Interaction of HTLV-1 Tax & Hbz in Transformation

HTLV-1 Tax与Hbz在转化过程中的相互作用 摘要 人类T细胞白血病病毒1型(HTLV-1)是一种难治性T细胞淋巴增生性疾病的原因， 成人T细胞白血病淋巴瘤(ATLL)。HTLV-1编码两个癌基因Tax和Hbz，每个基因 在转基因小鼠中诱发T细胞淋巴瘤。Tax是一种有效的转录反式激活蛋白， 也会导致遗传不稳定。然而，TAX具有免疫原性，其表达在以下情况下下调 ATIL的发展。Hbz RNA和蛋白质具有促进T细胞活化和增殖的可分离活性。 Hbz在整个感染过程中持续表达。我们假设税收的主要作用是T细胞 淋巴瘤的发生，而Hbz的主要作用是T细胞淋巴瘤的促进剂。我们开发了三个 动物模型来验证这一假说，并阐明这些作用的潜在分子机制 癌基因及其相互作用。在我们的Tax Hbz转基因小鼠中，Tax受 多西环素诱导启动子。T细胞淋巴瘤在多西环素存在的情况下发生，并在其 不在了。在我们的Tax-Hbz转基因小鼠中，全长Hbz RNA和功能表位标记的Hbz蛋白是 在活化的T细胞中表达，并诱导T细胞淋巴瘤。双转基因税务Hbz小鼠发育 肿瘤比任何一种单一转基因模型都要快。这些动物有需要监控的成像标记 多西环素诱导与税收活性。在目标1中，我们将通过以下方式比较完整的外显子基因组学和转录组学 Tax Hbz-、Tax-Hbz和Tax Hbz小鼠的RNAseq都在多西环素上维持，以确定是否 税收在我们的小鼠模型中诱导遗传不稳定，以及是否在频繁发生突变的基因中发生突变 在ATLL中发生了突变。我们将使用RNAseq来识别负责每个转基因独特特征的基因， 如果双转基因动物与单转基因动物相比有明显的变化。在AIM 2，我们将停用多西环素，以确定肿瘤是否在税务Hbz动物中继续增殖，但 通过RNAseq，确定与不同生物学特性有关的基因 结果。我们会将我们的转录改变与ATLL中的比较。在目标3中，我们将使用shRNA或 慢病毒在转基因模型中确定的关键基因的表达，以测试来自我们的 短期培养的ATHL细胞系小鼠模型的建立。这些研究将提供新的见解，了解税收和 Hbz介导的转化，可能导致atll的新疗法和新生物标记物的鉴定。 治疗这种疾病。

Interaction of HTLV-1 Tax & Hbz in Transformation Abstract Human T-cell leukemia virus type 1 (HTLV-1) is the cause of a refractory T cell lymphoproliferative disorder, designated adult T-cell leukemia lymphoma (ATLL). HTLV-1 encodes two oncogenes, Tax and Hbz, each of which induces T cell lymphoma in transgenic mice. Tax is a potent transcriptional trans activator protein that also induces genetic instability. However, Tax is immunogenic and its expression is down-regulated after ATLL develops. Hbz RNA and protein have separable activities that promote T cell activation and proliferation. Hbz is continuously expressed throughout infection. We hypothesize that the predominant role of Tax is T cell lymphoma initiation, and the predominant role of Hbz is a T cell lymphoma promoter. We developed three animal models to test this hypothesis and elucidate the underlying molecular mechanisms for the roles of these oncogenes and their interactions. In our Tax+Hbz- transgenic mice, Tax is under the regulation of a doxycycline inducible promoter. T cell lymphomas develop in the presence of doxycycline, and regress in its absence. In our Tax-Hbz+ transgenic mice, full length Hbz RNA and functional epitope tagged Hbz protein are expressed in activated T cells and induce T cell lymphomas. Double transgenic Tax+Hbz+ mice develop tumors more rapidly than either single transgenic model. These animals have imaging markers to monitor doxycycline induction and Tax activity. In Aim 1, we will compare full exon genomics and transcriptomics by RNAseq of Tax+Hbz-, Tax-Hbz+, and Tax+Hbz+ mice all maintained on doxycycline in order to determine if Tax induces genetic instability in our mouse model and whether mutations occur in genes that are frequently mutated in ATLL. We will use RNAseq to identify genes responsible for the unique features of each transgene, and if there are distinct alterations in double transgenic animals compared to single transgenic animals. In Aim 2, we will withdraw doxycycline in order to determine if tumors continue to proliferate in Tax+Hbz+ animals, but regress in Tax+Hbz- animals, and to determine, by RNAseq, the genes responsible for the different biological results. We will compare our transcriptomic alterations to those in ATLL. In Aim 3, we will use shRNAs or lentivirus expression of key genes identified in the transgenic models, to test high priority candidates from our murine model in short-term cultured ATLL cell lines. These studies will provide new insights into how Tax and Hbz mediate transformation, which could lead to new therapies for ATLL and identification of novel biomarkers for this disease.