Regulation of gamma herpesvirus latency & tumorigenesis

γ疱疹病毒潜伏期的调节

• 批准号: 6745864
• 负责人: HERBERT W VIRGIN
• 金额: $ 35.96万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6745864
• 关键词: B lymphocyte; BCL2 gene /protein; cytotoxic T lymphocyte; gene mutation; host organism interaction; human herpesvirus 8; laboratory mouse; latent virus infection; viral carcinogenesis; virus genetics

DESCRIPTION (provided by applicant): This is a revised (score 159/26.8%) competing renewal for RO1 CA74730 "Regulation of gammaherpesvirus latency and tumorigenesis" which funded studies of the mechanisms of y-herpesvirus latency using infection with murine gamma-herpesvirus 68 (gammaHV68) as a model. We defined both host immune mechanisms (B cells, antibody, CD8 T cells, IFNalphabeta, STAT1, IFNgamma, perforin, caspase 3, granzymes) and viral genes (M1, M2, gene 50, gene 73 (KSHV LANA homolog), v-bcI-2, v-cyc/in, and v-GPCR) that control persistent replication and latency. Particularly interesting was the role for the v-Bcl-2, which is conserved across gamma-herpesviruses, in latency and persistent infection. We also developed, as proposed, a robust model for gammaHV68 induction of B cell lymphomas. Here we propose to continue identification of the genes critical for latency using newly developed methods for unbiased identification of candidate herpesvirus latency transcripts combined with loss of function mutagenesis. Taking advantage of the new tumor model we will address the fundamentally important question of whether latency and lymphomagenesis require the same genes. To understand the cellular basis of gammaHV68 latency, we will define the effects of gammaHV68 infection and expression of latency associated genes on the proliferation and survival of primary B cells. To understand latency at the molecular level, we will define the mechanisms by which the v-Bcl-2 regulates ?HV68 latency. To this end we have generated new data identifying a host apoptotic pathway (perforin/granzymes) that regulates gammaHV68 latency, performed structural and biochemical characterizations of the KSHV, EBV, and gammaHV68 v-Bcl-2 proteins, and shown that the conserved v-Bcl-2 BH1 domain is important for latency. These data provide a firm foundation for experiments identifying how the v-Bcl-2 protein contributes to chronic ?HV68 infection. These interrelated goals will be pursued through three Aims: Aim 1: Identify the role of latency and tumor associated genes in latency and lymphomagenesis. Aim 2: Identify the role of latency and tumor-associated genes in B cell proliferation and survival. Aim 3: Identify the mechanism by which v-Bcl-2 regulates gammaHV68 latency.

描述（由申请人提供）：这是 RO1 CA74730“γ-疱疹病毒潜伏期和肿瘤发生的调节”的修订版（得分 159/26.8%）竞争性更新，资助使用鼠γ-疱疹病毒 68 感染来研究 y-疱疹病毒潜伏期机制（gammaHV68）作为模型。我们定义了宿主免疫机制（B 细胞、抗体、CD8 T 细胞、IFNalphabeta、STAT1、IFNgamma、穿孔素、caspase 3、颗粒酶）和病毒基因（M1、M2、基因 50、基因 73（KSHV LANA 同源物）、v- bcI-2、v-cyc/in 和 v-GPCR）控制持久复制和延迟。特别有趣的是 v-Bcl-2 在潜伏和持续感染中的作用，它在 γ-疱疹病毒中是保守的。正如所提议的，我们还开发了一个用于 gammaHV68 诱导 B 细胞淋巴瘤的稳健模型。在这里，我们建议使用新开发的方法继续鉴定对潜伏期至关重要的基因，以公正地鉴定候选疱疹病毒潜伏期转录本并结合功能丧失诱变。利用新的肿瘤模型，我们将解决潜伏期和淋巴瘤发生是否需要相同基因的根本性重要问题。为了了解 gammaHV68 潜伏期的细胞基础，我们将定义 gammaHV68 感染和潜伏期相关基因的表达对原代 B 细胞增殖和存活的影响。为了在分子水平上了解潜伏期，我们将定义 v-Bcl-2 调节 ?HV68 潜伏期的机制。为此，我们生成了新数据，确定了调节 gammaHV68 潜伏期的宿主细胞凋亡途径（穿孔素/颗粒酶），对 KSHV、EBV 和 gammaHV68 v-Bcl-2 蛋白进行了结构和生化表征，并表明保守的 v- Bcl-2 BH1 域对于延迟很重要。这些数据为确定 v-Bcl-2 蛋白如何促进慢性 HV68 感染的实验提供了坚实的基础。这些相互关联的目标将通过三个目标来实现： 目标 1：确定潜伏期和肿瘤相关基因在潜伏期和淋巴瘤发生中的作用。目标 2：确定潜伏期基因和肿瘤相关基因在 B 细胞增殖和存活中的作用。目标 3：确定 v-Bcl-2 调节 gammaHV68 潜伏期的机制。