3D multi-target imaging of RNA virus replication deep in cells at the nanoscale

这项合作旨在为开发和应用尖端超分辨率成像技术来研究RNA病毒与宿主的相互作用奠定基础。这种合作将使加拿大人受益，因为它直接贡献了新的成像技术，获得了关于基本RNA病毒生物学的新知识，并间接有助于开发有效的干预战略来抗击RNA病毒感染。由核糖核酸病毒SARS-CoV-2引发的新冠肺炎大流行迄今已导致40,000多人死亡，国内生产总值下降18%。RNA病毒感染还会引起牲畜疾病和经济损失。自21世纪初以来，加拿大爆发的牛海绵状脑病造成了63亿美元的损失，2004年不列颠哥伦比亚省爆发的禽流感导致了高达3.8亿美元的经济损失。一种阳性的RNA病毒，诺沃克病毒，是食源性疾病的主要原因。为了制定诊断和干预策略，我们必须了解病毒在宿主中的生物活性。RNA病毒依靠宿主生存和复制。与其他阳性RNA病毒一样，诺如病毒(NOV)感染可引起宿主细胞内细胞膜的重塑，以形成病毒复制复合体(Rc)。在RC中发生了NOV复制，以及病毒组装和输出。RC保护病毒基因组免受宿主的先天免疫。挑战在于，目前的技术不允许在活体标本中直接可视化RC形成的动力学。2014年诺贝尔奖获得者超分辨率成像技术打破了光的衍射极限，允许在病毒感染的活细胞中分析微小的RC结构和单分子水平上详细的病毒与宿主的相互作用。这项合作将促进先进的生物成像技术在研究NOV RC的形成和结构方面的应用。新技术发现的关于RNA病毒复制和生物发生的新知识将为未来疫情的应对计划提供信息。

The collaboration aims to lay the ground for the development and application of cutting-edge super-resolution imaging technology to study RNA virus-host interactions. This collaboration will have benefits for Canadians by directly contributing new imaging technology for new knowledge on the fundamental RNA virus biology and by indirectly contributing to the development of effective intervention strategies to battle RNA virus infection. The COVID-19 pandemic caused by an RNA virus, SARS-CoV-2, has resulted in over 40,000 deaths and an 18% GDP decline to date. RNA virus infection also causes livestock disease and economic loss. Canadian outbreaks of bovine spongiform encephalopathy cost $6.3 billion since the early 2000s and the 2004 outbreak of avian influenza in BC led to economic losses of up to $380 million. A positive RNA virus, norovirus, is a leading cause of foodborne diseases. To develop diagnostic and intervention strategies, we must understand the biological activity of the virus in its host. RNA virus relies on its host for survival and replication. Like other positive RNA viruses, norovirus (NoV) infection induces the remodeling of the host cell's intracellular membranes for the formation of viral replication complexes (RC, <300nm). NoV replication occurs in the RC, as well as virus assembly and export. The RC protects the viral genome from the host's innate immunity. The challenge is that the current technology does not allow direct visualization of the dynamics in the formation of the RC in the live specimen. The 2014 Nobel Prize-winning super-resolution imaging technology breaks the diffraction limit of light and allows the analysis of tiny RC structures and the detailed virus-host interactions on a single-molecule level in virus-infected live cells. This collaboration will foster the application of advanced bioimaging technology in studying the formation and structure of the NoV RC. The new knowledge on RNA virus replication and biogenesis uncovered by the new technology will inform the response plan for future outbreaks.