Functional analysis of KSHV GPCR in human lymphocytes

人淋巴细胞中 KSHV GPCR 的功能分析

• 批准号: 7494752
• 负责人: MARK L CANNON
• 金额: $ 5.18万
• 依托单位: UNIVERSITY OF MINNESOTA
• 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 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7494752
• 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（由申请人提供）：Kaposi是肉瘤相关的疱疹病毒 （KSHV/HHV-8）与Kaposi的Sarcoma（KS）密切相关 积液淋巴瘤（PEL）和多中心Castleman病，疾病 与HIV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。 VGPCR的转录已在KS和PEL中显示，并在 转染的动物细胞系表明KSHV GPCR可能参与KSHVSIDED 血管生成和肿瘤发生。但是，VGPCR尚未研究 在PEL细胞系或造血的任何其他人类细胞中 起源。由于信号分子在不同的细胞中的行为不同 背景下，这种工作将是理解VGPCR的作用的基础 KSHV介导的疾病。为此，坎农博士设计了一本小说单曲 允许开发PEL细胞系的质粒结构 使用四环素以剂量依赖性方式过表达VGPCR。 这些细胞系中的初步数据已经表明VGPCR具有很大的 下游效应与其他细胞系不同。即VGPCR原因 激活有丝分裂原活化的ERK2，与应力相关的下调 p38，并降低细胞活力。此外，VGPCR上调了NFKB和 AP-1活性，两个转录因子与B细胞密切涉及 生理。重要的是，其他KSHV基因已被证明具有AP-1和 NFKB响应启动子。这些细胞系是研究的理想工具 以下：1）VGPCR信号传导机制和对PEL细胞的下游影响 增殖，细胞周期和凋亡，2）VGPCR介导的自分泌/旁分泌 影响包括VEGF，BFGF，IL-6和其他已知的细胞因子的分泌 在血管生成和PEL/KS生物学中起作用，3）VGPCR介导的对 KSHV基因转录和生命周期。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 作为医生科学家有效培训的申请人。 Cannon Will博士 在威尔医学院和医学科学学院学习 康奈尔大学并在Ethel博士的指导下在实验室工作 病理学系的Cesarman。