Project 4: Tumorigenic Effects of Tax

项目摘要 HTLV-1是成人T细胞白血病淋巴瘤（ATLL）的病原体。ATLL细胞的特征在于： 组成型NF B活化，这是其他淋巴瘤、骨髓瘤和实体瘤的关键特征。税务 癌蛋白是NF B激活的关键病毒决定因子。我们以前的研究表明， 特别是，替代性NF B途径在赋予细胞凋亡抗性中至关重要。我们的首要 假设NF B Tax活化对肿瘤发生至关重要，特别是替代（alt）NF B 通路目前的研究将使用创新的，生理淋巴瘤模型，以确定的作用， 肿瘤发生的每一个NF B途径，并确定NF B途径的关键调节。 目标1。目标1。探讨alt NF B活性在Tax介导的转化中的作用 一种新的人源化小鼠模型用于HTLV-1感染和淋巴瘤发展。我们将使用病毒 表达在激活替代NF B途径或NF B两者中具有缺陷的Tax突变体的变体 途径，以确定其在疾病发病机制中的作用。一种新的高通量病毒整合检测方法 用于监测这些实验中感染细胞的克隆性。 目标二。鉴定和表征介导alt NF B激活的Tax相互作用蛋白 我们将鉴定介导alt NF B通路激活的Tax相互作用蛋白。蛋白质与 然后将使用shRNA表征野生型而非突变型Tax，以评估它们对Tax诱导的细胞凋亡的影响。 alt NF B前体蛋白p100的切割以及对HTLV-1增殖和凋亡的影响 转化的细胞和免疫缺陷小鼠。 预期这些生理学相关的小鼠模型的信息将确定关键的靶基因 在ATLL或其他淋巴瘤的治疗试验中可能被抑制。

PROJECT SUMMARY HTLV-1 is the etiological agent of adult T-cell leukemia lymphoma (ATLL). ATLL cells are characterized by constitutive NF¿B activation, a key feature of other lymphomas, myeloma, and solid tumors. The Tax oncoprotein is the key viral determinant for NF¿B activation. Our previous studies showed that the classical and especially, the alternative NF¿B pathways were critical in conferring resistance to apoptosis. Our primary hypothesis is that Tax activation of NF¿B is critical for tumorigenesis, particularly the alternative (alt) NF¿B pathway. The current study will use innovative, physiological lymphoma models to define the role in tumorigenesis of each NF¿B pathway and identify the key regulators of the NF¿B pathway. Aim 1. Aim 1. To assess the role of alt NF¿B activity in Tax-mediated transformation A new humanized mouse model is used for HTLV-1 infection and lymphoma development. We will use viral variants expressing Tax mutants with defects in activating the alternative NF¿B pathway or both NF¿B pathways, in order to define their role in disease pathogenesis. A novel high-throughput viral integration assay is used to monitor clonality of infected cells in these experiments. Aim 2. To identify and characterize Tax interactive proteins that mediate alt NF¿B activation We will identify Tax interactive proteins that mediate alt NF¿B pathway activation. Proteins that interact with wild type but not mutant Tax will then be characterized using shRNAs to assess their effect on Tax induced cleavage of alt NF¿B precursor protein, p100, as well as effects on proliferation and apoptosis of HTLV-1 transformed cells in culture and immunodeficient mice. It is expected that the information these physiologically relevant mouse models will identify key target genes that may be inhibited in therapeutic trials of ATLL or other lymphomas.