A ratiometric imaging platform to advance COVID-19 counter measures

新冠肺炎是由SARS冠状病毒2型感染肺上皮细胞引起的一种严重急性呼吸综合征。冠状病毒复制需要一个称为溶酶体的细胞隔间的酸性环境。因此，监测溶酶体积累可能是研究病毒复制和监测病毒载量的一种手段。我们建议重新利用我们在研究神经退行性疾病溶酶体时建立的大量专业知识和行业合作伙伴关系，以开发、优化和验证一种快速、低成本、高通量的方法来测量冠状病毒在肺细胞溶酶体中的积累和复制。因此，该项目的主要目标是，1)开发和优化基于显微镜的溶酶体中SARS-CoV-2水平的测量2)为广泛使用的显微镜软件开发一个模块，该模块将能够快速、准确地测量SARS-CoV-2复制，以及3)验证这一新工具允许研究CoV复制。为此，我们与Quorum Technologies合作，这是一家位于安大略省圭尔夫的加拿大独资企业，开发显微镜和显微镜-生命科学软件。我们开发的比率成像软件模块还将用于进行原则证明测试，以确定系统是否准确地测量SARS-CoV-2复制。鉴于对新冠肺炎患者病毒载量的诊断和预测措施非常缺乏，而且迫切需要，拟议的工作是朝着加快应对新冠肺炎的方向迈出的重要一步。

COVID-19 is a severe acute respiratory syndrome (SARS) caused by infection of lung epithelial cells with the SARS coronavirus 2 (SARS-CoV-2). CoV replication requires the acidic environment of a cell compartment called the lysosome. As such, monitoring the lysosome accumulation may represent a means of studying viral replication and monitoring viral load. We propose to repurpose our considerable expertise and industry partnerships that were developed while studying lysosomes in neurodegenerative disease to develop, optimize and validate a fast, low-cost, high-throughput approach to measuring CoV accumulation and replication in the lysosomes of lung cells. The primary goals of this project are therefore, 1) to develop and optimize a microscopy-based measurement of SARS-CoV-2 levels in the lysosome 2) to develop a module for a widely used microscopy software that will enable fast, accurate measurement of SARS-CoV-2 replication, and 3) to validate that this new tool permits the study of CoV replication. To do this, we have partnered with Quorum Technologies, a Canadian owned enterprise located in Guelph, Ontario that develops microscopes and microscopy-software for the life sciences. The ratiometric imaging software module we develop will also be used to conduct proof-of-principle tests that will determine whether the system accurately measures SARS-CoV-2 replication. Given diagnostic and prognostic measures of viral load in COVID-19 patients are sorely lacking and urgently needed, the proposed work represents an important step towards accelerating COVID-19 countermeasures.