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; 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（由申请人提供）：该建议旨在研究由KSHV（VGPCR）ORF74编码的病毒G蛋白偶联受体的KS发病机理和KSHV生物学的作用。 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传导级联反应并诱导细胞反应，从而在体外和体内概括了KS-光型。这些包括激活VEGF介导的血管生成性和肿瘤发生。除了KSHV领域的最新进展外，本应用程序产生的工具和知识为进行VGPCR病理生物学的确定性研究提供了必要的武术，并评估其作为治疗靶点的有效性。我们的工作假设是，在裂解复制过程中的VGPCR表达具有调节和宿主病毒基因表达的能力，从而导致内皮细胞存活增加和旁分泌血管生成刺激。这些针对增强病毒复制和感染的活动在EC中具有肿瘤前和促血管生成性，因此EC中的VGPCR表达可以促进KS细胞和血管生长，也会导致致癌转化。为了研究VGPCR在KSHV感染和发病机理中的功能作用，我们将通过用趋化因子GRO-A和VGPCR-NULL突变激活KSHV感染期间VGPCR表达。为了研究KSHV在感染和复制中的作用，我们将在细胞和分子水平上进行一项确定性，VGPCR调制如何影响KSHV在293和内皮细胞中感染和复制的能力。 2-研究VGPCR表达如何影响KSHV感染期间的细胞存活，存活信号传导和KSHV基因转录。为了确定VGPCR在KSHV介导的Kaposi肉瘤发病机理中的作用，我们将研究1-使用“体外”和“体内”模型，VGPCR对KSHV感染对旁分泌激活血管生成的贡献。 2- VGPCR对KSHV感染的内皮细胞中KS样表型的概述的贡献。 3- VGPCR参与原代内皮细胞永生，转化和血管生成激活。除了测试VGPCR作为AIDS-KS疗法的靶点的有效性外，本研究还将提供在KSHV感染的背景下鉴定VGPCR致病反应的模型，并确定针对VGPCR功能的方法，能够阻止KSHV发病机理。