Microbial Control of Host Intercellular Communication

宿主细胞间通讯的微生物控制

基本信息

批准号：
10363966
负责人：
REBECCA L LAMASON
金额：
$ 30.5万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-15 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10363966
关键词：
Actins Adhesions Bacteria Biological Biology Buffers CAV1 gene CAV2 gene Cardiovascular Diseases Caveolae Caveolins Cell physiology Cell-Cell Adhesion Cells Cellular biology Communication Communications Media Complex Cytoskeleton Development Disease Endocytosis Eukaryotic Cell Genes Goals Health Homeostasis Human Image Immunology Intercellular Junctions Life Listeria Listeria monocytogenes Malignant Neoplasms Mechanical Stress Mechanics Mediating Membrane Microbe Microscopy Molecular Molecular Probes Organelles Organism Pathway interactions Polymers Process Protein Dynamics Proteins Quantitative Microscopy RNA Interference RNA interference screen Regulation Role Shapes Signal Transduction Small Interfering RNA Structure Tail Testing Tissues Work base cell motility host-microbe interactions human disease human tissue improved innovation insight intercellular communication knock-down mechanical signal mechanotransduction microbial novel pathogenic bacteria recruit response spatiotemporal tool

项目摘要

PROJECT SUMMARY/ABSTRACT Multicellular organisms use intercellular communication to coordinate cell function and maintain tissue homeostasis. Recent work suggests that this communication is driven in part by the exchange of organelles (via trans-endocytosis) and mechanical cues directly across cell-cell junctions. Dysregulation of this communication leads to cancer and cardiovascular diseases. Despite its importance, we lack a fundamental molecular understanding of how intercellular communication occurs because of the limited number of cell biological tools capable of probing the molecular mechanisms at cell-cell contacts. This proposal seeks to elucidate the regulatory mechanisms of the pathways thought to control intercellular communication by studying how they are manipulated when under microbial control. The bacterium Listeria monocytogenes disseminates through human tissues using a process called cell-to-cell spread, which is a vesicular-mediated form of intercellular exchange that mimics host trans-endocytosis. Listeria spreads from cell to cell by mobilizing the host’s actin cytoskeleton for intracellular motility and transport to the cell-cell junction. Once at the junction, it pushes against the membrane and forms a double-membrane protrusion that is engulfed by a neighboring cell. Studying this distinctive spreading process will allow us to examine several outstanding cell biological questions. First, are specific endocytic pathways used at cell-cell junctions to engulf large cargo like microbes? Second, are mechanically-sensitive membrane domains or membrane curvature proteins activated as Listeria pushes against the junction during spread? To answer these questions, we used a high-content, image-based siRNA screen to test if Listeria requires host intercellular communication pathways during spread. We discovered that the endocytic and mechanoresponsive caveolar proteins CAV1, CAV2, and PACSIN2 promote Listeria spread. We also revealed a putative role for 19 other host proteins, including those that regulate membrane curvature, trans- endocytosis, and adhesion. Our preliminary findings suggest the overall hypothesis that Listeria subverts multiple intercellular communication pathways to promote cell-to-cell spread. In Aim 1, we will determine how PACSIN2 and caveolins coordinate their activities to promote the engulfment stage of cell-to-cell spread. In Aim 2, we will reveal which of the remaining hits regulate Listeria spread specifically, how they function, and if they work independently or together with caveolae. In the end, our proposed studies will improve our fundamental understanding of host-microbe interactions and basic cell biology, and may uncover how intercellular communication goes awry in human disease.

项目概要/摘要多细胞生物利用细胞间通讯来协调细胞功能和维持组织最近的研究表明，这种交流部分是由细胞器的交换驱动的。反式内吞作用）和直接跨细胞-细胞连接的机械线索。尽管它很重要，但我们缺乏一种基本的分子。由于细胞生物学工具数量有限，了解细胞间通讯如何发生该提案旨在阐明细胞与细胞接触的分子机制。通过研究细胞间通讯的方式来控制细胞间通讯的途径的调节机制在微生物控制下进行操纵，单核细胞增生李斯特氏菌通过人体传播。组织使用一种称为细胞间传播的过程，这是一种囊泡介导的细胞间交换形式模仿宿主的反内吞作用，李斯特菌通过动员宿主的肌动蛋白细胞骨架在细胞之间传播。用于细胞内运动和运输到细胞与细胞连接处。膜并形成双膜突出物，被邻近的细胞吞噬。独特的传播过程将使我们能够研究几个突出的细胞生物学问题。细胞与细胞连接处使用的特定内吞途径来吞噬微生物等大型货物？当李斯特菌推动时，机械敏感膜域或膜曲率蛋白被激活为了回答这些问题，我们使用了高内涵、基于图像的 siRNA 筛选来测试李斯特菌在传播过程中是否需要宿主细胞间通讯途径。内吞性和机械反应性小凹蛋白 CAV1、CAV2 和 PACSIN2 促进李斯特菌传播。还揭示了 19 种其他宿主蛋白的假定作用，包括那些调节膜曲率、反式我们的初步研究结果表明李斯特菌颠覆了多种作用的总体假设。细胞间通讯途径促进细胞间传播在目标 1 中，我们将确定 PACSIN2 如何发挥作用。在目标 2 中，我们将协调它们的活动以促进细胞间传播的吞噬阶段。揭示哪些剩余的热门产品专门调节李斯特菌的传播，它们如何发挥作用，以及它们是否有效最后，我们提出的研究将改善我们的基础。了解宿主-微生物相互作用和基础细胞生物学，并可能揭示细胞间相互作用的机制人类疾病中沟通出现问题。