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

此次合作旨在为研究RNA病毒与宿主相互作用的尖端超分辨率成像技术的开发和应用奠定基础。这种合作将使加拿大人受益，因为它直接提供新的成像技术，获得关于基本RNA病毒生物学的新知识，并间接促进制定对抗RNA病毒感染的有效干预战略。由RNA病毒SARS-CoV-2引起的COVID-19大流行迄今已导致4万多人死亡，国内生产总值下降18%。RNA病毒感染也会造成家畜疾病和经济损失。自2000年代初以来，加拿大爆发的牛海绵状脑病造成了63亿加元的损失，2004年不列颠哥伦比亚省爆发的禽流感造成了高达3.8亿加元的经济损失。RNA阳性病毒诺如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其宿主中的生物活性。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）感染诱导宿主细胞细胞膜重塑，形成病毒复制复合物（RC, <300nm）。病毒复制发生在RC中，以及病毒组装和导出中。RC保护病毒基因组免受宿主先天免疫的侵害。面临的挑战是，目前的技术不允许直接可视化的动态形成的RC在活的标本。2014年获得诺贝尔奖的超分辨率成像技术突破了光的衍射极限，可以在病毒感染的活细胞的单分子水平上分析微小的RC结构和详细的病毒-宿主相互作用。此次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。新技术所揭示的关于RNA病毒复制和生物发生的新知识将为今后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.