PROLIFERATIVE EFFECTS OF HTLV-1

描述（由申请人提供）：HTLV-1是成人t细胞白血病淋巴瘤（ATLL）的病原。ATLL细胞的特征是组成性NFκB活化，这是其他淋巴瘤、骨髓瘤和实体瘤的一个关键特征。Tax癌蛋白是NFκB活化的关键病毒决定因素。我们之前的研究表明，经典的，特别是替代的NFκB通路在赋予细胞凋亡抗性方面是至关重要的。目前的研究将使用创新的生理淋巴瘤模型来确定每个NFκB通路在肿瘤发生中的作用，并确定负责这些作用的关键NFκB靶基因。目的1。哪种NFκB通路在人源化小鼠HTLV肿瘤发生中起关键作用？在这项研究中，一种新的人源化小鼠模型用于HTLV-1感染和淋巴瘤的发展。我们将使用表达Tax突变体的病毒变体，这些突变体在激活另一种NFκB途径或两种NFκB途径方面存在缺陷，以确定它们在疾病发病机制中的作用。一种新型的高通量病毒整合试验用于监测这些实验过程中感染细胞的克隆性。目标2。哪些NFκB靶点是维持和发展Tax转基因肿瘤所必需的？本研究利用一种新的诱导型Tax转基因淋巴瘤小鼠模型，通过微阵列分析来评估特异性NFκB靶基因在肿瘤进展中的作用。目标3。哪些NFκB靶基因对HTLV转化至关重要？在本研究中，使用NFκB1 （p105）和NFκB2 （p100）的shrna来评估每个单独的途径。HTLV-1转化的细胞表达这两种shrna中的一种，将进行微阵列分析，以确定哪一种途径负责特定靶基因的激活。此外，我们还将评估它们所选择的NFκB靶基因对细胞凋亡抗性的贡献。这些生理上相关的小鼠模型的信息有望确定在ATLL或其他淋巴瘤的治疗试验中可能被抑制的关键靶基因。

DESCRIPTION (provided by applicant): HTLV-1 is the etiological agent of adult T-cell leukemia lymphoma (ATLL). ATLL cells are characterized by constitutive NFκB activation, a key feature of other lymphomas, myeloma, and solid tumors. The Tax oncoprotein is the key viral determinant for NFκB activation. Our previous studies showed that the classical and especially, the alternative NFκB pathways were critical in conferring resistance to apoptosis. The current study will use innovative, physiological lymphoma models to define the role in tumorigenesis of each NFκB pathway and identify the key NFκB target gene responsible for these effects. Aim 1. Which NFκB pathway is critical for HTLV tumorigenesis in humanized mice? In this study, 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 NFκB pathway or both NFκB 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 over the course o these experiments. Aim 2. Which NFκB targets are required for maintenance and progression of Tax transgenic tumors? In this study, a new inducible Tax transgenic mouse model of lymphoma is utilized to assess the role of specific NFκB target genes in tumor progression using microarray analysis. Aim 3. Which NFκB target genes are critical for HTLV transformation? In this study, shRNAs to NFκB1 (p105) and NFκB2 (p100) are used to assess each individual pathway. HTLV-1 transformed cells expressing one of both of these shRNAs will be subjected to microarray analysis to determine which pathway is responsible for activation of specific target genes. Moreover, their contribution of select NFκB target genes to resistance to apoptosis will be assessed. 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.