Interaction of HTLV-1 Tax & Hbz in Transformation

HTLV-1 Tax与Hbz在转化中的相互作用 摘要 人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病因， 称为成人T细胞白血病淋巴瘤（ATLL）。HTLV-1编码两种癌基因Tax和Hbz，每种基因都是 其在转基因小鼠中诱导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是免疫原性的，其表达在转染后下调。 ATLL开发。Hbz RNA和蛋白具有促进T细胞活化和增殖的可分离的活性。 Hbz在整个感染过程中持续表达。我们假设Tax的主要作用是T细胞 Hbz的主要作用是T细胞淋巴瘤启动子。我们开发了三个 动物模型来验证这一假设，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受到一种新的调节因子的调节。 强力霉素诱导型启动子。T细胞淋巴瘤在多西环素存在下发生，并在其存在下消退。 缺席在我们的Tax-Hbz+转基因小鼠中，全长Hbz RNA和功能性表位标记的Hbz蛋白被表达。 在活化的T细胞中表达并诱导T细胞淋巴瘤。双转基因Tax+Hbz+小鼠开发 肿瘤比单一转基因模型更快。这些动物有成像标记来监测 强力霉素诱导和Tax活性。在目标1中，我们将通过以下方式比较全外显子基因组学和转录组学： Tax+Hbz-、Tax-Hbz+和Tax+Hbz+小鼠的RNAseq均维持在多西环素上，以确定是否存在与Tax + Hbz-、Tax-Hbz+和Tax +Hbz+小鼠的RNAseq相关的基因。 在我们的小鼠模型中，税收诱导了遗传不稳定性，以及突变是否发生在经常发生的基因中。 在ATLL中发生突变。我们将使用RNAseq来鉴定负责每个转基因独特特征的基因， 以及与单转基因动物相比，双转基因动物是否存在明显的变化。在Aim中 2，我们将停用强力霉素以确定肿瘤是否在Tax+Hbz+动物中继续增殖，但 在Tax+Hbz-动物中回归，并通过RNAseq确定负责不同生物学特性的基因。 结果我们将比较我们的转录组的变化，在ATLL。在目标3中，我们将使用shRNA或 在转基因模型中鉴定的关键基因的慢病毒表达，以测试来自我们的高优先级候选物。 在短期培养的ATLL细胞系中的鼠模型。这些研究将提供新的见解如何税收和 Hbz介导转化，这可能导致ATLL的新疗法和新生物标志物的鉴定 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.