Leukocyte subset identification by single cell analysis of BTB-ZF gene

通过 BTB-ZF 基因的单细胞分析鉴定白细胞亚群

• 批准号: 8705813
• 负责人: Derek B. Sant'Angelo
• 金额: $ 9.33万
• 依托单位: RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-15 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8705813
• 关键词: Antibodies; B cell differentiation; B-Lymphocytes; BCL6 gene; BTB/POZ Domain; Biological Assay; C-terminal; CD4 Positive T Lymphocytes; CD8B1 gene; Cell Lineage; Cell physiology; Cells; Coupled; Data; Development; Diagnostic tests; Effector Cell; Family; Family member; Flow Cytometry; Gene Expression; Gene Family; Genes; Helper-Inducer T-Lymphocyte; Human; Immune response; Immune system; Interleukin-17; Lead; Leukocytes; Lymphocyte; Lymphocyte Subset; Minor; Modeling; Mus; N-terminal; Pattern; Play; Population; Positioning Attribute; Protein Binding Domain; Publishing; Reagent; Regulatory Element; Regulatory T-Lymphocyte; Reporter; Role; Structure of germinal center of lymph node; System; T-Lymphocyte; T-Lymphocyte Subsets; Techniques; Transcriptional Regulation; Transgenes; Transgenic Mice; ZNF145 gene; Zinc Fingers; base; cell type; killer T cell; member; novel; novel diagnostics; pathogen; response; single cell analysis; thymocyte; transcription factor

DESCRIPTION (provided by applicant): Numerically minor subsets of lymphocytes have been found to have potent immunomodulatory functions. Indeed, the identification of T cell subsets such as regulatory T cells, T follicular helper cells, natural killer T cells and IL-17 producing T cell have, cumulatively, represented one of the most important advances in understanding the adaptive immune response. BTB-zinc finger (BTB-ZF) transcription factors are defined by an N-terminal protein-protein interaction domain (BTB/POZ) domain, coupled with a C-terminal zinc finger domain. Several members of this BTB-ZF family have emerged as fundamental, non-redundant factors that control the development or function of specific cell types of the immune system. For example, BTB-ZF genes have been shown to control T cell versus B cell commitment (LRF), CD4 versus CD8 lineage commitment (ThPOK), the commitment of thymocytes to innate T cell lineages (PLZF), the development of T follicular helper T cells (Bcl6) as well as differentiation of B cells into germinal center B cells (Bcl6). Additional members of ths gene family clearly influence the immune response, but in less well defined ways. In this R21 application we propose to explore BTB-ZF gene family expression to define novel leukocyte subset effector populations. We will develop a set of BAC-based transgenes that express both GFP and Cre under the control of the regulatory elements for BTB-ZF genes. Preliminary studies suggest that each of the selected BTB-ZF genes is expressed in the immune system, is regulated during development and, potentially, controls the function of a novel subset of cells. We are well positioned to carry out these studies and have already proven that the BAC reporter system is a highly effective approach. Indeed, we would strongly argue that this single cell expression model is essential for identifying leukocyte subsets and for the study of this important new family of transcription factors. The dramatic and nonredundant functions of the BTB-ZF genes that have thus far been identified clearly support the significance of the reagents we propose to develop and evaluate in the context of this R21 application. We believe that this effort will significantly advance the understanding of the transcriptional control of immune responses. Finally, since this family of transcription factors is highly conserved in humans, our studies have the potential to lead to new diagnostic tests and/or therapies.

描述（由申请方提供）：已发现淋巴细胞的数量较少的亚群具有强效免疫调节功能。事实上，T细胞亚群如调节性T细胞、T滤泡辅助细胞、自然杀伤T细胞和产生IL-17的T细胞的鉴定是非常重要的。 累积起来，代表了理解适应性免疫反应的最重要进展之一。BTB-锌指（BTB-ZF）转录因子由N-末端蛋白质-蛋白质相互作用结构域（BTB/POZ）结构域与C-末端锌指结构域偶联定义。这个BTB-ZF家族的几个成员已经成为控制免疫系统特定细胞类型的发育或功能的基本的非冗余因子。例如，BTB-ZF基因已显示控制T细胞相对于B细胞定型（LRF）、CD 4相对于CD 8谱系定型（ThPOK）、胸腺细胞向先天性T细胞谱系的定型（PLZF）、T滤泡辅助T细胞（Bcl 6）的发育以及B细胞向生发中心B细胞（Bcl 6）的分化。该基因家族的其他成员显然会影响免疫反应，但方式不太明确。在该R21申请中，我们提出探索BTB-ZF基因家族表达以定义新的白细胞亚群效应物群体。我们将开发一组基于BAC的转基因，其在BTB-ZF基因的调控元件的控制下同时表达GFP和Cre。初步研究表明，每个选定的BTB-ZF基因在免疫系统中表达，在发育过程中受到调节，并可能控制新细胞亚群的功能。我们有能力开展这些研究，并且已经证明BAC报告系统是一种非常有效的方法。事实上，我们强烈认为，这种单细胞表达模型是必不可少的识别白细胞亚群和研究这个重要的新家族的转录因子。到目前为止已经确定的BTB-ZF基因的显著和非冗余功能清楚地支持了我们提出在该R21应用的背景下开发和评估的试剂的意义。我们相信，这一努力将显着推进免疫反应的转录控制的理解。最后，由于这个转录因子家族在人类中高度保守，我们的研究有可能导致新的诊断测试和/或治疗。