PROLIFERATIVE EFFECTS OF HTLV-1

描述(申请人提供)：HTLV-1是成人T细胞白血病淋巴瘤(ATLL)的病原体。ATLL细胞的特征是结构性的核因子κB激活，这是其他淋巴瘤、骨髓瘤和实体瘤的关键特征。Tax癌蛋白是核因子κB激活的关键病毒决定因素。我们以前的研究表明，经典的，特别是替代的NFκB途径在诱导细胞凋亡抵抗中起着关键作用。目前的研究将使用创新的生理性淋巴瘤模型来确定每一条NFκB途径在肿瘤发生中的作用，并确定负责这些影响的关键NFκB靶基因。目的1.哪个核因子κB途径在人源化小鼠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被用于HTLV-1感染和淋巴瘤的发生。我们将使用表达TAX突变体的病毒突变体来确定它们在疾病发病中的作用。这些突变体在激活替代的NFκB途径或同时激活两条NFκB途径方面存在缺陷。在这些实验过程中，一种新的高通量病毒整合试验被用来监测感染细胞的克隆性。目的2.TAX转基因肿瘤的维持和发展需要哪些核因子κB靶点？在这项研究中，利用一种新的可诱导的淋巴瘤转基因小鼠模型，利用微阵列分析来评估特定的NFκB靶基因在肿瘤进展中的作用。目的3.哪些核因子κB靶基因对烟草花叶病毒的转化起关键作用？在本研究中，针对NFκB1(P105)和NFκB2(P100)的shRNA被用来评估每一条单独的途径。表达这两种shRNA的HTLV-1转化细胞将接受微阵列分析，以确定哪条途径负责特定靶基因的激活。此外，还将评估他们选择的NFκB靶基因对抵抗细胞凋亡的贡献。预计这些生理上相关的小鼠模型将识别在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.