Role of MHV68 miRNAs in latencyand pathogenesis

MHV68 miRNA 在潜伏期和发病机制中的作用

基本信息

批准号：
8846933
负责人：
Scott A. Tibbetts
金额：
$ 36.42万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-01-01 至 2019-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8846933
关键词：
Animal Model Animals Apoptosis Attenuated B cell differentiation B-Cell Neoplasm B-Lymphocytes Binding Bioinformatics Biological Biology Cell Cycle Cell Maturation Cell Survival Conserved Sequence Coupled Development Disease Gene Targeting Genes Goals Herpesviridae Herpesviridae Infections Human Human Herpesvirus 4 Human Herpesvirus 8 Hyperplasia Immune Immune response In Vitro Individual Infection Lesion Life Link Lymphoma Lymphoproliferative Disorders Malignant - descriptor Maps Memory B-Lymphocyte Messenger RNA MicroRNAs Molecular Mus Pathogenesis Pathway interactions Phenotype Play Pneumonia Reporting Repression Research Rodent Role Seeds System Technology Testing Transcript Translations Untranslated RNA Validation Viral Virus Virus Diseases Work Xenograft Model attenuation base cancer type cellular targeting combinatorial crosslinking and immunoprecipitation sequencing deep sequencing defined contribution design drug development gammaherpesvirus in vivo in vivo Model infected B cell insight interest latent infection mouse model mutant neoplastic cell novel strategies pathogen pre-miRNA prevent programs public health relevance stem tumor

项目摘要

DESCRIPTION (provided by applicant): The transforming human gammaherpesviruses EBV and KSHV establish stable latent infections in B cells, providing a lifelong reservoir of virus tha can contribute to the development of malignant disease. Thus, defining the mechanisms that govern long-term latency in B cells is critical for designing rational strategies to prevent diseas. In vivo studies of gammaherpesviruses in humans have been severely limited by the difficulties of working in the natural host. Murine gammaherpesvirus 68 (MHV68) is related to EBV and KSHV and causes lymphomas and lymphoproliferative disease in mice, providing a readily manipulable small animal model for mechanistic studies of the virus/host relationship in vivo. Like EBV and KSHV, MHV68 expresses a defined set of miRNAs whose functions during infection are largely unknown. As these virus-encoded miRNAs are abundantly expressed in vivo in B cells during long-term latency and in hyperplastic B cell lesions during lymphoproliferative disease, we hypothesize that these viral miRNAs play key roles in latency and pathogenesis. To test this hypothesis, we will 1) Determine the functional role of individual MHV68 miRNAs in long-term latency and lymphoproliferative disease; 2) Identify specific virus and host targets of the key miRNAs using cutting-edge CLIP and deep sequencing approaches coupled with in vitro and in vivo validation; 3) Define the in vivo biological relevance of target genes and determine their roles in B cell maturation and survival. We anticipate that there will be significant overlap between the mRNA targets and cellular pathways regulated by MHV68, KSHV, and EBV miRNAs. The systematic in vivo analyses of MHV68 miRNA mutants, in conjunction with HITS-CLIP technology and in vivo validation of miRNA targets, provides an extremely powerful means to determine the molecular mechanism by which miRNAs contribute to gammaherpesvirus infection in vivo, and should allow us to define the contribution of repression of these shared targets to gammaherpesvirus latency and pathogenesis.

描述（由适用提供）：转化的人γ掌病毒EBV和KSHV在B细胞中建立稳定的潜在感染，提供终生的病毒储存库可以有助于恶性疾病的发展。这是定义控制B细胞长期潜伏期的机制对于设计预防疾病的理性策略至关重要。人类γ鞘病毒的体内研究受到自然宿主工作的困难的严重限制。鼠伽马疱疹病毒68（MHV68）与EBV和KSHV有关，并在小鼠中引起淋巴瘤和淋巴增生性疾病，为体内病毒/宿主关系的机械研究提供了易于操纵的小动物模型。像EBV和KSHV一样，MHV68表达了一组定义的miRNA，其在感染过程中的功能在很大程度上未知。由于这些病毒编码的miRNA在长期潜伏期和增生性B细胞病变中在淋巴细胞增生性疾病期间在B细胞中的体内表达绝对表达，因此我们假设这些病毒miRNA在潜伏期和发病机理中起关键作用。为了检验该假设，我们将1）确定单个MHV68 miRNA在长期潜伏期和淋巴细胞增生疾病中的功能作用； 2）使用尖端的夹子和深层测序方法与体外和体内验证结合使用，确定关键miRNA的特定病毒和宿主靶标； 3）定义靶基因的体内生物学相关性，并确定其在B细胞成熟和存活中的作用。我们预计，由MHV68，KSHV和EBV miRNA调节的mRNA靶标和细胞途径之间将存在显着重叠。 MHV68 miRNA突变体的系统体内分析，结合了MIRNA靶标的hits-clip技术和体内验证，提供了一种非常有力的方法来确定miRNA通过该分子机制，通过该机制，通过该机制，通过该机制，通过该机制有助于gammaherpesvirus在Vivo中有助于Vivo，并应允许我们定义这些共享目标的贡献和gama的贡献。