Functional analysis of KSHV GPCR in human lymphocytes

人淋巴细胞中 KSHV GPCR 的功能分析

• 批准号: 6618109
• 负责人: MARK L CANNON
• 金额: $ 11.45万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6618109
• 关键词: G protein; Kaposi's sarcoma; angiogenesis; apoptosis; biological signal transduction; cell cycle; cell line; cell migration; cell proliferation; clinical research; cytokine; cytokine receptors; flow cytometry; human genetic material tag; human herpesvirus 8; human tissue; hyperplasia; lymph nodes; lymphoma; neoplastic cell; phenotype; postdoctoral investigator; protein structure function; receptor coupling; receptor expression; virus protein

DESCRIPTION (provided by applicant): Kaposi?s sarcoma-associated herpesvirus (KSHV/HHV-8) is closely associated with Kaposi?s sarcoma (KS), primary effusion lymphoma (PEL), and multicentric Castleman?s disease, disorders associated with HIV infection. KSHV encodes a G protein-coupled receptor (GPCR) most homologous to the human IL-8 receptors CXCR1 and CXCR2. Transcription of vGPCR has been shown in KS and PEL, and evidence in transfected animal cell lines suggests that KSHV GPCR may be involved in KSHVmediated angiogenesis and oncogenesis. vGPCR has not, however, been studied in the context of PEL cell lines or any other human cell of hematopoietic origin. Since signaling molecules behave differently in different cellular contexts, such work will be fundamental in understanding the role of vGPCR in KSHV-mediated disease. To this end, Dr. Cannon designed a novel single plasmid construct that permitted development of PEL cell lines that can be made to over-express vGPCR in a dose-dependent manner using tetracycline. Preliminary data in these cell lines already show that vGPCR has quite different downstream effects than in other cell lines. Namely, vGPCR causes activation of the mitogen-activated ERK2, downregulation of stress-related p38, and decreased cell viability. Furthermore, vGPCR upregulates NFkB and AP-1 activity, two transcription factors intimately involved in B cell physiology. Importantly, other KSHV genes have been shown to have AP-1 and NFkB-responsive promoters. These cell lines are ideal tools to study the following: 1) vGPCR signaling mechanisms and downstream effects on PEL cell proliferation, cell cycle and apoptosis, 2) vGPCR-mediated autocrine/paracrine effects including secretion of VEGF, bFGF, IL-6 and other cytokines known to play a role in angiogenesis and PEL/KS biology, 3) vGPCR-mediated effects on KSHV gene transcription and life cycle. Another major aim of the proposal is to provide didactic and research components in a phased manner to provide the applicant with effective training as a physician-scientist. Dr. Cannon will be studying at the Weill Medical College and School of Medical Sciences of Cornell University and working in the lab under the mentorship of Dr. Ethel Cesarman of the Department of Pathology.

描述(申请人提供)：卡波西？S肉瘤相关疱疹病毒 (KSHV/HHV-8)与卡波西？S肉瘤(KS)密切相关 渗出性淋巴瘤与多中心性Castleman？S病 与艾滋病毒感染有关。KSHV编码G蛋白偶联受体 (GPCR)与人IL-8受体CXCR1和CXCR2最同源。 在KS和PEL中已经显示了vGPCR的转录，并且有证据表明 转基因动物细胞株提示KSHV GPCR可能参与了KSHV的介导 血管生成和肿瘤发生。然而，vGPCR还没有被研究过 在PEL细胞系或任何其他人类造血细胞的背景下 起源。由于信号分子在不同细胞中表现不同 在上下文中，这些工作将是理解vGPCR在 KSHV介导性疾病。为此，坎农博士设计了一首新颖的单曲 构建允许PEL细胞系发展的质粒组 使用四环素以剂量依赖的方式过度表达vGPCR。 在这些细胞系中的初步数据已经表明vGPCR具有相当的 与其他细胞系不同的下游效应。也就是说，vGPCR导致 丝裂原激活的ERK2，下调应激相关 P38，细胞存活率下降。此外，vGPCR上调NFkB和 AP-1活性，两种与B细胞密切相关的转录因子 生理学。重要的是，其他KSHV基因已被证明具有AP-1和 NFkB反应启动子。这些细胞系是研究细胞的理想工具。 1)vGPCR信号转导机制及其下游对PEL细胞的影响 增殖、细胞周期和细胞凋亡，2)vGPCR介导的自分泌/旁分泌 影响包括分泌血管内皮生长因子、碱性成纤维细胞生长因子、白介素6和其他已知的细胞因子 在血管生成和PEL/KS生物学中发挥作用，3)vGPCR介导的血管生成效应 KSHV基因转录与生命周期。该提案的另一个主要目的是 以分阶段的方式提供教学和研究组件，以提供 经过有效培训成为内科科学家的申请者。坎农博士会 就读于威尔医学院和中国医学院 康奈尔大学，在埃塞尔博士的指导下在实验室工作 病理学系的切萨曼。