New methods and tools to improve reproducibility and utility of 3-D organoids as an in vitro cell culture model of viral infection

人类与致病病毒之间的相互作用通常包括这些病毒与全身受感染器官的多种宿主细胞类型之间复杂而独特的相互作用。具有感染性的病毒颗粒对靶器官的识别是病毒感染结局的重要决定因素。就新型蚊媒病毒，如寨卡病毒(ZIKV)和西尼罗河病毒(WNV)而言，它们感染脑细胞(嗜神经病毒)的能力与包括脑损伤在内的危及生命的脑部感染有关。随着与这些新出现的嗜神经性病毒相关的全球健康问题的增加，迫切需要适当的分子工具来了解它们在目标器官中的生命周期，为开发治疗性医疗设备铺平道路。**在这项为期1年的NSERC CRD赠款中，不列颠哥伦比亚大学(UBC)的Jean博士的团队与加拿大干细胞技术公司的科学家Leon Chew合作，正在提议开发和优化两个多细胞3D迷你大脑模型(“脑器官”)，以在培养皿中复制ZIKV生命周期和3D细胞培养系统中具有感染性的病毒颗粒的强大生产。**干细胞技术的科学家最近开发了从人类干细胞制造3D迷你大脑的协议。然而，开发和优化ZIKV感染的3D迷你大脑模型需要开发有效的有机物感染方案和安全的ZIKV处理程序。Jean博士的UBC团队成员具有Jean博士在分子病毒学方面的科学专长，并且他的团队成员可以使用UBC的世界级基础设施和最先进的设备，这将为这一与干细胞技术公司的合作研究项目提供支持。该项目将促进标准化工作流程程序和安全操作程序的发展，这些程序对于在加拿大和世界各地应用基于“迷你大脑”的有机化合物技术的病毒学研究至关重要。****

Interactions between humans and pathogenic viruses often include an intricate and unique interplay between these viruses and multiple host cell types of the infected organs throughout the body. The identity of the target organ by the infectious virus particles is an important determinant of viral infection outcomes. In the case of emerging mosquito-borne viruses such as Zika virus (ZIKV) and West Nile virus (WNV), their ability to infect brain cells (neurotropic viruses) is associated with life-threatening brain infections including brain damage. With the increase of global health concerns associated with these emerging neurotropic viruses, there is an urgent need for appropriate molecular tools to understand their lifecycle in the target organ to pave the way for developing a therapeutic armamentarium.**In this 1-year NSERC CRD grant, Dr. Jean's team at the University of British Columbia (UBC) in collaboration with Leon Chew, scientist at STEMCELL Technologies (Canada), are proposing to develop and optimize two multicellular 3D mini-brain models ("brain organoids") to reproduce in petri dishes the ZIKV lifecycle and the robust production of infectious virus particles in 3D cell culture systems.**Scientists at STEMCELL Technology have recently developed protocols to produce 3D mini-brains from human stem cells. However, developing and optimizing 3D mini-brain models for ZIKV infection require the development of effective organoid infection protocols and safe handling procedures for ZIKV. Dr. Jean's UBC team members are well positioned to succeed in this project with Dr. Jean's scientific expertise in molecular virology and the access by his team members to world-class infrastructures and state-of-the-art equipment at UBC, which will support this collaborative research project with STEMCELL Technologies. This project will catalyze the development of standardized workflow procedures and safety operational procedures that are critical for applying "mini-brain" organoid-based technology in virology research in Canada and around the world. ****