MyD118/CR6/GADD45 in Hematopoietic Homeostasis

描述(申请人提供)：我们研究的长期目标是 更好地了解调控体内稳态的分子调控机制 造血细胞。本研究方案以MyD118/GADD45/CR6为研究对象 基因家族，在负生长控制中发挥作用。MYD 118/GADD45/CR6家族成员也可被遗传毒性药物迅速诱导 AS通过终末分化和凋亡细胞因子，以及蛋白质 由这些基因编码的基因在负生长控制中起着关键作用。 MyD118和GADD45基因缺失小鼠的造血组织检测 显示出细胞密度增加，存活率提高，以及 未成熟细胞的比例。MYD 118/GADD45/CR6基因的非调控表达 髓系细胞系似乎加速了终末分化程序 以及与之相关的细胞凋亡。我们的工作假设是MYD 118/GADD45/CR6 通过调控发挥造血组织动态平衡的调节作用 细胞周期和细胞凋亡；表达的变化 MyD118/GADD45/CR6将改变细胞周期控制和存活，但也将 通过改变不同血统和不同阶段的分布来表现自己 造血细胞的成熟。这项研究的具体目的 建议如下：目标1：MyD118/GADD45/CR6的损失分析。小鼠品系 MyD118和GADD45为空，杂合子CR6缺失小鼠 已生成。MyD118/GADD45/CR6表达受阻的后果 在体内和体外，将对造血室进行评估。 短期和长期文化，以及对TGFb的回应，以及 压力。MyD118/GADD45/CR6表达缺失对小鼠免疫功能的影响 与癌基因c-myc、c-myb、E2F-1或ras协同作用，改变生长和/或 将确定初级BM的区别。目标2：分析 MyD118/GADD45/CR6基因表达下调。造血细胞系 已建立快速诱导解除管制的MyD118/GADD45/CR6 分析MyD118/GADD45/CR6在生长和分化中的作用以及 以应对压力和TGFb。目标3：分析 MyD11S/GADD45/CR6在凋亡途径中的作用，使用无细胞系统。无细胞 系统将有助于破译细胞凋亡，并提供一条辅助途径 MyD118、GADD45、CR6基因功能分析 相互作用的蛋白质，我们可以专注于细胞凋亡，从细胞中分离出来 周期拘禁。这些调查应该有助于更好地了解 与不同白血病的发病机制有关的遗传事件 对化疗和放射治疗的反应，最终有助于诊断， 预后和治疗。

DESCRIPTION (provided by applicant): The long-term goal of our research is to gain a better understanding of the molecular controls regulating homeostasis of hematopoietic cells. This research proposal focuses on the MyD118/Gadd45/CR6 gene family, which plays a role in negative growth control. The MyD 118/Gadd45/CR6 family members are rapidly induced by genotoxic agents, as well as by terminal differentiation and apoptotic cytokines, and the proteins encoded by these genes play pivotal roles in negative growth control. Examination of hematopoietic tissues in both MyD118 null and Gadd45 null mice revealed increased cellularity, enhanced survival and an increase in the proportion of immature cells. Deregulated expression of MyD 118/Gadd45/CR6 in myeloid cell lines appeared to accelerate the terminal differentiation program and its associated apoptosis. Our working hypothesis is that MyD 118/Gadd45/CR6 play a role in regulating homeostasis of hematopoietic tissues by modulating both the cell cycle and apoptosis; alterations in expression of MyD118/Gadd45/CR6 will modify cell cycle controls and survival, but also will manifest itself by changing the distribution of different lineages and stages of maturation of hematopoietic cells. The specific aims of this research proposal are: AIM 1: Analysis of loss of MyD118/Gadd45/CR6. Mouse strains that are null for MyD118 and Gadd45, and heterozygous CR6 null mice have been generated. The consequences of blocked expression of MyD118/Gadd45/CR6 within the hematopoietic compartment will be assessed, in vivo and in vitro, in both short term and long term cultures, as well as in response to TGFb, and to stress. The effect of loss of MyD118/Gadd45/CR6 expression on the ability to cooperate with oncogenes c-myc, c-myb, E2F-1 or ras, to alter growth and/or differentiation of primary BM will be determined. AIM 2: Analysis of deregulated expression of MyD118/Gadd45/CR6. Hematopoietic cell lines that express inducible deregulated MyD118/Gadd45/CR6 have been established to dissect the role of MyD118/Gadd45/CR6 in growth and differentiation, as well as in response to stress and TGFb. AIM 3: Analysis of the role for MyD11S/Gadd45/CR6 in apoptotic pathways, using cell free systems. Cell free systems will facilitate deciphering apoptosis, and provide an adjunct avenue of investigation to analyze the functions of MyD118, Gadd45, and CR6, as well as interacting proteins, where we can focus on apoptosis, segregated from cell cycle arrest. These investigations should contribute to a greater understanding of the genetic events involved in the pathogenesis of different leukemias and the response to chemo- and radiation therapy, ultimately aiding in diagnosis, prognosis and therapy.