Characterization of novel genes encoded ty RNA and DNA viruses

描述（由申请人提供）：病毒编码新的未表征基因亚群（预测的和假设的orf和非编码rna），这些基因可以在体外和体内表达以调节病毒复制效率和/或宿主抗病毒反应。利用可导致急性和慢性危及生命的疾病结果的高致病性人类呼吸道和全身病毒，我们验证了RNA和DNA病毒编码共同和独特的机制来操纵病毒复制效率和宿主反应以决定严重疾病结果的假设。为了解决这一假设，该提案利用新型表达载体平台、合成基因设计、反向遗传学、人类疾病动物模型以及一套明确的生化和免疫学分析，在体外和某些情况下在体内识别、表征并确定未表征基因在肺部（例如H5N1、SARS-CoV和人类冠状病毒EMC-1）和全身感染（例如埃博拉病毒和人类疱疹病毒8）中的作用。具体来说，我们验证了这样一个假设，即这些病毒未表征的基因可能具有自动调节病毒复制效率的功能，和/或作为激动剂或拮抗宿主细胞内环境以增强病毒复制的功能，最有可能改变p53、先天免疫感知、炎性体、细胞凋亡和/或NF-？B信号。为了实现这些目标，一个高度互动的RNA和DNA病毒发病机制和免疫专家小组共同努力创建一个强大的筛选平台，快速识别和表征这些未表征的基因在复制和发病机制中的功能。通过鉴定被不同病毒病原体靶向的常见关键宿主瓶颈基因，我们确定了改善体内疾病结果的基本原理和广泛相关的治疗靶点。重要的是，该平台是：a)便携，b)可快速应用于其他高致病性呼吸道和微生物病原体，c)将快速确定治疗干预的新靶点，d)改进减毒活疫苗或载体病毒疫苗设计策略，以及e)改善全球对新发现的人群流行病暴发的反应。

DESCRIPTION (provided by applicant): Viruses encode novel subsets of uncharacterized genes (predicted and hypothetical ORFs and noncoding RNAs) which can be expressed to modulate virus replication efficiency and/or host antiviral responses both in vitro and in vivo. Using highly pathogenic human respiratory and systemic viruses which cause acute and chronic life-threatening disease outcomes, we test the hypothesis that RNA and DNA viruses encode common and unique mechanisms to manipulate virus replication efficiency and host responses to determine severe disease outcomes. To address this hypothesis, the proposal takes advantage of novel expression vector platforms, synthetic gene design, reverse genetics, animal models of human disease, and a defined set of biochemical and immunologic assays to identify, characterize and then determine the role of uncharacterized genes in the lung (e.g., H5N1, SARS-CoV and human coronavirus EMC-1) and in systemic infections (e.g., Ebola and Human Herpes virus 8) both in vitro and in some instances, in vivo. Specifically, we test the hypothesis that these viral uncharacterized genes may function to auto-regulate virus replication efficiency, and/or function as an agonist or antagonize the host intracellular milieu to enhance virus replication, most likely be altering p53, innate immune sensing, inflammasome, apoptosis, and/or NF-?B signaling. To achieve these goals, a highly interactive group of experts in RNA and DNA virus pathogenesis and immunity work collectively to create a robust screening platform that rapidly identifies and characterizes the function of these uncharacterized genes in replication and pathogenesis. By identifying common key host bottleneck genes that are targeted by disparate virus pathogens, we identify rationale broadly relevant therapeutic targets for ameliorating disease outcomes in vivo. Importantly, this platform is: a) portable, b) can be rapidly applied to other highly pathogenic respiratory and microbial pathogens, c) will rapidly identify novel targets for therapeutic intervention, d) improve strategies for live attenuated or vectored virus vaccine design, and e) improve global responses to newly identified, epidemic disease outbreaks in human populations.