BIOLOGY OF KSHV/HHV8 G PROTEIN COUPLED RECEPTOR

KSHV/HHV8 G 蛋白偶联受体的生物学

• 批准号: 7082650
• 负责人: Enrique A Mesri
• 金额: $ 34.09万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7082650
• 关键词: G protein; Kaposi' s sarcoma; SCID mouse; angiogenesis; athymic mouse; biological signal transduction; cell surface receptors; human herpesvirus 8; molecular cloning; molecular pathology; neoplastic transformation; receptor binding; receptor expression; tissue /cell culture; transfection; virus infection mechanism; virus receptors; virus replication

DESCRIPTION (provided by applicant): This proposal is directed to investigate the role in KS pathogenesis and KSHV biology of the viral G protein coupled receptor encoded by ORF74 of KSHV (vGPCR). vGPCR appear to be a critical gene in KSHV pathobiology because it can trigger important signaling cascades and induce cellular responses that recapitulate the KS-phenotype in vitro and in vivo. These include activation of VEGF mediated angiogenicity and tumorigenesis. The tools and knowledge generated by the present application in addition to recent advances in the KSHV field provide the necessary armamentarium to undertake a definitive study in vGPCR pathobiology and assess its validity as therapeutic target. Our working hypothesis is that vGPCR expression during lytic replication has the capacity to regulate and host viral gene expression leading to increased endothelial cell survival during and paracrine angiogenic stimulation. That these activities that are directed to enhance viral replication and infection are pre-neoplastic and pro-angiogenic in EC, and thus vGPCR expression in EC can promote KS-cell and vessel growth and also lead to oncogenic transformation. To study the functional role of vGPCR in KSHV infection and pathogenesis we will modulate vGPCR expression during KSHV infection by activation with the chemokine Gro-a and by vGPCR-null mutations. To study the role of KSHV in infection and replication we will 1-Determine at the cellular and molecular level how vGPCR-modulation affects the ability of KSHV to infect and replicate in 293 and endothelial cells. 2- Study how vGPCR expression affects cell survival, survival signaling and KSHV gene transcription during KSHV infection. To determine the role of vGPCR in KSHV mediated Kaposi's sarcoma pathogenesis we will study 1- The contribution of vGPCR to paracrine activation of angiogenesis by KSHV infection using "in vitro" and "in vivo" models. 2- The contribution of vGPCR to the recapitulation of KS-like phenotypes in KSHV infected endothelial cells. 3- The participation of vGPCR in primary endothelial cell immortalization, transformation and angiogenic activation. In addition to test the validity of vGPCR as target for AIDS-KS therapy, this research will provide models that identify vGPCR pathogenic responses in the context of KSHV infection and identify approaches targeted to vGPCR function that are able to block KSHV pathogenesis.

描述（由申请方提供）：本提案旨在研究KSHV ORF 74编码的病毒G蛋白偶联受体（vGPCR）在KS发病机制和KSHV生物学中的作用。vGPCR似乎是KSHV病理生物学中的关键基因，因为它可以触发重要的信号级联并诱导在体外和体内重现KS表型的细胞应答。这些包括VEGF介导的血管生成和肿瘤发生的激活。除了KSHV领域的最新进展之外，本申请产生的工具和知识提供了进行vGPCR病理生物学的确定性研究并评估其作为治疗靶标的有效性的必要工具。我们的工作假设是，裂解复制过程中的vGPCR表达具有调节和宿主病毒基因表达的能力，导致旁分泌血管生成刺激过程中内皮细胞存活增加。这些旨在增强病毒复制和感染的活性在EC中是肿瘤前和促血管生成的，因此EC中的vGPCR表达可以促进KS细胞和血管生长，并且还导致致癌转化。为了研究vGPCR在KSHV感染和发病机制中的功能作用，我们将通过用趋化因子Gro-a激活和vGPCR无效突变来调节KSHV感染期间的vGPCR表达。为了研究KSHV在感染和复制中的作用，我们将1-在细胞和分子水平上确定vGPCR调节如何影响KSHV在293和内皮细胞中感染和复制的能力。2-研究在KSHV感染期间vGPCR表达如何影响细胞存活、存活信号传导和KSHV基因转录。为了确定vGPCR在KSHV介导的卡波西肉瘤发病机制中的作用，我们将使用“体外”和“体内”模型研究1-vGPCR对KSHV感染引起的血管生成的旁分泌激活的贡献。2-vGPCR对KSHV感染的内皮细胞中KS样表型重演的贡献。3-vGPCR参与原代内皮细胞永生化、转化和血管生成激活。除了测试vGPCR作为AIDS-KS治疗靶点的有效性外，本研究还将提供识别KSHV感染背景下vGPCR致病反应的模型，并识别能够阻断KSHV发病机制的靶向vGPCR功能的方法。