A ratiometric imaging platform to advance COVID‑19 counter measures
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.