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: 10407609
• 负责人: Declan McCole
• 金额: $ 52.33万
• 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/10407609
• 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; 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; gut inflammation; human model; improved; in vivo; 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 粘膜表面受到环境或微生物剂的挑战一直受到挑战。然而 粘膜屏障调节中先天免疫细胞之间相互作用的确切性质仍然是 理解不佳。该建议将结合一个具有多样和互补的调查员团队 在调节粘膜屏障中，研究先天免疫交叉言论机制的专业知识 功能。胃肠道衬里的单层上皮细胞是粘膜中最基本的元素 巨噬细胞在粘膜障碍稳态和对肠道的免疫力中起关键作用的障碍 病原体。这些肠上皮细胞（IEC）之间的交流的先前研究已重点放在 上皮功能的单向巨噬细胞调节。但是，我们认为这种观点过于 简单化，并且存在更复杂的监管电路。我们假设双向串扰 上皮细胞和巨噬细胞之间的电路在上皮的两个稳态调节中都起着关键作用 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谈话回路具有总体目标，以识别细胞内在如何 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， 反过来可以改变对方的功能能力。我们将在3个特定目标中检验我们的假设。 AIM 1将解决TCPTP如何调节单核细胞分化的连续性和巨噬细胞 粘膜稳态与肠道炎症的极化状态。 AIM 2将利用收养 转移实验以确定巨噬细胞中的TCPTP缺失与IEC如何调节肠道通透性 两种细胞类型在体内和体外的抗菌反应性。 AIM 3将识别分子 TCPTP调节这些细胞类型如何相互沟通的机制。我们有 为这项研究建立了新的小鼠系和体外共培养模型系统。我们将使用这些模型 一系列创新和既定的方法中的系统，使我们能够机械定义 这些基本相互作用的磷酸酶调节巨噬细胞和上皮细胞之间的这些基本相互作用 调节粘膜屏障功能和巨噬细胞极化。此外，我们将确定这些细胞是否是否 固有的磷酸酶调节的串扰机制通过使用小鼠和人类模型应用 系统。这些实验在基础上与先前的工作有关，但明确代表了令人兴奋的 新的指导使调查团队的专业知识协同。这些研究的结果是 有望在我们对先天基本基本机制的理解方面产生重大进步 粘膜表面的免疫生物学和细胞串扰。