Intrinsic and innate barriers to respiratory virus infections

 描述（由申请人提供）：本独立途径提案的主要研究目标是剖析和表征呼吸道病毒感染的内在和先天屏障。虽然呼吸道病毒造成了重大的健康负担，但缺乏有效的抗病毒策略。了解个体宿主因子抑制这些病毒感染的机制可能会导致新的抗病毒策略。最近发现的几种干扰素刺激的基因（ISG），抑制病毒在其感染周期的后期阶段（蝰蛇蛋白-出芽;拴系蛋白-脱离;纤溶酶原激活物抑制剂1 -成熟）验证了这些阶段作为有吸引力的抗病毒靶点。在这个K99奖项的指导阶段，候选人将接受下一代转录组分析和生物信息学的培训，以确定ELF 1（一种ISG和转录因子，赋予广泛的抗病毒抗性）提出的迟效抗病毒程序。此外，基于她先前对甲型流感病毒的迟效抑制剂的鉴定，她将筛选针对其他呼吸道病毒组的抗病毒ISG活性，所述其他呼吸道病毒组由流感B病毒、人副流感病毒、呼吸道合胞病毒、人偏肺病毒、人鼻病毒和人腺病毒组成。最后，候选人将接受CRISPR/Cas9介导的胚胎干细胞（ESC）基因组编辑的培训，并建立将极化的人类气道上皮细胞培养物与肺祖细胞区分开来的方案。这些培养物将用于验证常规组织中的发现 在一个更生理相关的模型系统中培养。候选人全面职业发展计划的其他组成部分是道德操守和赠款撰写方面的课程和研讨会。培训阶段将在洛克菲勒大学（RU）的Charles Rice博士实验室进行，该实验室是世界领先的病毒学研究实验室之一。除了该实验室的重要资源和基础科学专业知识外，候选人还将受益于RU充满活力的研究社区。职业发展的一个关键组成部分将是一个经验丰富的咨询委员会的密切顾问，该委员会由查尔斯·赖斯博士，让-洛朗·卡萨诺瓦博士（均为RU）和阿道夫·加西亚-萨斯特雷博士（西奈山医学院）组成。在K99培训阶段获得的创新技能和有意义的数据集将为ELF 1和独立阶段其他选定命中的详细机制研究奠定基础，并将成为在这个竞争激烈的研究领域中成功的年轻独立研究者的关键。总之，培训将实现候选人的短期目标，即为她的投资组合增加新技术，技能和经验，以及她的长期目标，成为一名独立的研究人员，研究重点是先天免疫系统背景下的病毒-宿主相互作用。

 DESCRIPTION (provided by applicant): The primary research goal of this Pathway to Independence proposal is to dissect and characterize intrinsic and innate barriers to respiratory virus infections. Although respiratory viruses pose a major health burden, there is a paucity of effective, available antiviral strategies. Understanding the mechanisms by which individual host factors inhibit these viral infections may lead to new antiviral strategies. The recent discovery o several interferon-stimulated genes (ISGs) that inhibit viruses at late stages of their infectious cycles (viperin - budding; tetherin - detachment; plasminogen activator inhibitor 1 - maturation) validated such stages as attractive antiviral targets. In the mentored phase of this K99 award, the candidate will be trained in next-generation transcriptome profiling and bioinformatics to identify the late-acting antiviral program raised by ELF1, an ISG and transcription factor conferring broad antiviral resistance. In addition, based on her previous identification of late-acting inhibitors of influenza A virus, she will screen for antiviral ISG activity against a panel f other respiratory viruses, consisting of influenza B virus, human parainfluenzavirus, respiratory syncytial virus, human metapneumovirus, human rhinovirus and human adenovirus. Finally, the candidate will receive training on CRISPR/Cas9-mediated genome editing of embryonic stem cells (ESCs), and establish protocols to differentiate polarized human airway epithelium cultures from lung progenitor cells. These cultures will serve to validate findings from conventional tissue culture in a more physiologically relevant model system. Additional components of the candidate's comprehensive career development plan are courses and seminars in ethics and grant writing. The training phase will be carried out in the laboratory of Dr. Charles Rice at The Rockefeller University (RU), one of the world's leading laboratories in virology research. In addition to the significant resources and basic science expertise in this lab, the candidate will also benefit from RU's vibrant research community. A critical component of career development will be the close counsel of a highly experienced Advisory Committee, composed of Dr. Charles Rice, Dr. Jean-Laurent Casanova (both RU), and Dr. Adolfo Garcia-Sastre (Mt Sinai School of Medicine). The innovative skills and meaningful data sets obtained in the K99 training phase will set the stage for detailed mechanistic studies on ELF1 and other selected hits in the independent phase, and will be key to succeed as a young independent investigator in this highly competitive field of research. In all, the training will fulfill both the candidate's short-erm goals of adding new technologies, skills, and experience to her portfolio, and her long-term goals, to become an independent investigator with a research focus on the virus-host interplay in the context of the innate immune system.