Distinct functions of E protein family members in regulating γδ T lineage commitment and effector fate

E蛋白家族成员在调节γδT谱系承诺和效应子命运中的独特功能

• 批准号: 10226998
• 负责人: Juan Carlos Zuniga-Pflucker
• 金额: $ 30.97万
• 依托单位: RESEARCH INST OF FOX CHASE CAN CTR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10226998
• 关键词: Address; Adult; Antigens; Area; Autoimmune Diseases; Autoimmunity; Binding; Biological Models; CD34 gene; Cell Lineage; Cells; Chromatin; Clinical; Culture Techniques; Cytokine Receptors; DNA Binding; DNA-Binding Proteins; Data Set; Development; Disease Progression; E protein; EGR2 gene; Epigenetic Process; Epithelial Cells; Event; Exposure to; Family member; Flow Cytometry; Gene Expression Regulation; Genes; Genetic Transcription; Genomics; Goals; Health; Human; Immune system; In Vitro; Inflammation; Inflammatory; Interferons; Interleukin-17; Intervention; Joints; Knowledge; Lead; Link; Mediating; Mediator of activation protein; Molecular; Mucosal Immune System; Mucous Membrane; Mus; Mutant Strains Mice; Neonatal; Pathologic; Pattern; Peripheral; Play; Population; Population Heterogeneity; Process; Production; Proteins; Receptor Cell; Receptor Signaling; Regulator Genes; Role; Series; Signal Pathway; Signal Transduction; Site; Source; Specific qualifier value; Structure; T cell differentiation; T-Cell Development; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; TCF3 gene; Testing; Thymus Gland; Time; Tissues; Transcription Initiation Site; Translating; Tyrosine Phosphorylation; Umbilical Cord Blood; Work; cell fate specification; chromatin remodeling; combinatorial; cytokine; dimer; effector T cell; experimental study; fetal; genomic locus; granulocyte; human pluripotent stem cell; in vitro Model; in vivo Model; insight; member; mouse genetics; mouse model; mutant; notch protein; novel; novel marker; pathogen; preservation; progenitor; programs; protein function; recruit; response; single-cell RNA sequencing; transcription factor; transcriptome sequencing; tumor progression

PROJECT SUMMARY/ABSTRACT T cells belonging to the gd-lineage have been shown to serve unique and critical roles within the immune system. gd T cells are widely distributed throughout mucosal and epithelial cell-rich tissues and are an important early source of IL-17, which recruits granulocytes to sites of inflammation in response to pathogens. However, dysregulation of IL-17 production by gd T cells has also been linked to multiple pathological conditions including autoimmune disease and cancer progression. Unlike conventional ab T cells, gd T cells can acquire the ability to rapidly produce inflammatory cytokines within the thymus during development, prior to antigen exposure. The molecular and cellular events that specify and assign gd T cells to the IL-17 (gdT17) or interferon-g (IFNg; gdT1) effector fates during development remain to be fully elucidated. Recent evidence indicates that E proteins, in particular HEB, are critical mediators of this fate choice. HEB transcription factors are encoded by the Tcf12 gene locus, which gives rise to two proteins, HEBAlt and HEBCan, that differ in their structure, transcription initiation sites, and expression patterns. HEBAlt and HEBCan are expressed at partially overlapping times during gd T cell development, but their respective roles in functional programming of gd T cell precursors are unclear. We and others have identified a number of signaling pathways that contribute to gd T cell effector fate acquisition, including those that operate through T cell receptor (TCR), Notch, and cytokine receptors. It is our goal to understand how HEB activity is integrated with other determinants of gd T cell programming. Given our recent finding that HEB factors are required for the development of gdT17, but not gdT1 cells, we hypothesize that the final differentiation and effector function programming of gd T cells is directed by the interplay of specific HEB and E2A proteins with transcriptional regulators downstream of TCR, Notch, and cytokine receptor signaling. We will take advantage of a new in vitro model system that directs fate bifurcation between the gdT1 and gdT17 fates, as well as novel mouse models that we have generated, to elucidate the roles for E protein dimers containing HEBAlt, HEBCan, and/or E2A in determining gd T cell effector function, and to gain insight into the molecular basis for these events by assembling global gene regulatory networks. Our aims are: 1) to define the roles of HEBAlt, HEBCan, and E2A in intrathymic gd T cell differentiation by identifying specific target genes and binding partners in WT and mutant mice; and, 2) to evaluate the roles of HEB factors and their targets in human gd T cell development. Our proposal is built upon key observations derived from all Program Projects. The proposed experiments will continue to draw the complementary expertise in the areas of TCR signaling (Proj.1), mouse genetics (Proj.2), and chromatin remodeling (Proj.4). Our specific aims will not only lead to a better understanding of the mechanism of gd differentiation but also provide unique insights into the roles of specific E proteins in T cell development and function. Taken together, our joint experimental approaches and conceptual integration would not be possible without the combined expertise of all members of this Program.

项目摘要/摘要 属于gd血统的T细胞已被证明在免疫系统中起着独特和关键的作用。 GD T细胞广泛分布于粘膜和富含上皮细胞的组织中，是早期重要的 IL-17的来源，它招募粒细胞到炎症部位，以回应病原体。然而， Gd T细胞产生IL-17的失调也与多种病理情况有关，包括 自身免疫性疾病和癌症进展。与传统的ab T细胞不同，gd T细胞可以获得这种能力 在抗原暴露之前，在胸腺发育过程中迅速产生炎性细胞因子。这个 将gd T细胞指定和分配给IL-17(GdT17)或干扰素-g(ifg；gdT1)的分子和细胞事件 效应器在发育过程中的命运仍有待充分阐明。最近的证据表明，E蛋白，在 尤其是希伯来人，是这一命运选择的关键调解人。Heb转录因子由Tcf12编码 基因座，导致两种蛋白质HEBAlt和HEBCan的结构不同，转录 起始点和表达模式。HEBAlt和HEBCan在部分重叠的时间表达 Gd T细胞的发育，但它们各自在Gd T细胞前体功能编程中的作用尚不清楚。 我们和其他人已经确定了一些有助于获得gd T细胞效应器Fate的信号通路， 包括通过T细胞受体(TCR)、Notch和细胞因子受体发挥作用的受体。我们的目标是 了解Heb活性如何与gd T细胞编程的其他决定因素相结合。鉴于我们最近 发现Heb因子是gdT17细胞发育所必需的，而不是gdT1细胞所必需的，我们假设 GdT细胞的最终分化和效应功能编程是由特异性Heb的相互作用指导的 以及在TCR、Notch和细胞因子受体信号转导下游具有转录调节的E2a蛋白。我们 将利用一种新的体外模型系统来指导gdT1和gdT17命运之间的命运分歧， 以及我们建立的新的小鼠模型，以阐明E蛋白二聚体的作用，其中包括 HEBAlt、HEBCan和/或E2a在确定gd T细胞效应器功能和深入了解分子方面的作用 通过组建全球基因调控网络，为这些活动奠定基础。我们的目标是：1)确定 HEBAlt、HEBCan和E2a与胸腺内gd T细胞分化的关系 以及，2)评估Heb因子及其靶点在WT和突变小鼠中的作用。 人GdT细胞发育。我们的建议建立在从所有计划项目中得出的关键观察结果的基础上。 拟议的实验将继续吸收TCR信令领域的补充专业知识 (项目1)、小鼠遗传学(项目2)和染色质重塑(项目4)。我们的具体目标不仅将导致 更好地理解gd分化的机制，也提供了对gd分化的作用的独特见解。 T细胞发育和功能中的特异性E蛋白。综上所述，我们的联合实验方法和 如果没有本计划所有成员的综合专业知识，概念整合是不可能的。