Matt Buechner
$200,000
University of Rochester
New York
Biological Sciences (BIO)
SARS-CoV-2 is the causative agent of the COVID-19 pandemic, a devastating infectious disease that in only few months has caused a global health and economic crisis. Similar to the SARS and MERS epidemics, COVID-19 originated from a coronavirus that was transmitted from bats to humans. All these epidemics evidence that spillovers of this type of viruses are frequent and future transmissions will inevitably occur. Therefore, there is a critical need to understand the mechanism by which coronaviruses in general, and SARS-CoV-2 in particular, move from its natural host to humans. This project will address this critical research question using biochemical and microscopy methods. This research also aims at training the next generation of scientists by providing them with the necessary tools and mentoring skills to carry out cutting-edge research with high impact to our society: from deciphering how this class of viruses disseminate at the molecular level to gaining a deeper understanding of the human and animal behavioral interactions that have resulted in the emergence of this disease. This project will investigate the role of the autophagy machinery in SARS-CoV-2 dissemination in different hosts. Autophagy is a highly conserved survival pathway that remodels cellular membranes to trap waste materials for their elimination. This research will test the innovative hypothesis that SARS-CoV-2 usurps the cellular autophagy machinery with the goal of creating two distinct structures: membrane scaffolds for the multiplication of the virus genome (viroplasms) and antidefense shields to conceal virus materials that would otherwise be detected by the immune response (i.e. dsRNA intermediates). This project has two goals to test this hypothesis:(1)Characterizing the membrane structures generated during SARS-CoV-2 infection in bat, ferret and human cells. Lipidomic and proteomic studies will be performed to reveal the biochemical makeup of the membrane scaffolds and antidefense shields generated by SARS-CoV-2 in different hosts. (2) Identifying the molecular mechanisms by which SARS-CoV-2 intersects with the autophagy machinery. The goal of this aim is to monitor the effects of SARS-CoV-2 on key checkpoints of autophagy. Confocal microscopy, flow cytometry and biochemical assays will be used to measure autophagosome formation, autophagy flux and the molecular composition of the checkpoints that regulate autophagy in bat, ferret and human cells.This RAPID award is made by the Cellular Dynamics and Function Program in the Division of Molecular and Cellular Biosciences, using funds from the Coronavirus Aid, Relief, and Economic Security (CARES) Act.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.