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.

项目总结/摘要 病毒已经进化出复杂的机制来劫持宿主的细胞机器。对他们来说， 宿主已经发展出自己复杂的防御系统。定义这些对立的策略 共同进化增加了我们对传染病的理解，并提供了新的机会， 发现生物学中未被探索的领域。我们对宿主细胞防御系统的理解 由于宿主基因组中的遗传冗余而受到限制。更复杂的是，成功的病原体 能够在感染期间破坏抗病毒网络。我们最近建立了新的 克服这些障碍的基因筛选平台。首先，我们绕过基因冗余， 宿主基因组，通过功能筛选，确定抗病毒基因的增益。第二，我们两者都使用 强毒和减毒病毒株在我们的筛选管道。重要的是，这些衰减 病毒在关键的但知之甚少的免疫逃避蛋白中有缺失或突变， 拮抗宿主抗病毒限制因子。我们预测，这些病毒的无毒株将 变得对限制性因子敏感，否则这些限制性因子无法进行分子鉴定。使用此 战略，一个项目将是确定古老的抗病毒基因，揭示不受重视的领域的主机- 病原体冲突例如，我们正在揭示流感免疫逃避蛋白NS 1 使用比较筛选，协调宿主抗病毒网络的复杂，多方面的重新布线， 野生型和NS 1缺陷型病毒。我们正在将这些研究扩展到其他毒性减毒病毒 pairs也是。另一个项目采用新发现的抗病毒分子， 家族并定义了病毒抑制的分子机制。在这里，我们专注于玉 调节组蛋白乙酰化的蛋白质家族。我们的目标是发现 JADE家族，并确定它是否组装了一个协调一致的抗病毒表观遗传程序。另一个项目 是利用新发现的宿主-病原体冲突来发现宿主的生理功能， 我们对这些蛋白质知之甚少。在这方面，我们关注的是CD 300受体家族， proteins. CD 300基因正在经历快速正向选择，但其生理功能仍然存在 大部分未开发。我们以前证明，CD 300 lf是一个蛋白受体的小鼠诺如病毒。 我们将使用我们为研究诺如病毒-CD 300而开发的生物化学、遗传学和细胞学工具 这些相互作用以定义CD 300受体的生理配体。确定这些的配体 孤儿受体将提供对这些快速进化的蛋白质的功能性洞察。总之，我们 为发现新的宿主-病原体界面绘制路线图，定义它们的分子相互作用 在宿主防御过程中，以及这与这些蛋白质和途径的生理功能的关系。