Viral Diversity and Pathogenicity in Mucosal Respiratory and Gastrointestinal Disease

项目总结 人类诺如病毒(HuNoV)是NIAID B类病原体，是全球急性胃肠炎的主要病因， 而呼吸道合胞病毒(RSV)是一种C类病原体，是全球主要的呼吸道病原体。 孩子们。新城疫病毒感染在所有年龄段都导致显著的急性发病率，慢性疾病 免疫功能受损的癌症和移植患者，以及幼儿和老年人的死亡。RSV是 儿童下呼吸道发病率和死亡率的主要原因，并在很大程度上有助于 疾病和死亡中的免疫功能受损，那些患有共病的人和老年人。疾病的负担 这些粘膜病原体的经济影响刺激了疫苗开发的广泛努力。 由于预计在未来5年内针对这两种病原体的疫苗，这个许可前窗口，连同我们的 利用干细胞来源的人肠道成功培养HuNov的研究进展 有机化合物(HIOS)和最近对人类肺器官中RSV感染的描述(HLOS)提供了理想的 在相关人类文化系统中应用分析基因组学和功能研究的窗口，以解决关键问题 关于呼肠孤病毒和呼吸道合胞病毒的多样性、进化和毒力的问题。我们提议的研究谨慎地利用 精选临床样本，涵盖新城疫病毒和呼吸道合胞病毒病的病谱，包括：(I)急性胃肠炎 来自两个人群(美国和香港)的具有免疫能力的儿童，(Ii)#年急性呼吸道感染 疾控中心赞助的新疫苗监测网络中的儿童，(Iii)受控良好的志愿者HuNov 挑战研究，(Iv)慢性新城疫病毒感染或急性呼吸道合胞病毒感染的移植和免疫受损患者 感染，(V)硝唑尼特(NTZ)治疗慢性新城疫的首个随机双盲临床试验 在移植接受者中，以及(Vi)在#年RSV-F疫苗(Novavax)第一阶段III试验中的妇女和婴儿 健康的孕妇预防其婴儿的疾病。除了病原体定向测序外，我们还 将使用高通量、集成测序来评估生态位在病毒致病中的作用 临床标本细菌体和病毒体的图谱分析方法。黄瓜全长(FL)基因组分析 HIE中的HuNov和功能研究将提供关于不同人群中病毒多样性的关键信息 随着时间的推移，在不同的宿主中，存在基因内型和基因间型重组体，以及 抗NTZ处理突变体的出现。呼吸道合胞病毒FL基因组分析及功能研究 HLOS和其他细胞系将建立RSV疾病严重程度的特定病毒特征，使人们能够理解 免疫压力和免疫疗法对病毒基因组包括抗原位点特异性的影响 基序，以及B和T细胞表位的变化。科学界将从大规模的 HuNov和RSV FL基因组数据库，以及在新型人类文化中的功能研究 在疫苗接种前全面解决病毒基因组特征、微生物生态和宿主反应 引言。识别生物标记物、病毒和宿主靶点将指导治疗和治疗学的发展。

PROJECT SUMMARY Human norovirus (HuNoV), a NIAID Category B pathogen, is the leading cause of acute gastroenteritis globally, and respiratory syncytial virus (RSV), a Category C pathogen, is the major global respiratory pathogen of children. HuNoV infections result in significant acute morbidity in all age groups, chronic disease in immunocompromised cancer and transplant patients, and death in young children and older adults. RSV is the leading cause of lower respiratory tract morbidity and mortality among children, and contributes significantly to illness and death in the immunocompromised, those with co-morbidities and older adults. The burden of disease and economic impact of these mucosal pathogens have stimulated extensive efforts for vaccine development. With vaccines anticipated in the next 5 years for both pathogens, this pre-licensure window, together with our recent development of a successful cultivation system for HuNoVs using stem cell-derived human intestinal organoids (HIOs) and the recent description of RSV infections in human lung organoids (HLOs) provide an ideal window to apply analytical genomics and functional studies in relevant human culture systems to address key questions about HuNoV and RSV diversity, evolution and virulence. Our proposed studies utilize carefully selected clinical specimens covering the spectrum of HuNoV and RSV illness, including: (i) acute gastroenteritis in immunocompetent children from two populations (USA and Hong Kong), (ii) acute respiratory infections in children in the CDC sponsored New Vaccine Surveillance Network, (iii) well-controlled volunteer HuNoV challenge studies, (iv) transplant and immunocompromised patients with chronic HuNoV infection or acute RSV infection, (v) the first randomized double-blind clinical trial of nitazoxanide (NTZ) for treatment of chronic HuNoV in transplant recipients, and (vi) women and infants in the first phase III trial of an RSV-F vaccine (Novavax) in healthy pregnant women for preventing disease in their infants. In addition to pathogen-targeted sequencing, we will evaluate the role of the ecological niche in viral pathogenesis using high throughput, integrated sequencing approaches to profile the bacteriome and virome of clinical samples. Full-length (FL) genomic analyses of HuNoV and functional studies in HIEs will provide critical information on virus diversity in different populations and over time in diverse hosts, the presence of intragenotypic and intergenotypic recombinants, and the appearance of mutants with treatment resistance to NTZ. FL genomic analyses of RSV and functional studies in HLOs and other cell lines will establish specific viral signatures of RSV disease severity, enable understanding the effects of immune pressure and immunotherapeutics on the viral genome including antigenic site specific motifs, and changes in B and T cell epitopes. The scientific community will benefit immensely from large databases of FL genomes for HuNoV and RSV, and functional studies in novel human cultures that comprehensively address viral genomic signatures, microbial ecology and host responses before vaccine introduction. Identifying biomarkers, viral and host targets will guide treatment and development of therapeutics.