。此外，抑制RILP的切割可以消除 由cRILP产生的促炎作用，同时特别促进抗炎 回应。这项提议将检验这样一种假设，即增强外切体的产生 炎症性疾病的结果是炎症组触发的Casepase-1的结果 运输适配蛋白RILP的激活和随后的切割。劈裂RILP 随后对促进刺激特异性外切体形成的分泌事件进行重新编程 放手。 我们将以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可以与特定的RNA结合蛋白亚群相互作用和/或与其形成复合体 来调节miRNA货物的装载。目标2将定义cRILP、RNABP、 和ESCRT途径的组成部分。这一目标将探索cRILP劫持 外体生物发生所需的机制导致改变的多囊体体运输和 随后的外切体释放。目标3将检查RILP的功能后果 对外切体分泌的操纵。这一目标将确定RILP操作如何在人类和 单核细胞和小鼠模型，可以影响各种疾病状态的结局，最终导致 人工调节外切体的产生，从而调节免疫反应。 这项研究的最终目标是详细了解外切体是如何 对致病刺激的反应而产生/分泌，也描述了 炎性刺激用于特异性地诱导选择性外切体货物的分泌。这一知识可以 然后被用来提供工具来操纵外显体以获得治疗效果。