Overcoming host Genetic Redundancy and Pathogen Subversion to Define new host-viral Interfaces

克服宿主遗传冗余和病原体颠覆，定义新的宿主-病毒界面

• 批准号: 10274737
• 负责人: Robert C. Orchard
• 金额: $ 40.51万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-18 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10274737
• 关键词: Antiviral Agents; Area; Attenuated; Binding; Biochemical Genetics; Biology; Bypass; Cell physiology; Communicable Diseases; Complex; Complication; Conflict (Psychology); Epigenetic Process; Family; Genes; Genetic; Genetic Screening; Genome; Histone Acetylation; Host Defense; Immune Evasion; Infection; Influenza; Integration Host Factors; Knowledge; Ligands; Maps; Molecular; Mus; Mutation; Norovirus; Orphan; Pathway interactions; Physiological; Protein Family; Proteins; System; Viral; Virulent; Virus; Virus Replication; comparative; gain of function; insight; pathogen; programs; protein function; receptor; screening; tool

Project Summary/Abstract Viruses have evolved sophisticated mechanisms to hijack host cellular machinery. For their part, hosts have developed their own intricate defense systems. Defining how these opposing strategies have co-evolved increases our understanding of infectious diseases and provides new opportunities to discover unexplored areas of biology. Our understanding of host cellular defense systems has been limited due to genetic redundancy in host genomes. A further complication is that successful pathogens are able to inactivate host antiviral networks during infection. We have recently established a new genetic screening platform that overcomes these roadblocks. First, we bypass genetic redundancy in host genomes through a gain of function screen that identifies antiviral genes. Second, we use both virulent and attenuated strains of viruses in our screening pipeline. Importantly, these attenuated viruses have deletions or mutations in critical, yet poorly understood, immune evasion proteins that antagonize host antiviral restriction factors. We predict that the avirulent strains of these viruses will become sensitive to restriction factors that have otherwise defied molecular identification. Using this strategy, one project will be to identify ancient antiviral genes that reveal unappreciated areas of host- pathogen conflict. For example, we are unmasking how the influenza immune evasion protein, NS1 orchestrates a complex, multifaceted rewiring of host antiviral networks using comparative screens with wild-type and NS1-deficient viruses. We are extending these studies to other virulent-attenuated virus pairings as well. Another project takes newly identified antiviral molecules that inhibit disparate virus families and define the molecular mechanism underlying viral inhibition. Here, we focus on the JADE family of proteins that regulate histone acetylation. We aim to uncover functional redundancy in the JADE family and to determine if it assembles a concerted antiviral epigenetic program. Another project is to leverage newly identified host-pathogen conflicts to discover the physiological functions of host- proteins that are poorly understood. In this regard, we are focusing on the CD300 family of receptor proteins. CD300 genes are undergoing rapid positive selection, yet their physiological function remains largely unexplored. We previously demonstrated that CD300lf is a protein receptor for murine norovirus. We will employ the biochemical, genetic, and cellular tools we developed for studying norovirus-CD300 interactions to define the physiological ligands of CD300 receptors. Determining the ligands for these orphan receptors will provide functional insight into these rapidly evolving proteins. Taken together, we plot a road map for discovering new host-pathogen interfaces, defining their molecular interactions during host defense, and how this relates to the physiological functions of these proteins and pathways.

项目摘要/摘要 病毒已将复杂的机制进化为劫持宿主蜂窝机械。对于他们来说， 主持人已经开发了自己的复杂防御系统。定义这些反对策略如何 共同发展会增加我们对传染病的理解，并为 发现未开发的生物学领域。我们对宿主蜂窝防御系统的理解已经 由于宿主基因组中的遗传冗余而受到限制。另一个并发症是成功的病原体 能够在感染过程中灭活宿主抗病毒网络。我们最近建立了一个新的 克服这些障碍的基因筛选平台。首先，我们绕过遗传冗余 通过鉴定抗病毒基因的功能屏幕的增益，宿主基因组。第二，我们两者都使用 在我们的筛查管道中，有毒的病毒菌株。重要的是，这些减弱了 病毒在关键但知之甚少的免疫逃避蛋白中有缺失或突变， 对抗宿主抗病毒药限制因子。我们预测这些病毒的无毒菌株将 对限制因素敏感，限制因素，这些因素原本是分子鉴定。使用此 策略，一个项目将是确定揭示宿主未见地区的古代抗病毒基因 - 病原体冲突。例如，我们揭露了流感免疫逃避蛋白NS1如何 使用与比较的筛选，策划了宿主抗病毒网络的复杂，多方面的重新布线 野生型和NS1缺陷病毒。我们将这些研究扩展到其他有毒的病毒 配对也是。另一个项目采用抑制不同病毒的新鉴定的抗病毒分子 家族并定义病毒抑制的基本机制。在这里，我们专注于玉 调节组蛋白乙酰化的蛋白质家族。我们旨在发现功能冗余 翡翠家族并确定它是否组装了一致的抗病毒表观遗传学计划。另一个项目 是要利用新确定的宿主 - 病原体冲突来发现宿主的生理功能 - 鲜为人知的蛋白质。在这方面，我们专注于CD300受体家族 蛋白质。 CD300基因正在接受快速阳性选择，但其生理功能仍然存在 在很大程度上没有探索。我们先前证明CD300LF是诺如病毒鼠的蛋白质受体。 我们将使用我们开发的生化，遗传和细胞工具来研究Norovirus-CD300 相互作用以定义CD300受体的生理配体。确定这些配体 孤儿受体将为这些快速发展的蛋白质提供功能洞察力。总的来说，我们 绘制发现新的宿主 - 病原体界面的路线图，定义其分子相互作用 在宿主防御期间，以及这与这些蛋白质和途径的生理功能有何关系。