Lymphoid Transformation with Human Herpesvirus 8 K1

描述(由申请人提供)：在这份K22奖项申请中，PI通过一个全面的研究计划来描述职业发展计划，以确定HHV-8 K1在淋巴细胞和淋巴瘤中的作用。PI产生的数据表明，K1基因编码一种具有基于免疫受体酪氨酸的激活基序(ITAM)的跨膜蛋白。K1能刺激核因子-kappaB的活性，K1在转基因小鼠中的表达可诱导淋巴瘤的发生。表达K1的淋巴瘤细胞对Fas抗体诱导的细胞凋亡具有抵抗力。配备了所描述的试剂和模型，PI将能够完成拟议的研究，展示K1在淋巴细胞信号传递和可能的转化中的作用。需要检验的假设是，K1在淋巴细胞和淋巴瘤细胞中的表达刺激了NF-kappaB信号和其他途径，导致淋巴细胞转化和淋巴瘤的发展。 具体目的1.确定K1的表达是否与淋巴瘤的发生发展有关。将开发新的转基因小鼠品系，并对由此产生的淋巴瘤进行表征。 具体目的2.研究K1在淋巴瘤细胞和淋巴细胞中的信号转导途径。使用针对NF-kappaB、NFAT或AP-1的显性负性构建物和特定的活性阻断试剂，将确定K1信号的模式。ITAM缺失的K1等构建物将用于鉴定K1‘S信号通路。 具体目的3.确定K1是否抑制细胞凋亡。早期的Fas依赖的细胞凋亡的介体将通过监测caspase的激活来表征。 K1可以诱导淋巴细胞信号，这可能是导致淋巴细胞转化的早期步骤。通过分析K1信号在淋巴细胞中的表达及其在转基因小鼠中的长期表达，我们将确定K1‘S在介导细胞信号转导和淋巴瘤发生中的作用。通过执行这一应用程序中的计划，PI将展示病毒基因如何参与诱导淋巴瘤并为治疗提供见解。

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.