Intrinsic and innate barriers to respiratory virus infections

 描述(由申请人提供)：这个独立之路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障碍。尽管呼吸道病毒构成了主要的健康负担，但缺乏有效的、可用的抗病毒策略。了解单个宿主因素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基因(ISGs)可以在病毒感染周期的后期阶段(毒蛇出芽、拴系分离、纤溶酶原激活物抑制物1成熟)抑制病毒，证实了这些阶段是有吸引力的抗病毒靶点。在K99奖项的指导阶段，候选人将接受下一代转录组图谱和生物信息学方面的培训，以确定Elf1提出的晚期作用抗病毒计划，Elf1是一种ISG和转录因子，具有广泛的抗病毒耐药性。此外，根据她先前发现的甲型流感病毒的迟效抑制物，她将筛选针对其他呼吸道病毒的抗病毒ISG活性，这些病毒包括B型流感病毒、人类副流感病毒、呼吸道合胞病毒、人类偏肺病毒、人类鼻病毒和人腺病毒。最后，候选人将接受CRISPR/Cas9介导的胚胎干细胞(ESCs)基因组编辑方面的培训，并建立区分极化的人类呼吸道上皮细胞和肺祖细胞的方案。这些培养物将用于验证传统组织的发现。 在生理上更相关的模式系统中的文化。候选人的全面职业发展计划的其他组成部分是道德和补助金撰写方面的课程和研讨会。培训阶段将在洛克菲勒大学查尔斯·赖斯博士的实验室进行，洛克菲勒大学是世界领先的病毒学研究实验室之一。除了这个实验室的重要资源和基础科学专业知识外，候选人还将从RU充满活力的研究社区中受益。职业发展的一个关键组成部分将是由查尔斯·赖斯博士、Jean-Laurent Casanova博士(均为RU)和Adolfo Garcia-Sastre博士(西奈山医学院)组成的经验丰富的咨询委员会的密切咨询。在K99培训阶段获得的创新技能和有意义的数据集将为独立阶段对Elf1和其他选定的Hit进行详细的机械研究奠定基础，并将是在这个竞争激烈的研究领域作为一名年轻的独立研究人员取得成功的关键。总而言之，培训将实现候选人的短期目标，即在她的投资组合中增加新的技术、技能和经验，以及她的长期目标，即成为一名独立的调查员，专注于研究病毒与宿主在先天性免疫系统中的相互作用。

 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.