Project 4: Tumorigenic Effects of Tax

项目总结 HTLV-1是成人T细胞白血病淋巴瘤(ATLL)的病原体。ATLL单元格的特征是 结构性核因子B激活，这是其他淋巴瘤、骨髓瘤和实体瘤的主要特征。税金 癌蛋白是核因子B激活的关键病毒决定因素。我们之前的研究表明，经典的 尤其是，可供选择的核因子B途径在抵抗细胞凋亡方面起着关键作用。我们的初选 假设核因子B的税收激活对肿瘤的发生至关重要，特别是替代(ALT)核因子B 路径。目前的研究将使用创新的、生理学的淋巴瘤模型来定义在 对每条核因子B途径的肿瘤发生进行研究，并确定核因子B途径的关键调控因子。 目的1.目的1.评估ALT核因子B活性在税收介导的转化中的作用 一种新的人源化小鼠模型被用于HTLV-1感染和淋巴瘤的发展。我们将使用病毒 表达TAX突变体的突变体，这些突变体在激活替代的NF？B途径或同时激活两者方面存在缺陷 以明确它们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 在这些实验中被用来监测感染细胞的克隆性。 目的2.鉴定和鉴定介导ALT-NF？B活化的TAX相互作用蛋白 我们将确定介导ALT NF？B途径激活的Tax相互作用蛋白。与之相互作用的蛋白质 然后将使用shRNAs表征野生型而不是突变型Tax，以评估它们对税收诱导的影响 谷丙转氨酶核因子B前体蛋白P100的切割及其对HTLV-1细胞增殖和凋亡的影响 在培养和免疫缺陷小鼠中转化细胞。 预计这些生理上相关的小鼠模型将识别关键的靶基因 这可能会在ATLL或其他淋巴瘤的治疗试验中被抑制。

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.