Targets and functions of the Kaposi's Sarcoma associated herpesvirus microRNAs

描述（由申请人提供）：在2010年夏天，我希望转移到一个独立的教师职位，以追求卡波西肉瘤相关疱疹病毒（KSHV）microRNAs（miRNAs）的功能分析。MicroRNA是最近发现的一类长约22个核苷酸的非编码RNA，其转录后抑制与miRNA不完全匹配的mRNA，主要在其3 'UTR。KSHV在潜伏感染期间表达12种病毒miRNA。我的近期研究目标是获得KSHV miRNA靶点的详细知识，并阐明这些miRNA对宿主细胞先天免疫反应（包括细胞凋亡和细胞周期阻滞）的可能保护作用。通过结合稳定表达生理水平的一种或多种KSHV miRNA的B细胞系的基因表达分析和基于结合能的靶标预测，将鉴定KSVH miRNA介导的调控的一组广泛的候选靶标。将研究KSHV miRNA对诱导细胞凋亡和细胞周期停滞的干扰，（a）基于在这些途径中具有已知功能的高度有希望的候选靶标，（B）通过异位表达单个或多个KSHV miRNA的B细胞系或内皮细胞的系统功能询问，（c）通过对潜伏性KSHV感染的原发性渗出性淋巴瘤（PEL）细胞的系统性功能询问，其中单个或多个miRNA的功能已被使用KSHV或海绵阻断。在指导阶段，我打算通过参加杜克提供的课程来获得生物信息学方面的技能。我将通过与我的导师Bryan Cullen和计算生物学，疱疹病毒学和细胞凋亡/细胞周期专家指导委员会的定期会议来监测我进一步发展成为一名独立研究人员。此外，我希望拟议中的研究将产生足够的有趣的数据，以申请R 01赠款早期在独立阶段，也进一步确定我的长期研究计划。 相关性：相关性：致突变人类γ疱疹病毒KSHV在潜伏期内表达12种病毒mIRNA。阐明KSHV miRNAs的功能有望大大增强我们对KSHV生物学的理解，并可能为KSHV和miR-155（KSHV microRNA miR-K12-11的直系同源物）的肿瘤发生机制提供重要线索。

DESCRIPTION (provided by applicant): In the summer of 2010,1 am hoping to move to an independent faculty position to pursue the functional analysis of Kaposi's sarcoma associated herpesvirus (KSHV) microRNAs (miRNAs). MicroRNAs are a class of recently discovered ~22 nucleotide long non-coding RNAs that posttranscriptionally repress mRNAs bearing imperfect matches to the miRNA, primarily in their 3'UTR. KSHV expresses 12 viral miRNAs during latent infection. My immediate research goal is to gain a detailed knowledge of KSHV miRNA targets and to elucidate a possible protective role of these miRNAs against host cell innate immune responses including apoptosis and cell cycle arrest. An extensive set of candidate targets for KSVH miRNA-mediated regulation will be identified by combining gene expression analysis of B cell lines stably expressing physiological levels of one or multiple KSHV miRNAs with binding energy-based target prediction. Interference of the KSHV miRNAs with the induction of apoptosis and cell cycle arrest will be studied (a) based on highly promising candidate targets with known functions in these pathways, (b) through systematic functional interrogation of B cell lines or endothelial cells which ectopically express Individual or multiple KSHV miRNAs, (c) through systematic functional interrogation of latently KSHV Infected primary effusion lymphoma (PEL) cells in which the function of individual or multiple miRNAs has been blocked using antagomirs or sponges. During the mentored phase, I intend to acquire skills in bioinformatics, by taking classes offered at Duke. I will monitor my further development into an independent researcher through regular meetings with my mentor Bryan Cullen and a mentorship committee of experts in computational biology, herpes virology and apoptosis/cell cycle. Furthermore, I expect that the proposed research will both yield sufficient interesting data to apply for an R0l grant early in the independent phase and also further define my longer term research program. RELEVANCE: Relevance: The turnorigenic human gamma herpes viruses KSHV expresses 12 viral mIRNAs during latency. Elucidating functions of KSHV miRNAs is expected to strongly enhance our understanding of KSHV biology and may yield vital clues to the mechanisms of oncogenesis by KSHV and miR-155, an ortholog of the KSHV microRNA miR-K12-11.