Deciphering the immunoregulatory network governing antigen presenting myeloid cells

破译控制抗原呈递骨髓细胞的免疫调节网络

• 批准号: 10629283
• 负责人: Michael W Lipscomb
• 金额: $ 38.75万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629283
• 关键词: Antigen Presentation; Autoimmune Diseases; Biology; Calcium; Calcium Signaling; Cells; Complex; Coupled; Cyclic AMP-Dependent Protein Kinases; Dendritic Cells; Disease; Generations; Genes; Goals; Health; Hematopoiesis; Hematopoietic stem cells; Immune; Immune response; Immunity; Immunobiology; Immunologic Deficiency Syndromes; Immunotherapy; Infectious Agent; Inflammation; Knockout Mice; Knowledge; Laboratories; Macrophage; Malignant Neoplasms; Molecular; Myeloid Cells; Pathology; Pharmacotherapy; Physiological Processes; Play; Process; Protein Kinase C; Proteins; Publishing; RNA Interference; Research; Role; Signal Transduction; T cell response; Tissues; Work; allograft inflammatory factor-1; autoreactive T cell; combat; design; immunoregulation; in vivo; innovation; insight; insulitis; molecular mechanics; monocyte; novel; pathogen; phosphoric diester hydrolase; public health relevance; recruit; response; scaffold; screening; selective expression; success; tool; transcriptomic profiling

SUMMARY Dendritic cells (DC) are responsible for directing T cell responses and macrophages important for modulating tissue inflammation. A hallmark of these immune cells is their inherent plasticity to govern immunity vs. tolerance. However, much remains unknown regarding the complex molecular networks that collectively govern hematopoiesis, inflammation and antigen presentation. Incomplete understanding presents major obstacles to delineating their underlying roles in health and disease, which has also hindered success in developing effective immunotherapies. The long-term goal of the proposed research is to determine how pivotal immunoregulatory proteins govern DC and macrophage differentiation and immune responses. Published findings by my laboratory have begun to mechanistically define the functional roles of several important genes uniquely expressed in these myeloid cells. These genes, which include Allograft Inflammatory Factor-1 (AIF1) and Phosphodiesterase 1b (Pde1b), among others, were identified using a combination of high throughput transcriptomic profiling coupled with rigorous RNAi functional screening. My laboratory demonstrated that AIF1 is selectively expressed in conventional type 1 DC (cDC1), monocyte-derived DC (MoDC) and macrophages and serves as a scaffold to recruit protein kinase C (PKC) in a calcium-responsive manner to promote inflammation and type 1 polarized immune responses. Furthermore, AIF1 expression was required for successful generation of cDC1 from hematopoietic progenitors and both MoDC and macrophages from monocyte precursors. In the context of disease, AIF1 expression in DC and macrophages is required for initiating and sustaining insulitis and in regulating effector responses to intracellular pathogens. In another line of studies, the phosphodiesterase protein Pde1b was found to depress protein kinase A (PKA) activity in a calcium-dependent manner by regulating cyclonucleotide levels in cDC1, MoDC and macrophages to promote immune effector responses. Thus, my research group has begun to unravel how Pde1b works in concert with AIF1 to govern immunity by balancing PKC vs. PKA activities. As important as these initial findings are, there remains several gaps in understanding the molecular mechanics of how these genes govern immunobiology. As such, our research builds on prior studies by now employing conditional and global knockout mice and use of innovative experimental tools to rigorously study mechanistic roles in vivo. My laboratory will pursue the following major goals over the next five years: (1) delineate how AIF1 and Pde1B govern differentiation of DC and macrophages in vivo; (2) describe the intracellular processes by which other key novel immunoregulatory genes orchestrate immune responses; (3) determine the contributing role of AIF1 in initiating inflammation and sustaining autoreactive T cell responses; and (4) identify how intracellular pathogens antagonize AIF1 and Pde1b through disruption of calcium signaling and cyclonucleotide levels to evade host immunity. Knowledge gained will fill key gaps in our understanding of DC and macrophage biology and provide important insights into the molecular networks governing immunity.

概括 树突状细胞（DC）负责指导T细胞反应和巨噬细胞对调节很重要 组织炎症。这些免疫细胞的标志是控制免疫与公差的固有可塑性。 但是，关于集体控制的复杂分子网络仍然未知 造血，炎症和抗原表现。不完整的理解带来了主要障碍 描述其在健康和疾病中的潜在角色，这也阻碍了发展有效的成功 免疫疗法。拟议研究的长期目标是确定如何关键免疫调节 蛋白质控制DC和巨噬细胞分化和免疫反应。我的实验室发表的发现 已经开始机械地定义了在这些中唯一表达的几个重要基因的功能作用 髓样细胞。这些基因，包括同种异体移植炎性因子1（AIF1）和磷酸二酯酶1B （PDE1b）等使用高吞吐量转录组耦合的组合鉴定 具有严格的RNAi功能筛选。我的实验室证明AIF1在 常规1型DC（CDC1），单核细胞衍生的DC（MODC）和巨噬细胞，并用作脚手架 以钙响应方式募集蛋白激酶C（PKC），以促进炎症和1型极化 免疫反应。此外，成功从 造血祖细胞以及单核细胞前体的MODC和巨噬细胞。在 疾病，DC中的AIF1表达和巨噬细胞是启动和维持胰岛炎以及在 调节效应子对细胞内病原体的反应。在另一项研究中，磷酸二酯酶蛋白 发现PDE1b通过调节以钙依赖性方式降低蛋白激酶A（PKA）活性 CDC1，MODC和巨噬细胞中的氰核苷酸水平以促进免疫效应子反应。因此，我的 研究小组已开始解散PDE1B如何与AIF1协同工作以通过平衡来管理免疫力 PKC与PKA活动。与这些初步发现一样重要，理解仍然存在几个差距 这些基因如何控制免疫生物学的分子力学。因此，我们的研究基于先验 现在使用条件和全球敲除小鼠的研究以及使用创新的实验工具来进行研究 严格研究体内的机械作用。我的实验室将在接下来的五个中实现以下主要目标 年：（1）描述AIF1和PDE1B如何控制体内DC和巨噬细胞的分化； （2）描述 其他主要新型免疫调节基因协调免疫反应的细胞内过程； （3） 确定AIF1在启动炎症和维持自动反应性T细胞反应中的促进作用； （4）确定细胞内病原体如何通过破坏钙信号传导来拮抗AIF1和PDE1B 和环核苷酸水平以逃避宿主免疫。获得的知识将填补我们对 DC和巨噬细胞生物学，并提供有关控制免疫力的分子网络的重要见解。