Histone variant H3.3 and KSHV LANA in the pathogenesis of oral Kaposi's sarcoma

组蛋白变异 H3.3 和 KSHV LANA 在口腔卡波西肉瘤发病机制中的作用

• 批准号: 8790233
• 负责人: ROLF F RENNE
• 金额: $ 22.36万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-10 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8790233
• 关键词: Accounting; Acquired Immunodeficiency Syndrome; Affect; Antibodies; Architecture; Biological Assay; Biopsy; Cell Line; Cells; ChIP-seq; Chromatin; Chromatin Structure; Clinical; Complex; DNA Tumor Viruses; DNA biosynthesis; Data; Dentistry; Deposition; Endothelial Cells; Epigenetic Process; Episome; Equilibrium; Frozen Sections; Gene Expression; Gene Expression Profile; Genetic Transcription; Genome; Goals; HIV; Herpesviridae; Herpesvirus 1; Highly Active Antiretroviral Therapy; Histone H3.3; Histones; Human; Human Herpesvirus 8; Immunofluorescence Immunologic; In Vitro; Incidence; Infection; Kaposi Sarcoma; Knock-out; Knowledge; Lymphoproliferative Disorders; Maintenance; Malignant Neoplasms; Mediating; Methods; Modification; Molecular; Molecular Chaperones; Multicentric Angiofollicular Lymphoid Hyperplasia; Mutate; Mutation; Names; Nucleosomes; Oral; Oral cavity; Palate Kaposi' s Sarcoma; Pathogenesis; Pathologist; Pathway interactions; Patients; Pilot Projects; Play; Process; Proteins; Recombinants; Recurrence; Regulation; Resolution; Role; Saliva; Site; Specificity; Specimen; System; Telomerase; Therapeutic; Time; Tissues; Transcriptional Regulation; Variant; Veins; Viral; Viral Genes; Virus; antigen binding; base; carcinogenesis; college; epigenome; experience; in vivo; innovation; latency-associated nuclear antigen; latent gene expression; lytic gene expression; lytic replication; malignant mouth neoplasm; neoplastic cell; novel; primary effusion lymphoma; public health relevance; small hairpin RNA; transmission process; tumorigenesis; vector

DESCRIPTION (provided by applicant): Kaposi's sarcoma (KS) is the most common oral cancer in human immunodeficiency virus (HIV)-infected patients. Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent of KS, and two lymphoproliferative diseases: primary effusion lymphoma (PEL) and multicentric Castleman's disease (MCD). AIDS-associated KS is highly frequent in the oral cavity which is also the major site for KSHV shedding via saliva. KSHV infected tumor cells are predominately latently infected. The viral encoded latency-associated nuclear antigen (LANA) is a multifunctional protein required for latency. Our preliminary data demonstrate that histone variant H3.3, which is frequently mutated in human malignancies and plays important roles in transcriptional regulation, occupies viral episomes at specific regions that correlate with LANA binding. We also demonstrate that LANA associates with Daxx and SSRP1, chaperone proteins responsible for the deposition of H3.3. We hypothesize that H3.3 deposition onto viral episomes is crucial for viral gene expression during latency, and furthermore that this process is mediated by LANA's interactions with H3.3 chaperones. Locus specific H3.3 incorporation may account for the tightly controlled gene expression pattern during latency and hence also affect the balance between latent and lytic replication, which may be crucial with respect to tumorigenesis and shedding in the oral cavity. Importantly, this study will not only utilize in vitro tissue cultre systems but for the first time propose a highly innovative approach to analyze complex chromatin architecture analysis in vivo using primary oral KS biopsies. To this end we provide preliminary data on our ability to detect endogenous histone H3.3 by both ChIP and IFA. The long-term goal of this interdisciplinary pilot project is to create proof of concept data that modulating histone variant deposition can be harnessed as a novel KSHV-specific therapeutic strategy.

描述(由申请人提供)： 卡波西肉瘤(KS)是人类免疫缺陷病毒(HIV)感染患者中最常见的口腔癌。Kaposi肉瘤相关疱疹病毒(KSHV)是KS和两种淋巴增生性疾病的病原体：原发渗出性淋巴瘤(PEL)和多中心Castleman病(MCD)。艾滋病相关的KS在口腔中非常常见，也是KSHV通过唾液流出的主要部位。KSHV感染的肿瘤细胞主要是潜伏感染。病毒编码的潜伏期相关核抗原(LANA)是一种潜伏期所需的多功能蛋白质。我们的初步数据表明，组蛋白变异体H3.3在人类恶性肿瘤中频繁突变，在转录调控中发挥重要作用，它占据了与LANA结合相关的特定区域的病毒表型。我们还证明了LANA与Daxx和SSRP1相关，这两种伴侣蛋白负责H3.3的沉积。我们推测，H3.3在病毒上的沉积对潜伏期病毒基因的表达是至关重要的，而且这一过程是由LANA与H3.3伴侣相互作用所介导的。位点特异性的H3.3掺入可能解释了潜伏期受到严格控制的基因表达模式，从而也影响了潜伏复制和裂解复制之间的平衡，这可能是口腔内肿瘤发生和脱落的关键。重要的是，这项研究不仅将利用体外组织培养系统，而且首次提出了一种高度创新的方法，利用初级口腔KS活检组织来分析体内复杂的染色质结构。为此，我们提供了用CHIP和IFA检测内源性组蛋白H3.3的能力的初步数据。这一跨学科试点项目的长期目标是创建概念证据数据，即调节组蛋白变异沉积可以被用作一种新的KSHV特异性治疗策略。