Characterization of novel genes encoded ty RNA and DNA viruses

RNA 和 DNA 病毒编码新基因的表征

基本信息

批准号：
8582320
负责人：
Ralph S Baric
金额：
$ 202.76万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-21 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8582320
关键词：
Acute Address Adult Respiratory Distress Syndrome Agonist Animal Model Antiviral Agents Antiviral Response Apoptosis Apoptotic Attenuated Attenuated Live Virus Vaccine Biochemical Biochemistry Bioinformatics Biological Assay Biology Candidate Disease Gene Categories Cell Culture Techniques Cells Chiroptera Chronic Confocal Microscopy Coronavirus Coronavirus Infections DNA Viruses Defense Mechanisms Development Disease Disease Outbreaks Disease Outcome Disease Progression Ebola virus Epidemic Family Functional RNA Future Gene Expression Gene Targeting Genes Genome Goals Grant Growth Herpesviridae Human Human Herpesvirus 8 Human Virus Immune Immune response Immunity Immunology procedure In Vitro Infection Inflammation Influenza A Virus, H1N1 Subtype Influenza A Virus, H5N1 Subtype Laboratories Leadership Length Life Location Lower respiratory tract structure Lung Mediating Modeling Molecular Cloning National Institute of Allergy and Infectious Disease Natural Immunity Nucleotides Open Reading Frames Pathogenesis Pathway interactions Play Population RNA RNA Viruses Reagent Resources Role Severity of illness Signal Transduction Study models Synthetic Genes Systemic infection Testing Therapeutic Intervention Untranslated RNA Vaccine Design Viral Viral Genes Viral Pathogenesis Viral Proteins Virus Virus Diseases Virus Replication Work aged base data management defense response design disorder control expression vector gene function genetic evolution global health high throughput screening human coronavirus human disease improved in vivo influenzavirus latent infection loss of function microbial mortality mutant new therapeutic target novel pathogen positional cloning pressure programs public health relevance recombinant virus respiratory response screening therapeutic target trait viral DNA virus genetics virus host interaction virus pathogenesis

项目摘要

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.

描述（由申请人提供）：病毒编码未表征基因的新子集（预测和假设的 ORF 和非编码 RNA），这些基因可以表达以在体外和体内调节病毒复制效率和/或宿主抗病毒反应。利用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，我们测试了这样的假设：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，从而确定严重的疾病结果。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组明确的生化和免疫学测定来识别、表征并确定未表征基因在疾病中的作用。肺部（例如，H5N1、SARS-CoV 和人类冠状病毒 EMC-1）和全身感染（例如，埃博拉和人类疱疹病毒 8），无论是在体外还是在某些情况下，在体内。具体来说，我们测试了以下假设：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、先天免疫感应、炎症小体、细胞凋亡和/或 NF-κB 信号传导。为了实现这些目标，RNA和DNA病毒发病机制和免疫方面高度互动的专家小组共同创建了一个强大的筛选平台，可以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。通过识别不同病毒病原体针对的共同关键宿主瓶颈基因，我们确定了改善体内疾病结果的广泛相关治疗靶点的基本原理。重要的是，该平台：a）便携式，b）可以快速应用于其他高致病性呼吸道和微生物病原体，c）将快速识别治疗干预的新靶标，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爆发的反应。