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B活化，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B激活的关键病毒决定因素。我们以前的研究表明，经典的，特别是替代的核因子？B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、生理学的淋巴瘤模型来确定每条核因子？B途径在肿瘤发生中的作用，并确定负责这些作用的关键核因子？B靶基因。目的1.在人源化小鼠中，哪个核因子B途径在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机制中的作用。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？B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核因子B靶基因在肿瘤进展中的作用。目的3.哪些核因子B靶基因对HTLV的转化起关键作用？在本研究中，针对核因子？B1(P105)和核因子？B2(P100)的shRNAs被用来评估每条单独的通路。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，他们选择的核因子？B靶基因对抵抗细胞凋亡的贡献将被评估。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 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. The current study will use innovative, physiological lymphoma models to define the role in tumorigenesis of each NF?B pathway and identify the key NF?B target gene responsible for these effects. Aim 1. Which NF?B 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 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 over the course o these experiments. Aim 2. Which NF?B 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 NF?B target genes in tumor progression using microarray analysis. Aim 3. Which NF?B target genes are critical for HTLV transformation? In this study, shRNAs to NF?B1 (p105) and NF?B2 (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 NF?B 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.