KSHV genome modification in KS tissue

KS 组织中的 KSHV 基因组修饰

• 批准号: 8926364
• 负责人: ERLE S. ROBERTSON
• 金额: $ 20万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-09 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8926364
• 关键词: Acetylation; Acquired Immunodeficiency Syndrome; Antibodies; Antigens; B-Lymphocytes; Cell Count; Cells; China; DNA Sequence Alteration; Developing Countries; Disease; Disease Progression; Endothelial Cells; Epigenetic Process; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Genomic Segment; Genomics; Goals; HIV; HIV Seropositivity; Histones; Human; Human Herpesvirus 8; Hypoxia; Iatrogenic Kaposi' s Sarcoma; Immunocompromised Host; Immunofluorescence Immunologic; Immunohistochemistry; Incidence; Infection; Institutes; Joints; Kaposi Sarcoma; Laboratories; Link; Lymphoma; Lytic; Malignant Neoplasms; Messenger RNA; Methylation; Modification; Molecular Profiling; Multicentric Angiofollicular Lymphoid Hyperplasia; Oncogenic; Pathway interactions; Patients; Pattern; Pleural effusion disorder; Population; Province; RNA Interference; Research; Research Personnel; Role; Signal Transduction; Spindle Endothelial Cell; Therapeutic; Time; Tissues; Transcript; Validation; Viral; Viral Genes; Viral Genome; Work; cellular targeting; chromatin immunoprecipitation; epigenetic regulation; gammaherpesvirus; insight; latent infection; lytic replication; mortality; novel; overexpression; pathogen; patient population; programs; public health relevance; therapeutic development; transcriptome sequencing; tumorigenesis

DESCRIPTION (provided by applicant): KSHV is one of the two known human oncogenic gammaherpesvirus which induces oncogenesis in infected susceptible cells. In the HIV positive immunocompromised patients, pleural effusion lymphoma (PELs), Kaposi's sarcoma (KS) and Multicentric Castleman's Disease (MCD) are common. The associated human malignancy is a major contributor to increased mortality in developing countries in this population of HIV positive patients. This study will explore epigenetic mechanism by which KSHV is regulated in endothelial cells and KS infected tissue. We will be collaborating with investigators in Shanghai and Northwest China Xinjiang province where KS incidence is quite high. We will focus on three major aims to explore the hypothesis that gene regulation in KSHV infected KS tissue is controlled by specific epigenetic changes that occur on the KSHV genome in the infected human endothelial spindle cells. We expect that genomic modifications are endothelial cell specific, and would provide clues as to the distinct profiles of expression in the KS tissue and infected endothelial cells. We will determine the specific markers which represent the epigenetic landscape in KS tissue by chromatin immunoprecipitation using specific antibodies to acetylation and methylation marks on histones. We will also identify the overall gene expression profiles related to LANA, Rta and CSL/RBP-Jk expression and the regulatory mechanisms at the viral and cellular level. Conditional expression as well as RNAi strategies for modulation of expression of LANA, Rta and CSL/RBP-Jk will be used as well as genetic mutations in the viral genomes to determine the control mechanisms that are utilized to drive proliferation of the infected KS cells. The expression profiles will be determined by RNA-Seq and analyzed for unique genes that are regulated in the viral infected cells. These will then be validated using real-time PCR analysis of the specific mRNA transcripts, as well as immunohistochemistry and immunofluorescence analysis in KS tissue and infected endothelial cells. These studies will further our understanding of KSHV latency mechanisms and provide insights for therapeutic potential by targeting the pathways activated in the KS disease.

描述（由申请人提供）：KSHV是两种已知的人类致癌γ鞘病毒之一，可在感染的易感细胞中诱导肿瘤发生。在HIV阳性免疫功能低下的患者中，胸腔积液淋巴瘤（PELS），Kaposi的肉瘤（KS）和多中心骑士氏病（MCD）很常见。相关的人类恶性肿瘤是艾滋病毒阳性人群的发展中国家死亡率增加的主要因素 患者。这项研究将探索在内皮细胞和KS感染组织中调节KSHV的表观遗传机制。我们将与上海和中国西北部新疆省的调查员合作，KS发病率很高。我们将重点介绍三个主要旨在探讨KSHV感染KS组织的基因调节的假设受到受感染人内皮内皮纺锤体细胞中KSHV基因组的特定表观遗传变化的控制。我们预计基因组修饰是内皮细胞特异性的，并且会提供有关KS组织和感染内皮细胞表达的不同表达谱的线索。我们将通过染色质免疫沉淀使用特定的乙酰化和组蛋白上的甲基化标记来确定代表KS组织中表观遗传景观的特定标记。我们还将确定与LANA，RTA和CSL/RBP-JK表达相关的总体基因表达谱以及病毒和细胞水平的调节机制。有条件的表达以及RNAi策略，用于调节LANA，RTA和CSL/RBP-JK的表达以及病毒基因组中的遗传突变，以确定用于驱动感染KS细胞增殖的控制机制。表达谱将通过RNA-seq确定，并分析在病毒感染细胞中受调节的独特基因。然后，将使用特定mRNA转录物的实时PCR分析以及KS组织和感染内皮细胞中的免疫组织化学和免疫荧光分析来验证这些。这些研究将进一步了解KSHV潜伏机制，并通过靶向KS疾病中激活的途径来提供有关治疗潜力的见解。