Role of nuclear factor TOX in lymphocyte development

核因子 TOX 在淋巴细胞发育中的作用

基本信息

批准号：
9751160
负责人：
JONATHAN G KAYE
金额：
$ 43.75万
依托单位：
CEDARS-SINAI MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-12-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9751160
关键词：
Address Area Binding Binding Proteins Biological Models Bone Marrow CD4 Positive T Lymphocytes Cell Differentiation process Cell Fate Control Cell Lineage Cells ChIP-seq Chromatin Remodeling Factor Collection Common Lymphoid Progenitor Complex DNA-Binding Proteins Data Development Disease Epitopes Event Gene Expression Gene Proteins Gene Targeting Genes Genetic Transcription HBXAP gene HMG Domain HMG-Box HMGB Proteins IL2RA gene ISWI Immune Immune system Immunity Immunoprecipitation In Vitro Innate Immune System Kinetics Knockout Mice Knowledge Lead Lymphocyte Lymphocyte Subset Lymphoid Cell Mass Spectrum Analysis Mediating Modeling Molecular Mouse Strains Mus Natural Killer Cells Nuclear Pattern Play Population Process Proteins Regulation Reporter Research Project Grants Role SMARCA5 gene Signal Transduction Stem cells System T-Cell Development T-Lymphocyte Testing Thymocyte Selection Thymus Gland Tissue-Specific Gene Expression Up-Regulation Work cell type in vitro Model in vivo interest member notch protein novel precursor cell progenitor single cell analysis thymocyte tool transcriptome

项目摘要

Project Summary/Abstract Only recently has the extent of the diversity of innate immune effector functions become apparent, with the discovery of multiple novel subtypes of innate lymphoid cells (ILCs), beyond long recognized natural killer cells. ILCs derive from common lymphoid progenitors (CLP) in the bone marrow, and a number of transcriptional regulators that are involved in ILC development also play roles in the development of T cells in the thymus. Among these is TOX (thymocyte-selection associated HMG-box protein), a nuclear DNA binding protein and member of the HMG-box superfamily of proteins. Our previous work has shown that TOX is required for CD4 T cell lineage development in the thymus, and for early ILC lineage specification. Using a TOX reporter strain of mouse we have now identified bone marrow progenitor cells that may be in transition from CLP to the common progenitor to all helper-like innate lymphoid cells (CHILP). CHILP highly express the transcriptional regulator Id2, but we have identified Tox+Id2lo putative precursor cells, suggesting that upregulation of TOX likely precedes the formation of CHILP. We propose to test the cell fate potential of these cells in vitro and in vivo, to determine what ILC lineages they can form and if they have lost T cell potential. Similarly, we will ask if ILC lineage specification even precedes Tox upregulation. Armed with this knowledge, we then propose to determine the transcriptome in these cell types, at the single cell level, and to determine the influence of TOX on gene expression. Notch signaling is also thought to play a role in early ILC lineage specification. We propose that transient Notch signaling is key to distinguish ILC from T cell lineage commitment. Using an in vitro model system that allows differentiation of CLP to multiple ILC subtypes, we will study the cell fate and transcriptional consequences of a temporally limited Notch signal. As Tcf7 (encoding TCF-1), a key Notch signaling target gene, is likely downstream of TOX, we ask if expression of Tcf7 can compensate for loss of TOX. Finally, we address the underlying mechanism of action of TOX, using in vitro and in vivo approaches to ask if TOX functions as part of a novel RSF ISWI chromatin-remodeling complex, and propose to produce a novel mouse strain to aid in identification of gene targets and protein binding partners of TOX in multiple cell contexts.

项目摘要/摘要直到最近才使先天免疫效应子功能的多样性变得显而易见，并且发现了先天淋巴样细胞（ILC）的多种新型亚型，超出了长期公认的天然杀手细胞。 ILC源自骨髓中常见的淋巴祖细胞（CLP），许多参与ILC开发的转录调节剂也在T细胞的发展中起作用在胸腺中。其中包括毒物（胸腺丝体相关的HMG-box蛋白），一种核DNA 结合蛋白和蛋白质HMG-box超家族的成员。我们以前的工作表明，胸腺中CD4 T细胞谱系的发育需要托克斯，以及早期的ILC谱系规范。使用小鼠的托克斯报道菌株我们现在已经鉴定出骨头骨髓祖细胞可能从CLP过渡到公共祖细胞到所有辅助型先天性淋巴样细胞（CHILP）。高度表达转录调节剂ID2，但我们已经确定了 TOX+ID2LO推定的前体细胞，表明TOX的上调可能是在形成之前辣椒。我们建议在体外和体内测试这些细胞的细胞命运潜力，以确定什么ILC 它们可以形成谱系，如果它们失去了T细胞电位。同样，我们会询问ILC血统是否规格甚至先于托克斯上调。凭借这些知识，我们建议确定这些细胞类型的转录组，在单细胞水平上，并确定毒物对基因的影响表达。 Notch信号传导还被认为在早期ILC谱系规范中起作用。我们建议该瞬态Notch信号传导是区分ILC和T细胞谱系承诺的关键。使用体外允许将CLP分化为多个ILC亚型的模型系统，我们将研究细胞命运和时间限制的Notch信号的转录后果。作为TCF7（编码TCF-1），键缺口信号靶基因可能是托克斯的下游，我们询问TCF7的表达是否可以补偿毒品。最后，我们使用体外和体内方法解决了托克斯的基本作用机理询问托克斯是否作为新型RSF ISWI染色质复制复合物的一部分，并提议产生一种新的小鼠菌株，以帮助鉴定托克斯的基因靶标和蛋白质结合伴侣多个单元环境。