Cell model for KSHV infection and genetic manipulation

KSHV 感染和基因操作的细胞模型

基本信息

批准号：
8330348
负责人：
Shou-Jiang Gao
金额：
$ 37.1万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-01-13 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8330348
关键词：
3&apos Untranslated Regions AIDS related cancer Acquired Immunodeficiency Syndrome Algorithms Artificial Chromosomes Bacteria Biological Assay Biological Models Biological Process Biological Sciences Cell Communication Cell Proliferation Cell model Cells Collaborations Complex Computational algorithm DNA biosynthesis Data Analyses Development Disease Endothelial Cells Funding Gene Expression Gene Mutation Genes Genetic Genetic Models Genome Goals Health Herpesviridae Herpesviridae Infections Human Human Herpesvirus 8 Individual Infection Inflammation Kaposi Sarcoma Lead Light Lymphoma Lymphoproliferative Disorders Lytic Malignant - descriptor Malignant Neoplasms Mediating Metabolism MicroRNAs Modeling Molecular Molecular Genetics Molecular Profiling Morbidity - disease rate Multicentric Angiofollicular Lymphoid Hyperplasia Mutagenesis Nature Outcome Pathogenesis Pathway interactions Patients Phase Phenotype Prevention Production Proteins Proteomics Recombinants Regulation Reporter Research Personnel Role Seeds Series Signal Transduction Societies Staging System Systems Biology Testing Time Umbilical vein Viral Viral Genes Viral Genome Viral Physiology Virion Virus Virus Diseases Virus Latency Work angiogenesis base cellular targeting effusion frontier genetic manipulation innovation insight latent infection loss of function lytic replication mortality multidisciplinary mutant novel overexpression programs reactivation from latency viral DNA

项目摘要

DESCRIPTION (provided by applicant): Kaposi's sarcoma-associated herpesvirus (KSHV) is causally associated with Kaposi's sarcoma (KS) and several other AIDS-related malignancies. The long-term goal of this project is to apply effective KSHV genetic and infection systems to delineate the molecular mechanism of KSHV-induced pathogenesis. In previous funding period, we have developed and refined a KSHV genetic manipulation system and a KSHV infection model of primary human umbilical vein endothelial cell (HUVEC). We have used these model systems to successfully define the functions of KSHV genes and examine KSHV-cell interactions. These studies have shown that both KSHV latent and lytic replication phases are important for KSHV-induced malignant transformation. Thus, defining the mechanism controlling KSHV latency and lytic replication is a critical step in delineating the pathogenesis of KSHV-induced malignancies. Toward this goal, our recent studies have shown that KSHV-encoded microRNAs (miRs) promote viral latency by inhibiting viral lytic replication program. To further understand the functions of KSHV miRs, we have developed novel computational algorithms for miR target prediction and time-series data analysis for complex signaling networks. The objective of this renewal application is to continue to define the functions of KSHV miRs in viral infection and replication, and delineate the underlying molecular mechanisms. Our hypothesis is that KSHV miRs regulate viral lifecycle by targeting viral and/or cellular genes to inhibit specific stages of KSHV infection and replication. To test this hypothesis and accomplish the objective, we will carry out the following three specific aims: (1) To define the stages of viral infection and replication regulated by KSHV miRs by examining virus production, viral gene expression, viral DNA replication, and virion packaging and egress; (2) To identify the specific viral miRs that regulate KSHV infection and replication by genetic complementation and loss-of-function approaches using miR suppressors or genetic mutants; and (3) To delineate the mechanisms by which KSHV miRs regulate viral infection and replication by identifying the direct viral and cellular targets using a systems biology approach. The proposed project is significant because it will, for the first time, define the roles of KSHV miRs in viral infection and replication, and delineate the underlying mechanisms. The proposed study is multidisciplinary in nature, and innovative through the integration of novel computational, genetic, molecular, cellular, microarray and proteomic approaches. The successful completion of this project will lead to the establishment of a novel systems biology approach for delineating the functions of viral and cellular miRs using KSHV as a model system. The outcomes will not only provide insights into the mechanism of KSHV latency and replication but also shed light on the pathogenesis of KSHV-induced malignancies. PUBLIC HEALTH RELEVANCE: Kaposi's sarcoma is a common malignancy in AIDS patients in US and worldwide inflicting morbidity and mortality to the society. This project will investigate the mechanism underlining the development of Kaposi's sarcoma, and identify potential targets for the prevention and treatment of this disease.

描述（由申请人提供）：卡波西肉瘤相关疱疹病毒（KSHV）与卡波西肉瘤（KS）和其他几种与艾滋病相关的恶性肿瘤有因果关系。该项目的长期目标是应用有效的 KSHV 遗传和感染系统来描述 KSHV 诱导发病的分子机制。在前期资助期间，我们开发并完善了KSHV基因操作系统和原代人脐静脉内皮细胞（HUVEC）的KSHV感染模型。我们已经使用这些模型系统成功定义了 KSHV 基因的功能并检查了 KSHV 与细胞的相互作用。这些研究表明，KSHV 潜伏期和裂解复制期对于 KSHV 诱导的恶性转化都很重要。因此，确定控制 KSHV 潜伏期和裂解性复制的机制是描述 KSHV 诱导的恶性肿瘤发病机制的关键步骤。为了实现这一目标，我们最近的研究表明，KSHV 编码的 microRNA (miR) 通过抑制病毒裂解复制程序来促进病毒潜伏期。为了进一步了解 KSHV miR 的功能，我们开发了新的计算算法，用于 miR 靶点预测和复杂信号网络的时间序列数据分析。此次更新申请的目的是继续定义 KSHV miR 在病毒感染和复制中的功能，并描绘潜在的分子机制。我们的假设是，KSHV miR 通过靶向病毒和/或细胞基因来抑制 KSHV 感染和复制的特定阶段来调节病毒生命周期。为了检验这一假设并实现目标，我们将实现以下三个具体目标：（1）通过检查病毒产生、病毒基因表达、病毒DNA复制和病毒颗粒来定义KSHV miRs调控的病毒感染和复制阶段包装和出口； (2) 利用miR抑制子或基因突变体，通过基因互补和功能丧失的方法，鉴定调节KSHV感染和复制的特定病毒miR； (3) 通过使用系统生物学方法识别直接病毒和细胞靶标，描述 KSHV miR 调节病毒感染和复制的机制。该项目意义重大，因为它将首次定义 KSHV miR 在病毒感染和复制中的作用，并描述其潜在机制。拟议的研究本质上是多学科的，并且通过新颖的计算、遗传、分子、细胞、微阵列和蛋白质组学方法的整合而具有创新性。该项目的成功完成将导致建立一种新的系统生物学方法，使用 KSHV 作为模型系统来描述病毒和细胞 miR 的功能。研究结果不仅将提供对 KSHV 潜伏和复制机制的见解，还将揭示 KSHV 诱发的恶性肿瘤的发病机制。公共卫生相关性：卡波西肉瘤是美国和全世界艾滋病患者的常见恶性肿瘤，给社会带来发病率和死亡率。该项目将研究卡波西肉瘤发生的机制，并确定预防和治疗该疾病的潜在靶点。