Analysis of Cellular Events that Allow Papillomavirus Replication

允许乳头瘤病毒复制的细胞事件分析

• 批准号: 7540511
• 负责人: Elizabeth A White
• 金额: $ 4.68万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7540511
• 关键词: Antibodies; Biological Models; Cells; Chromatin; Condition; DNA Tumor Viruses; Development; Disease; Episome; Epithelial Cells; Event; Gene Expression; Genes; Genome; Goals; Histones; Human; Human Biology; Human Papillomavirus; Human papilloma virus 31; Human papilloma virus infection; Infection; Lead; Learning; Life Cycle Stages; Link; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Mediating; Microarray Analysis; Modification; Molecular; Papillomavirus; Pattern; Promoter Regions; Proteins; Public Health; Risk; Staging; Transcript; Undifferentiated; Viral; Viral Genome; Viral Physiology; Virus; Virus Replication; Woman; cell type; chromatin immunoprecipitation; human disease; interest; keratinocyte; keratinocyte differentiation; programs; promoter; research study

DESCRIPTION (provided by applicant): Human papillomaviruses (HPV) are small DNA tumor viruses that are a significant cause of human disease, and nearly every cause of cervical cancer is caused by HPV. Initial infection with HPV is extremely cell-type specific and the normal replication cycle of HPV is tightly linked to the differentiation program of the host cell, but the cellular factors responsible for these links are not yet determined. I hypothesize that specific changes in the pattern of gene expression during cellular differentiation allow papillomavirus replication to progress. This project aims to identify these changes and to determine whether they are required to promote papillomavirus replication. Since HPV infection is directly linked to the development of cervical cancer, it is important to learn about the normal life cycle of this virus so that the altered viral functions that lead to disease can be understood. The first goal of the project is the definition and functional analysis of the state of chromatin modifications at human papillomavirus promoters in differentiated versus undifferentiated cells. The second aim is to identify and analyze cellular factors that are differentially expressed during cellular differentiation and allow human papillomavirus replication. The model system to be used consists of human keratinocytes that harbor the high-risk HPV31 genome as a multi-copy episome and can be induced to differentiate using specialized culture conditions. To understand how histone modifications at the two critical HPV promoters are different before and after keratinocyte differentiation, I will perform chromatin immunoprecipitations using antibodies directed against acetylated or methylated histones in undifferentiated versus differentiated cells. In complementary experiments, I will use microarray analysis to examine the changing pattern of cellular transcripts expressed in primary keratinocytes, with or without transfected viral genomes. I will study the genes and chromatin modifications of interest identified in these assayswith the goal of understanding which differentiation-mediated changes in expression are important in allowing the later stages of HPV gene expression and replication to occur. PUBLIC HEALTH RELEVANCE: This project is directly relevant to public health. The goal of the proposed experiments is to understand the basic biology of human papillomavirus, the cause of cervical cancer. This study will provide information about how human papillomavirus replicates and about how its replication could be blocked.

描述（由申请人提供）：人乳头瘤病毒（HPV）是小的DNA肿瘤病毒，是人类疾病的重要原因，几乎所有宫颈癌的原因都是由HPV引起的。 HPV的初始感染是非常特异性的细胞类型，HPV的正常复制周期与宿主细胞的分化程序紧密相关，但是尚未确定负责这些联系的细胞因子。我假设细胞分化过程中基因表达模式的特定变化使乳头瘤病毒复制能够进展。该项目旨在确定这些变化，并确定是否需要促进乳头瘤病毒复制。由于HPV感染与宫颈癌的发展直接相关，因此了解该病毒的正常生命周期非常重要，以便可以理解导致疾病的病毒功能的改变。该项目的第一个目标是对分化和未分化细胞中人乳头瘤病毒启动子的染色质修饰状态的定义和功能分析。第二个目的是识别和分析在细胞分化过程中差异表达并允许人乳头瘤病毒复制的细胞因子。使用的模型系统由人类角质形成细胞组成，该角质形成细胞将高风险的HPV31基因组作为多拷贝曲线组成，并可以使用特殊的培养条件来诱导分化。为了了解两个关键HPV启动子的组蛋白在角质细胞分化之前和之后如何不同，我将使用针对未分化和分化细胞中针对乙酰化或甲基化组蛋白的抗体进行染色质免疫沉淀。在互补实验中，我将使用微阵列分析来检查在原代角质形成细胞中表达的细胞转录物的变化模式，无论是否有或没有转染的病毒基因组。我将研究在这些测定中鉴定的基因和染色质修饰，以了解哪种分化介导的表达变化对于允许HPV基因表达和复制的后期阶段很重要。公共卫生相关性：该项目与公共卫生直接相关。拟议的实验的目的是了解人乳头瘤病毒的基本生物学，这是宫颈癌的原因。这项研究将提供有关人类乳头瘤病毒如何复制以及如何阻止其复制的信息。