Lymphoid Transformation with Human Herpesvirus 8 K1

描述（由申请人提供）： 在K22奖申请中，PI通过全面的研究计划描述了职业发展计划，以确定HHV-8 K1在淋巴细胞和淋巴瘤中的作用。 PI生成的数据显示，K1基因编码具有基于免疫受体酪氨酸的激活基序（ITAM）的跨膜蛋白。 K1刺激NF-κ B活性，转基因小鼠中K1表达诱导淋巴瘤发展。 表达K1的淋巴瘤细胞对Fas抗体诱导的细胞凋亡具有抗性。 配备所述试剂和模型后，PI将能够完成拟议的研究，显示K1在淋巴细胞信号传导和可能的转化中的作用。 待检验的假设是淋巴细胞和淋巴瘤细胞中的K1表达刺激NF-κ B信号传导和其他途径，导致淋巴细胞转化和淋巴瘤的发展。 具体目标1。 确定K1的表达是否与淋巴瘤的发生有关。 将开发新的转基因小鼠品系，并对由此产生的淋巴瘤进行鉴定。 具体目标2。 阐明K1在淋巴瘤细胞和淋巴细胞中的信号通路。 使用显性阴性构建体和靶向NF-κ B、NFAT或AP-1的特异性活性阻断剂，将确定K1信号传导的模式。 ITAM缺失的K1和其他构建体将用于鉴定K1的信号通路。 具体目标3。 确定K1是否抑制细胞凋亡。 将通过监测半胱天冬酶激活来表征fas依赖性细胞凋亡的早期介质。 K1可以诱导淋巴细胞信号传导，这可能构成导致淋巴细胞转化的早期步骤。 通过分析淋巴细胞中K1信号及其在转基因小鼠中的长期表达，我们将确定K1在介导细胞信号和淋巴瘤发展中的作用。 通过执行本申请中的计划，PI将显示病毒基因如何参与诱导淋巴瘤并提供治疗见解。

DESCRIPTION (provided by applicant): In this K22 award application, the PI describes plans for career development through a comprehensive research plan to define the role of HHV-8 K1 in lymphocytes and lymphoma. The PI has generated data showing that the K1 gene codes for a transmembrane protein with an immunoreceptor tyrosine-based activation motif (ITAM). K1 stimulates NF-kappaB activity and K1 expression in transgenic mice induces lymphoma development. Lymphoma cells expressing K1 became resistant to apoptosis that is induced by fas antibody. Equipped with the reagents and models described, the PI will be able to complete the proposed studies showing the role of K1 in lymphocyte signaling and possible transformation. The hypothesis to be tested is that K1 expression in lymphocytes and lymphoma cells stimulates NF-kappaB signaling and other pathways, leading to the transformation of lymphocytes and development of lymphoma. Specific Aim 1. To establish whether K1 expression is associated with the development of lymphoma. New lines of transgenic mice will be developed and the resulting lymphomas characterized. Specific Aim 2. To delineate the signaling pathway of K1 in lymphoma cells and lymphocytes. Using dominant negative constructs and specific active blocking reagents that target NF-kappaB, NFAT, or AP-1, the pattern of K1 signaling will be determined. ITAM deleted K1 and other constructs will be used to identify K1's signaling pathway. Specific Aim 3. To determine whether K1 suppresses apoptosis. Early mediators of fas-dependent apoptosis will be characterized through monitoring of caspase activation. K1 can induce lymphocyte signaling that may constitute the early steps leading to lymphocyte transformation. By analyzing K1 signaling in lymphocytes and its long-term expression in transgenic mice, we will determine K1's role in mediating cell signaling and lymphoma development. By carrying out the plans in this application, the PI will show how a viral gene participates in inducing lymphoma and offer insights into therapy.