Role of nuclear factor TOX in lymphocyte development

项目总结/摘要 直到最近，先天免疫效应子功能的多样性程度才变得明显， 发现了多种新的先天淋巴样细胞（ILC）亚型， 杀伤细胞ILC来源于骨髓中的共同淋巴样祖细胞（CLP），并且许多ILC是由细胞因子和细胞因子共同作用的。 参与ILC发育的转录调节因子也在T细胞的发育中发挥作用 在胸腺中。其中之一是TOX（胸腺细胞选择相关HMG盒蛋白），一种核DNA 结合蛋白和HMG盒蛋白超家族成员。 我们以前的工作表明，TOX是胸腺中CD 4 T细胞谱系发育所必需的， 以及早期ILC谱系规范。使用TOX报告小鼠品系，我们现在已经确定了骨 骨髓祖细胞，可能是在过渡，从CLP的共同祖细胞，以所有辅助样先天 淋巴样细胞（CHILP）。CHILP高度表达转录调节因子Id 2，但我们已经鉴定出， Tox+ Id 2lo推定的前体细胞，这表明TOX的上调可能先于Id 2lo的形成。 冷静。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ILC 他们可以形成的谱系，如果他们已经失去了T细胞的潜力。同样，我们会问，如果ILC血统 说明甚至先于Tox上调。有了这些知识，我们建议确定 在这些细胞类型中，在单细胞水平上的转录组，并确定TOX对基因表达的影响。 表情Notch信号传导也被认为在早期ILC谱系特化中起作用。我们提出 瞬时Notch信号是区分ILC和T细胞谱系定型的关键。使用体外 模型系统，允许分化的CLP多种ILC亚型，我们将研究细胞的命运， 时间有限的Notch信号的转录后果。作为Tcf 7（编码TCF-1）， 信号转导靶基因，可能是TOX的下游，我们想知道Tcf 7的表达是否可以弥补TOX的损失。 TOX.最后，我们使用体外和体内的方法来解决TOX的潜在作用机制 询问TOX是否作为新型RSF ISWI染色质重塑复合物的一部分发挥作用，并提出 产生一种新的小鼠品系，以帮助鉴定基因靶点和TOX的蛋白质结合伴侣， 多个细胞上下文。

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.