Human Cytomegalovirus Episome Maintenance and Propagation of Genetic Damage

人类巨细胞病毒附加体的维持和遗传损伤的传播

• 批准号: 7915225
• 负责人: Teresa D. Beeson
• 金额: $ 4.72万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2011-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7915225
• 关键词: Atherosclerosis; Autoimmunity; Cell Cycle Progression; Cells; Chromosomes, Human, Pair 1; Cytomegalovirus; Cytomegalovirus Infections; Development; Disease; Endothelial Cells; Episome; Fibroblasts; Genetic; Genome; Growth; Herpesviridae; Human; Immunocompetent; In Vitro; Infection; Kinetics; Lead; Maintenance; Malignant Neoplasms; Methods; Mitosis; Mutation; Nature; Phase; Population; Proteins; Research; Role; Testing; Viral; Viral Genome; Virus; cell growth; cell type; daughter cell; immunosuppressed; latent infection; public health relevance; segregation; tumorigenesis

DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) infects over 70 percent of the population worldwide and like other herpesviruses, it establishes a lifelong latent infection that upon reactivation can have devastating consequences in the very young, very old, or immunosuppressed. Recent research has demonstrated possible causal roles for HCMV in atherosclerosis, autoimmunity and tumorigenesis. In vitro studies have shown that certain HCMV proteins are mutagenic and that specific breaks in chromosome 1 are induced when fibroblasts are infected during S-phase. S-phase infected fibroblasts undergo mitosis and divide, and therefore it has been hypothesized that viral episome segregation during mitosis could result in one of the daughter cells lacking the HCMV-induced block to cell cycle progression and permitting the genetic damage to propagate. Further, in contrast to its interaction with fibroblasts, HCMV replicates in some endothelial cells without blocking cell cycle progression, providing another opportunity for propagation of virus-induced damage to the cellular genome. The overall objective of this application is to test the hypothesis that HCMV-induced damage to the genome of a host cell can be propagated to progeny cells, potentially dysregulating cell growth. Aim 1: To characterize specific HCMV-induced genetic damage in human cell types in which cell cycle progression is not inhibited by HCMV, specifically, aortic endothelial cells. Aim 2: To investigate and document the propagation of HCMV-induced genetic damage by (1) developing a "tagging" method to identify cells infected with HCMV or daughter cells thereof, irrespective of viral genome or gene product maintenance or concentration and (2) isolating cells identified to have lost viral genomes for further analysis of replication ability, growth kinetics and genetic damage. Cells will be cloned to encode a mCherry sequence that is preceded by a loxP-flanked STOP sequence such that infection with cre-encoded HCMV will induce cleavage of the STOP sequence and mCherry will be expressed. PUBLIC HEALTH RELEVANCE: This application seeks to document HCMV's ability to induce and propagate genetic damage in human cells, which may lead to cancer development and other diseases.

描述（由申请人提供）：人类巨细胞病毒（HCMV）感染全球70%以上的人口，与其他疱疹病毒一样，它建立了终身潜伏感染，一旦重新激活，可对幼儿，老年人或免疫抑制者造成毁灭性后果。最近的研究已经证明了HCMV在动脉粥样硬化、自身免疫和肿瘤发生中的可能因果作用。体外研究表明，某些HCMV蛋白具有诱变性，当成纤维细胞在s期感染时，1号染色体上的特定断裂会被诱导。s期感染的成纤维细胞进行有丝分裂和分裂，因此假设有丝分裂期间的病毒片段分离可能导致其中一个子细胞缺乏hcmv诱导的细胞周期进程阻断，从而允许遗传损伤繁殖。此外，与其与成纤维细胞的相互作用相反，HCMV在一些内皮细胞中复制而不阻断细胞周期进程，为病毒诱导的细胞基因组损伤的传播提供了另一个机会。该应用程序的总体目标是验证hcmv诱导的宿主细胞基因组损伤可以传播到后代细胞的假设，从而潜在地失调细胞生长。目的1：在细胞周期进程不受HCMV抑制的人类细胞类型（特别是主动脉内皮细胞）中，表征HCMV诱导的特异性遗传损伤。目的2：通过(1)开发一种“标记”方法来鉴定感染HCMV的细胞或其子细胞，而不考虑病毒基因组或基因产物维持或浓度；(2)分离鉴定出丢失病毒基因组的细胞，以进一步分析复制能力、生长动力学和遗传损伤。克隆细胞编码mCherry序列，该序列之前有一个loxp -侧翼的STOP序列，这样，被cre编码的HCMV感染将诱导STOP序列的切割，mCherry将被表达。公共卫生相关性：本申请旨在证明HCMV在人类细胞中诱导和传播遗传损伤的能力，这可能导致癌症发展和其他疾病。