Study Sox4 regulated genes in B cell development in a novel in vitro system

在新型体外系统中研究 B 细胞发育中的 Sox4 调节基因

• 批准号: 8638280
• 负责人: XIAOPING SUN
• 金额: $ 24万
• 依托单位: UNIVERSITY OF TX MD ANDERSON 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: 2014-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8638280
• 关键词: Adult; Award; B-Cell Development; B-Lymphocytes; Binding Sites; Cardiac; Cell Culture Techniques; Cell Lineage; Cell Survival; ChIP-seq; Common Lymphoid Progenitor; Cultured Cells; Data; Defect; Development; Developmental Gene; Developmental Process; Disease; Down-Regulation; Embryo; Event; Fetal Liver; Future; Gene Chips; Genes; Genetic Transcription; Genome; Hematopoietic stem cells; Hepatocyte; Humoral Immunities; Immune System Part; Immune system; Immunologic Deficiency Syndromes; Immunology; In Vitro; Investigation; Journals; Knock-out; Knockout Mice; Knowledge; Label; Mature B-Lymphocyte; Modeling; Molecular; Mouse Strains; Mus; National Institute of Allergy and Infectious Disease; Play; Population; Process; Publications; Publishing; Rag1 Mouse; Regulation; Reporting; Research; Role; Staging; System; T-Lymphocyte; Testing; Time; Transplantation; Work; congenital immunodeficiency; humoral immunity deficiency; in vivo; interest; mouse model; novel; progenitor; programs; public health relevance; research study; transcription factor

SunR21 Project Summary/Abstract B cells constitute an integral part of the immune system. B cell deficiency is one of the major causes of primary immunodeficiency diseases. Of more than 120 known PIDs, B cell deficiencies and combined B and T cell deficiencies are responsible for nearly 40 of them. B cell development is a sequential process consisting of the following stages: hematopoietic stem cell, multilineage progenitor, common lymphoid progenitor, pre-pro-B (fraction A), pro-B (fractions B and C), early pre-B (fraction C'), late pre-B (fraction D), immature B (fraction E), and mature B (fraction F). The process is precisely regulated by coordinated functioning of multiple genes, especially those encoding transcription factors. Sox4 is a gene that encodes a transcription factor and is known to be vital for many developmental processes. Sox4 conventional knockout causes embryos to die due to cardiac deficiency. Fetal liver cells isolated from Sox4 knockout mice fail to generate B cells in recipient mice after transplantation, indicating that the function of Sox4 is indispensable for B cell development. The lethality of the Sox4 conventional knockout has hindered further studies on the role of Sox4 in B cell development. We systematically investigated the role of Sox4 in B cell development using conditional knockout mouse models with Sox4 deletion at different developmental stages. Sox4 deletion in hematopoietic stem cells results in a nearly complete lack of pro-B cells and later stage B cells, whereas Sox4 deletion in pre-B cells or mature B cells has no detectable effect. Although the conditional knockout models have allowed us to reveal the developmental stage-specific functions of Sox4, the lack of B cells has imposed an obstacle for future in-depth studies. To overcome this problem, we have developed an in vitro culture system in which a stage specific developmental event is dependent on the presence of Sox4. This functional system is ideal for characterizing the molecular mechanisms underlying Sox4 function. We have performed gene expression microarray profiling using cultured cells and identified potential Sox4-regulated genes. To pursue in-depth characterization of Sox4 function in B cell development using our novel in vitro system, we propose two specific aims for this project. Aim 1 is to identify and characterize genes directly regulated by Sox4. Aim 2 is to characterize the roles of two downstream targets of Sox4 in early B cell development. Successful completion of this proposed research will elucidate the transcription program regulated by Sox4, advance our understanding of B cell development, and expand our knowledge of PIDs.

SunR21项目摘要/摘要 B细胞是免疫系统不可或缺的一部分。B细胞缺乏是原发疾病的主要原因之一。 免疫缺陷疾病。在120多个已知的ID中，B细胞缺陷和B细胞和T细胞联合 其中近40个是由缺陷造成的。B细胞的发育是一个连续的过程，包括 以下阶段：造血干细胞，多系祖细胞，共同淋巴祖细胞，前前B (组分A)、前-B组分(组分B和C)、早期前B组分(组分C‘)、晚期前B组分(组分D)、未成熟B组分(组分E)、 和成熟的B(组分F)。这一过程受到多个基因协同作用的精确调控， 尤其是那些编码转录因子的基因。Sox4是一种编码转录因子的基因，已知 对许多发育过程至关重要。Sox4常规基因敲除导致胚胎死亡 心脏缺乏症。从Sox4基因敲除小鼠分离的胎肝细胞不能在受体小鼠中产生B细胞 移植后，提示Sox4的功能对B细胞的发育是不可或缺的。杀伤力 Sox4基因的常规敲除阻碍了对Sox4在B细胞发育中作用的进一步研究。我们 利用条件性基因敲除小鼠模型系统研究了Sox4在B细胞发育中的作用 不同发育阶段的Sox4缺失。造血干细胞中SOX4缺失导致 几乎完全缺乏前B细胞和晚期B细胞，而前B细胞或成熟B细胞中存在Sox4缺失 细胞没有可检测到的效应。尽管有条件的淘汰赛模型使我们能够揭示 Sox4的发育阶段特异性功能，B细胞的缺乏为未来的深入研究设置了障碍 学习。为了克服这一问题，我们开发了一种体外培养系统，在该系统中，特定阶段的 发育事件取决于Sox4的存在。这一功能系统非常适合描述 Sox4功能的分子机制。我们已经进行了基因表达微阵列分析 使用培养的细胞，并确定潜在的Sox4调节基因。对Sox4进行深入的描述 在B细胞发育中的作用使用我们的新的体外系统，我们为这个项目提出了两个具体的目标。 目的1是鉴定和鉴定受Sox4直接调控的基因。目标2是描述两个角色的特征 Sox4在B细胞发育早期的下游靶点。成功完成这项拟议的研究将 阐明Sox4调控的转录程序，促进我们对B细胞发育的理解 扩展我们对PID的知识。