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

描述(由申请人提供)：2010年夏天，1 AM希望转到一个独立的教员职位，从事卡波西肉瘤相关疱疹病毒(KSHV)microRNAs(MiRNAs)的功能分析。MicroRNAs是最近发现的一类长约22个核苷酸的非编码RNA，它在转录后抑制与miRNA不完全匹配的mRNAs，主要是在其3‘非编码区。KSHV在潜伏感染过程中表达12个病毒miRNAs。我目前的研究目标是获得KSHV miRNA靶标的详细知识，并阐明这些miRNAs对宿主细胞先天性免疫反应(包括细胞凋亡和细胞周期停滞)可能的保护作用。通过结合稳定表达一个或多个KSHV miRNAs生理水平的B细胞系的基因表达分析和基于结合能量的靶点预测，将识别一组广泛的KSVH miRNA介导的调控候选靶点。KSHV miRNAs对诱导细胞凋亡和细胞周期停滞的干扰将被研究：(A)基于在这些途径中具有已知功能的极有希望的候选靶点，(B)通过对异位表达单个或多个KSHV miRNAs的B细胞系或内皮细胞的系统功能询问，(C)通过对潜伏感染KSHV的原发渗出性淋巴瘤(PEL)细胞的系统功能询问，在PEL细胞中，单个或多个miRNAs的功能已被抗原虫或海绵阻断。在指导阶段，我打算通过参加杜克大学提供的课程来获得生物信息学方面的技能。我将定期与我的导师布莱恩·卡伦和一个由计算生物学、疱疹病毒学和细胞凋亡/细胞周期方面的专家组成的导师委员会会面，以监督我进一步发展成为一名独立研究员。此外，我预计拟议的研究将产生足够的有趣数据，以便在独立阶段早期申请R01拨款，并进一步确定我的长期研究计划。 相关性：致转化性人类伽马疱疹病毒KSHV在潜伏期表达12种病毒miRNAs。阐明KSHV miRNAs的功能有望大大加强我们对KSHV生物学的理解，并可能为KSHV和KSHV microRNA miR-K12-11的同源基因miR-155的致癌机制提供重要线索。

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.