Roles of MicroRNAs in KSHV Persistence and Pathogenesis

MicroRNA 在 KSHV 持续性和发病机制中的作用

• 批准号: 8740972
• 负责人: Jae U Jung
• 金额: $ 36.25万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-13 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8740972
• 关键词: AIDS related cancer; Acquired Immunodeficiency Syndrome; Address; Animal Model; Apoptosis; Biological Models; Cell Survival; Cell physiology; Cells; Cyclin-Dependent Kinase Inhibitor; Development; Disease; Functional RNA; Genes; Genetic; Genome; Herpesviridae; Herpesviridae Infections; Human Herpesvirus 8; Immune response; Individual; Infection; Intervention; Kaposi Sarcoma; Knowledge; Malignant Neoplasms; Mediating; Mesenchymal Stem Cells; Methods; MicroRNAs; Modeling; Molecular; Morbidity - disease rate; Multicentric Angiofollicular Lymphoid Hyperplasia; Mus; Mutate; Mutation; Neoplasm Metastasis; Nude Mice; Oncogenic; Pathogenesis; Pathway interactions; Patients; Phase; Prevention; Research Personnel; Role; Site; Small RNA; Societies; System; Technology; Testing; Therapeutic; Time; Transcription Coactivator; Variant; Viral; Viral Cancer; Viral Genes; Viral Physiology; Virus; Virus Diseases; Virus Latency; Work; angiogenesis; base; cell growth; cellular targeting; innovation; insight; lytic replication; metaplastic cell transformation; mortality; mutant; novel; nuclease; overexpression; positional cloning; primary effusion lymphoma; receptor; tumor; tumorigenesis

Kaposi's sarcoma-associated herpesvirus (KSHV) is associated with several AIDS-related cancers including Kaposi's sarcoma and primary effusion lymphoma. Despite intensive studies, the mechanisms underlying KSHV oncogenesis and persistent infection remain unclear. KSHV encodes more than two dozens microRNAs (miRs) derived from 12 precursor miRs. Others and we have shown that KSHV miRs regulate cell growth and survival, enhance cell invasion and angiogenesis, evade host immune responses, and promote viral latency. However, most of these studies have been carried out by overexpressing miRs without taking into consideration of KSHV infection. The objective of Project 3 is to identify the specific miRs and mechanisms that mediate KSHV oncogenesis and persistent infection in the context of viral infection. Our preliminary results have shown that a cluster of 10 KSHV pre-miRs is required for KSHV cellular transformation of primary mesenchymal stem cells (MSCs). Furthermore, we have shown that KSHV miR-K1 activates the NF- KB pathway and inhibits lytic replication by targeting IKBO while miR-KIO variants inhibit TGF-P pathway to block apoptosis by targeting TGF-(5 type II receptor. Therefore, our working hypothesis is that specific KSHV mlRs manipulate essential cellular pathways and key viral genes, contributing critically to KSHV oncogenesis and persistent infection. We have developed several novel systems that can address these challenges including model of KSHV cell growth transformation and tumorigenesis, model of KSHV infection in NOD/SCID lL2Ry-/- (NSG) "humanized" mice, KSHV reverse genetics system, and transcription activator-like effector nucleases (TALEN)-mediated genome editing technology. We will carry out the following three integrated and synergistic Specific Aims: 1. To identify KSHV essential miRs for cell growth transformation and tumorigenesis; 2. To delineate the mechanisms by which KSHV miRs regulate oncogenesis; and 3. To identify KSHV essential miRs for persistent infection in NSG "humanized" mice. The proposed works are highly significant because they will, for the first time, define the functions and mechanisms of action of KSHV miRs in oncogenesis and persistent infection using innovative approaches and newly developed model systems. The study will establish a novel paradigm of oncogenesis mediated by viral subversion ofthe mlR pathway, thus providing insights into developing innovative therapeutic methods for KSHV-induced cancers and understanding the oncogenesis of other cancers.

Kaposi的肉瘤相关疱疹病毒（KSHV）与包括Kaposi的肉瘤和原发性淋巴瘤在内的几种与AIDS相关的癌症有关。尽管进行了深入的研究，但KSHV肿瘤发生和持续感染的基础机制仍不清楚。 KSHV编码衍生自12个前体miR的二十多个microRNA（miR）。其他人，我们已经表明，KSHV miR可以调节细胞生长和生存，增强细胞侵袭和血管生成，逃避宿主免疫反应并促进病毒潜伏期。但是，这些研究大多数是通过过表达miR进行的，而无需考虑KSHV感染。项目3的目的是确定在病毒感染的情况下介导KSHV肿瘤发生和持续感染的特定miR和机制。我们的初步结果表明，基本间充质干细胞的KSHV细胞转化需要10 kSHV前的簇（MSC）。此外，我们已经表明，KSHV miR-K1激活NF-KB途径，并通过靶向IKBO来抑制裂解复制，而miR-kio变体抑制TGF-P途径通过靶向TGF-（5型II型受体）阻断凋亡。假设是特定的KSHV MLR操纵必需的细胞途径和关键的病毒基因，对KSHV的肿瘤发生和持续感染产生了严重的贡献，我们已经开发了几种新型系统，这些系统可以解决这些挑战，包括KSHV细胞生长转化和肿瘤的模型，点头/scid ll2ry - / - （NSG）“人源化”小鼠，KSHV反向遗传系统和转录激活剂样效应核酸酶（TALEN）介导的基因组编辑技术，我们将执行以下三个集成和协同特定的特定目的：1确定KSHV的基本miR，用于肿瘤的生长转化和肿瘤。提出的作品非常重要，因为它们将首次使用创新方法和新开发的模型系统定义KSHV miR在肿瘤发生和持续感染中的作用和机理。这项研究将建立一个新的肿瘤发生范式，该肿瘤发生由MLR途径的病毒颠覆介导，从而为开发具有KSHV诱导的癌症的创新治疗方法的见解提供了见解，并理解了其他癌症的肿瘤。