GSE210497
SARS-CoV-2 pathogenesis mediated by E protein's PBM was prevented by modulators of CFTR function
The three human deadly coronaviruses (CoVs) (SARS-CoV, MERS-CoV and SARS-CoV-2) have infected around eight thousand, two thousand six hundred and 262 million people so far (December 2021), causing the death of 10%, 37% and 2% of them, respectively, so their implication in health is very important. These CoVs have proteins with a PBM motif that binds to PDZ cell domains. PDZ domains are found in more than 400 cellular proteins, therefore, viruses with PBM motifs have a high potential to modify cell behavior. It was studied the implication of the E protein PBM motif of various virulent or attenuated human CoVs (hCoVs) in the pathogenesis induced by these viruses. Variants of SARS-CoV, MERS-CoV and SARS-CoV-2 that lack the E protein PBM have been generated by reverse genetics and their pathogenicity has been analyzed in mice. The PBM motifs of these three hCoVs have been shown to be virulence factors and participate in their replication. Furthermore, a collection of SARS-CoV mutants has been constructed in which the E protein PBM domain was replaced by the one derived from virulent or attenuated hCoVs, and their virulence was analyzed. A virulence gradient was observed, depending on whether the E protein PBM domain was derived from an attenuated or virulent hCoV. The gene expression patterns associated with the different PBM motifs in lungs of mice infected with SARS-CoV was analyzed by deep sequencing of the mRNAs expressed in lungs of infected mice, and it was observed that the E protein PBM motif of SARS-CoV and SARS-CoV-2 dysregulates the expression of genes related to ion transport and cell homeostasis. Specifically, a decrease of the mRNA expression of the cystic fibrosis transmembrane conductance regulator (CFTR), which is essential for edema resolution, was observed. The reduction of CFTR mRNA levels was associated with edema accumulation in lungs of mice infected with SARS-CoV and SARS-CoV-2. The effect of compounds that modulate the expression and activity of CFTR on the pathogenesis and replication of SARS-CoV-2 was studied and it was observed that these compounds drastically reduced the production of SARS-CoV-2 in cell culture and protect against SARS-CoV-2 pathogenesis. These results showed the high relevance of the PBM motif in the replication and virulence of CoVs, and have allowed the identification of cellular targets for the selection of antivirals. Overall design: Comparative gene expression analysis of RNA-seq data for mouse lungs infected with Wild-type or PBM-modifed coronaviruses ("3a", "GGGG", "Mock", "WT", "OC43", "HKU1", "NL63", "MERS") at two different times post infection (2dpi, 4dpi).Three biological replicates per sample.