Molecular Basis of cDC1 Development

cDC1 开发的分子基础

• 批准号: 10450553
• 负责人: Kenneth M Murphy
• 金额: $ 55.99万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-17 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10450553
• 关键词: ATAC-seq; Address; Antigen-Presenting Cells; Antigens; Binding; Bypass; CCAAT-Enhancer-Binding Proteins; CD8-Positive T-Lymphocytes; CRISPR/Cas technology; Cell physiology; Cell surface; Cells; ChIP-seq; Chimeric Proteins; Clinical; Complex; Critiques; Cross Presentation; Data; Dendritic Cells; Dependence; Development; Elements; Enhancers; Epitopes; Exhibits; FLT3 gene; FLT3 ligand; Family member; Gene Targeting; Generations; Genes; Genetic; Genetic Transcription; Goals; Homeostasis; Human; Immune response; Immunity; In Vitro; Infection; Literature; Lymphoid Tissue; MHC antigen; Methods; Mole the mammal; Molecular; Molecular Genetics; Mus; Myelogenous; Myeloid Cells; Output; Pathway interactions; Peptides; Population; Process; Production; Proteins; Publishing; Reagent; Reporter; Reporting; Role; Route; Signal Transduction; Site; Specific qualifier value; Suggestion; System; T cell response; T-Lymphocyte; Testing; Therapeutic; Time; Tumor-Derived; Vaccines; Viral; Virus; Work; adaptive immune response; design; effector T cell; granulocyte; in vivo; interest; macrophage; novel; progenitor; response; stem cells; tool; transcription factor; tumor; uptake

ABSTRACT The initial adaptive immune response to tumors and many viruses relies on the priming of CD8 T cells to gen- erate cytolytic effector T cells that can specifically target tumors or virally infected cells. The priming of CD8 T cells to these agents is carried out in vivo by a particular type of antigen presenting cell that is a component of the myeloid system and a member of the family of dendritic cells. Classical dendritic cells (cDCs) comprise several closely related lineages that are clearly distinct from other myeloid cells such as macrophages, mono- cytes or granulocytes. Primarily, cDCs serve to activate T cells against infections in the central lymphoid tis- sues, rather than carrying out direct effector functions at sites of infections as the other myeloid lineages do. The cDCs are themselves comprised of at least two major branches, now called cDC1 and cDC2. The cDC1 is a lineage that specializes in the uptake and processing of cell-associated antigens, such as from tumors of virally infected cells and the expression of peptide epitopes on its cell surface in conjunction with MHC-I mole- cules. This form of antigen:MHC-I complex is able to activate CD8 T cells, and not CD4 T cells. This process is called cross-presentation. The cDC2 is not capable of carrying out cross-presentation to viruses or tumors in vivo. The cDC1 has many genetic and molecular differences from cDC2; cDC1 require a distinct set of tran- scription factors for their development that are not required for cDC2. This includes dependence on the tran- scription factors Nfil3, Id2, Irf8 and Batf3. Our recent work showed that the genetic hierarchy among these fac- tors has Nfil3 as the first and initiating factor, acting to indirectly induce Id2 and Batf3 via the suppression of the repressor Zeb2. However, it is still unknown how Nfil3 is induced to initiate this process, and how Nfil3 works to suppress Zeb2 expression. It has recently become important to understand these details because of the clini- cal interest to apply Flt3L administration as a therapeutic in expanding the in vivo population of cDC1. It has been known for some time that Fl3L can expand dendritic cells in general and expand cDC1 in particular. But we have uncovered a surprising and worrisome fact; Flt3L administration will expand cDC1-like cells even in Nfil3-deficient mice, which completely lack cDC1 beforehand. The expansion of cDC1 in Nfil3-deficent mice produced by Flt3L is of the same magnitude as the expansion in WT mice. Thus, Flt3L is inducing cDC1 by a different genetic route than normal cDC1 development. There has been no test of whether such cDC1 cells function normally and will boost an immune response. This application will systematically address this issue by Aim 1) defining the normal process by which Nfil3 is induced, Aim 2) define the mechanism by which NFIL3 drives cDC1 development, and Aim 3) determine whether Flt3-induced cDC1 function normally and determine the mechanism by which Flt3L bypasses the normal requirement for Nfil3 in cDC1 development.

抽象的 对肿瘤和许多病毒的最初适应性免疫反应依赖于 CD8 T 细胞的启动 产生可特异性靶向肿瘤或病毒感染细胞的溶细胞效应 T 细胞。 CD8 T 的启动 细胞对这些试剂的作用是由特定类型的抗原呈递细胞在体内进行的，该细胞是 骨髓系统和树突状细胞家族的成员。经典树突状细胞 (cDC) 包括 几个密切相关的谱系，与其他骨髓细胞（如巨噬细胞、单核细胞）明显不同。 细胞或粒细胞。首先，cDC 的作用是激活 T 细胞以抵抗中央淋巴组织的感染。 起诉，而不是像其他骨髓谱系那样在感染部位执行直接效应功能。 cDC 本身至少由两个主要分支组成，现在称为 cDC1 和 cDC2。 cDC1 是专门从事细胞相关抗原的摄取和加工的谱系，例如来自肿瘤的抗原 病毒感染的细胞及其细胞表面肽表位的表达与 MHC-I 分子结合 库勒斯。这种形式的抗原：MHC-I 复合物能够激活 CD8 T 细胞，而不是 CD4 T 细胞。这个过程是 称为交叉呈现。 cDC2 不能与病毒或肿瘤进行交叉呈递 体内。 cDC1 与 cDC2 有许多遗传和分子差异； cDC1需要一组不同的反式 CDC2 不需要的其发育的转录因子。这包括对反式的依赖 转录因子 Nfil3、Id2、Irf8 和 Batf3。我们最近的工作表明，这些因素之间的遗传层次 tors 将 Nfil3 作为第一个启动因子，通过抑制 Id2 和 Batf3 来间接诱导 Id2 和 Batf3 阻遏物 Zeb2。然而，目前尚不清楚 Nfil3 是如何被诱导启动这一过程的，以及 Nfil3 是如何发挥作用的？ 抑制 Zeb2 表达。由于临床研究，了解这些细节最近变得很重要 人们有兴趣应用 Flt3L 给药作为扩大 cDC1 体内群体的治疗剂。它有 一段时间以来，人们知道Fl3L通常可以扩增树突状细胞，特别是可以扩增cDC1。但 我们发现了一个令人惊讶和令人担忧的事实； Flt3L 给药将扩增 cDC1 样细胞，即使在 Nfil3 缺陷小鼠，事先完全缺乏 cDC1。 Nfil3缺陷小鼠中cDC1的扩增 Flt3L 产生的量与 WT 小鼠中的扩增量相同。因此，Flt3L 通过 a 诱导 cDC1 与正常 CDC1 发育不同的遗传途径。还没有测试过这样的cDC1细胞是否 功能正常并会增强免疫反应。该应用程序将系统地解决这个问题 目标 1) 定义诱导 Nfil3 的正常过程，目标 2) 定义 NFIL3 的机制 驱动cDC1发育，目标3)确定Flt3诱导的cDC1功能是否正常并确定 Flt3L 绕过 CDC1 开发中对 Nfil3 的正常要求的机制。