A Secreted Viral Protein Determines Norovirus Persistence By Immune Evasion

项目摘要/摘要 这份提案描述了Sanghyun博士的四年职业发展计划和研究战略 Lee将从博士后过渡到研究病毒宿主的独立学术教员职位 互动。该提案的总体研究目标是阐明一种分泌的病毒 非结构蛋白拮抗宿主免疫，并评价该病毒因子作为新的疫苗靶点。 应聘者：我的科学生涯主要研究病毒和宿主在这两个分子上的相互作用 和生理水平。在我与韩国首尔国立大学安光熙博士的博士合作中，我 致力于了解人类巨细胞病毒(HCMV)的毒力，并特别关注 影响人类巨细胞病毒毒力的因素。作为斯基普·维珍博士华盛顿实验室的博士后 大学医学院，我研究了诺沃克病毒的细胞嗜性和关键宿主免疫系统 控制肠道内诺沃克病毒感染的决定因素。 职业发展计划：除了这里概述的研究计划，我将投入大约15% 把我的时间花在训练活动上。我会在我的导师的指导下继续我的职业发展 维珍医生和我的共同导师加亚·阿马拉辛格博士。我已经组建了一个职业咨询委员会 由Michael Diamond博士、Megan Baldridge博士和Haina Shin博士组成，将评估和促进 我的研究进展和向独立的过渡。除了这些培训活动，我还将准备 通过以下途径提高撰写助学金和面试的技能，从而过渡到独立的教员职位 在WUSM举办研讨会，并开始规划新的财务、管理和指导职责 一名独立调查员的身份。 研究项目：人类诺如病毒(HNov)是全球胃肠炎的主要原因。然而， 目前还没有获得批准的疫苗或疗法。通过小鼠诺如病毒(MNoV)模型，我们鉴定了丛状细胞， 一种罕见的肠上皮细胞(IEC)，作为MNoV排泄物和持久性的储存库。遗传 研究表明，病毒非结构蛋白NS1而不是病毒表面蛋白是病毒的决定因素 丛生细胞嗜性，干扰素-λ是一个关键的宿主决定因素。我们的初步数据显示 Ns1蛋白被宿主caspase(S)切割并分泌到细胞外间隙，ns1被阻断 干扰素-λ的产生。因此，我们假设分泌的ns1蛋白能拮抗干扰素-λ的产生。 这种拮抗作用是体内丛状细胞趋向性的关键决定因素。这项研究的长期目标是 了解mnov分泌ns1和免疫逃避干扰素-λ的分子机制，并应用我们的 评估分泌的NS1作为人类疫苗和治疗开发候选者的研究结果。

Project Summary/Abstract This proposal describes a four year career development plan and a research strategy for Dr. Sanghyun Lee to transition from a postdoctoral fellow to an independent academic faculty position investigating virus-host interactions. The overall research goal of the proposal is to elucidate the mechanism by which a secreted viral non-structural protein antagonizes host immunity, and to evaluate this viral factor as a novel vaccine target. Candidate: I have focused my science career on understanding virus and host interactions at both molecular and physiological levels. In my Ph.D. work with Dr. Kwangseog Ahn at Seoul National University in Korea, I worked on understanding the virulence of human cytomegalovirus (HCMV), and specifically focused on the factors contributing to HCMV virulence in human patients. As a postdoc in Dr. Skip Virgin's lab at Washington University School of Medicine, I studied the cellular tropism of norovirus and the key host immune system determinants controlling norovirus infection in the gut. Career Development Plan: In addition to the research proposal outlined here, I will devote approximately 15% of my time to training activities. I will continue my professional development under the guidance of my mentor Dr. Virgin's and my co-mentor Dr. Gaya Amarasinghe. I have assembled a career advisory committee composed of Dr. Michael Diamond, Dr. Megan Baldridge, and Dr. Haina Shin that will evaluate and facilitate my research progress and transition to independence. Beyond these training activities, I will prepare to transition to an independent faculty position by improving grant-writing and interviewing skills through workshops at WUSM, and begin planning for the new financial, management, and mentorship responsibilities of an independent investigator. Research Project: Human noroviruses (HNoVs) are the leading global cause of gastroenteritis. However, there are no approved vaccines or therapeutics. With Murine norovirus (MNoV) model, we identified Tuft cells, a rare type of intestinal epithelial cell (IEC), as the reservoir for MNoV fecal shedding and persistence. Genetic studies revealed that viral non-structural protein NS1, rather than a viral surface protein, is the determinant of Tuft cell tropism, and interferon-lambda (IFN-λ) to be a key host determinant. Our preliminary data suggests that the NS1 protein is cleaved by host caspase(s) and secreted into the extracellular space, and NS1 blocks IFN-λ production. Therefore, we hypothesize that secreted NS1 protein antagonizes IFN-λ production, and that this antagonism is the key determinant of in vivo Tuft cell tropism. The long-term goal of this study is to understand the molecular mechanism of NS1 secretion and IFN-λ immune evasion by MNoV, and to apply our findings to evaluate secreted NS1 as a candidate for vaccine and therapeutic development in humans.