Tyrosine Phosphatase Regulation of Mucosal Macrophage-Epithelial Cell Cross-talk

酪氨酸磷酸酶对粘膜巨噬细胞-上皮细胞串扰的调节

• 批准号: 10031958
• 负责人: Declan McCole
• 金额: $ 51.24万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10031958
• 关键词: Activities of Daily Living; Address; Adoptive Transfer; Affect; Anti-Inflammatory Agents; Biological Models; Biology; Cell Communication; Cell Line; Cell physiology; Cells; Citrobacter rodentium; Coculture Techniques; Communication; Complex; DNA Sequence Alteration; Data; Elements; Epithelial; Epithelial Cells; Epithelium; Event; Exposure to; Foundations; Gastrointestinal tract structure; Goals; Homeostasis; Host Defense; Human; Immune; Immune response; Immunity; Immunobiology; In Vitro; Infection; Inflammation; Intestinal permeability; Intestines; Knock-out; Knockout Mice; Link; Maintenance; Mediating; Mediator of activation protein; Modification; Molecular; Mucous Membrane; Mus; Myelogenous; Nature; Peptide Signal Sequences; Permeability; Phosphoric Monoester Hydrolases; Play; Property; Protein Tyrosine Phosphatase; Regulation; Research Personnel; Role; Series; Signal Pathway; Signal Transduction; Surface; System; T-cell protein tyrosine phosphatase; Testing; Tight Junctions; Transgenic Mice; Work; antimicrobial; behavior change; cell growth regulation; cell type; chronic autoimmune disease; cytokine; enteric pathogen; experimental study; human model; improved; in vivo; inflammatory disease of the intestine; innate immune function; innate immune mechanisms; innovation; intestinal barrier; intestinal epithelium; intestinal homeostasis; macrophage; microbial; monocyte; mouse model; novel; preservation; response

SUMMARY/ABSTRACT Mucosal surfaces are under constant challenge from exposure to environmental or microbial agents. However the exact nature of interactions between innate immune cells in the regulation of the mucosal barrier is still poorly understood. This proposal will incorporate a team of investigators with diverse and complementary expertise to investigate the mechanisms of innate immune cross-talk in the regulation of mucosal barrier function. The single layer of epithelial cells lining the GI tract is the most fundamental element of the mucosal barrier while macrophages play a key role in mucosal barrier homeostasis and in immunity to intestinal pathogens. Previous studies of communication between these intestinal epithelial cells (IEC) have focused on unidirectional macrophage regulation of epithelial function. However, we believe that this view is overly simplistic and that a more complex regulatory circuit exists. We hypothesize that bidirectional cross-talk circuits between epithelial cells and macrophages play a key role in both homeostatic regulation of epithelial permeability and macrophage polarization. Here, we will investigate the molecular nature of bidirectional cross- talk circuits between epithelial cells and macrophages with the overall objective to identify how the cell-intrinsic activity of an essential protein tyrosine phosphatase (TCPTP) regulates molecular changes in one cell type, which can in turn modify the functional capacity of the other. We will test our hypothesis in 3 Specific Aims. Aim 1 will address how TCPTP modulates the monocyte differentiation continuum and macrophage polarization status in mucosal homeostasis vs. local inflammation in the intestine. Aim 2 will utilize adoptive transfer experiments to identify how TCPTP deletion in macrophages vs. IEC modulates intestinal permeability and antimicrobial responsiveness of both cell types in vivo and in vitro. Aim 3 will identify molecular mechanisms by which TCPTP regulates how these cell types cross-communicate with each other. We have established novel mouse lines and in vitro co-culture model systems for this study. We will use these model systems in a series of innovative and established approaches, to allow us to mechanistically define phosphatase regulation of these fundamental interactions between macrophages and epithelial cells in the regulation of mucosal barrier function and macrophage polarization. In addition, we will identify if these cell- intrinsic phosphatase regulated crosstalk mechanisms apply across species by using mouse and human model systems. These experiments are foundationally linked to prior work but unequivocally represent an exciting new direction that synergizes the expertise of the investigative team. The results from these studies are poised to generate significant advances in our understanding of fundamental basic mechanisms in innate immunobiology and cellular crosstalk at mucosal surfaces.

摘要/摘要 粘膜表面因暴露于环境或微生物而不断受到挑战。然而 先天免疫细胞之间在粘膜屏障调节中相互作用的确切性质仍不清楚 不太了解。该提案将包含一个由多元化和互补性的研究人员组成的团队 研究先天免疫串扰调节粘膜屏障的机制的专业知识 功能。胃肠道内衬的单层上皮细胞是粘膜最基本的组成部分 屏障，而巨噬细胞在粘膜屏障稳态和肠道免疫中发挥关键作用 病原体。先前对这些肠上皮细胞（IEC）之间通讯的研究主要集中在 巨噬细胞对上皮功能的单向调节。但我们认为这种观点有些过分 简单化，并且存在更复杂的调节电路。我们假设双向串扰 上皮细胞和巨噬细胞之间的回路在上皮细胞稳态调节中发挥着关键作用 渗透性和巨噬细胞极化。在这里，我们将研究双向交叉的分子性质 上皮细胞和巨噬细胞之间的对话回路，总体目标是确定细胞内在的 必需蛋白酪氨酸磷酸酶（TCPTP）的活性调节一种细胞类型的分子变化， 这反过来又可以改变对方的功能能力。我们将在 3 个具体目标中检验我们的假设。 目标 1 将解决 TCPTP 如何调节单核细胞分化连续体和巨噬细胞 粘膜稳态的极化状态与肠道局部炎症的比较。目标 2 将利用收养 转移实验以确定巨噬细胞中 TCPTP 缺失与 IEC 如何调节肠道通透性 以及体内和体外两种细胞类型的抗菌反应性。目标 3 将识别分子 TCPTP 调节这些细胞类型如何相互交叉通讯的机制。我们有 为此研究建立了新的小鼠品系和体外共培养模型系统。我们将使用这些模型 系统采用一系列创新和既定的方法，使我们能够机械地定义 磷酸酶对巨噬细胞和上皮细胞之间这些基本相互作用的调节 粘膜屏障功能和巨噬细胞极化的调节。此外，我们将确定这些细胞是否 通过使用小鼠和人类模型，内在磷酸酶调节的串扰机制适用于跨物种 系统。这些实验基本上与之前的工作相关，但明确代表了令人兴奋的结果 协同调查团队专业知识的新方向。这些研究的结果是 有望在我们对先天基本机制的理解方面取得重大进展 粘膜表面的免疫生物学和细胞串扰。