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SARS-CoV-2 pathogenesis mediated by E protein's PBM was prevented by modulators of CFTR function

ABSTRACT: 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.

ORGANISM(S): Mus musculus

PROVIDER: GSE210497 | GEO | 2023/09/22

REPOSITORIES: GEO

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Project description:PDZ (Post-synaptic density, PSD-95)/ Disc large, Dlg/ Zonula occludens, ZO-1) proteins are central in the assembly of multiprotein signalling complexes that are formed in distinctive, specialized cell regions. In these complexes different signals are orchestrated and traduced into quick and efficient responses controlling cell polarization and cell communication. Viral pathogens target host PDZ proteins by expressing viral proteins containing a PDZ binding motif (PBM). SARS-CoV-1 and -2 harbour the protein E, a viroporin with a conserved PBM in its carboxyl-terminal region. SARS-CoV-1 E protein is a pathogenicity factor that in epithelia interacts with the PDZ protein PALS1, which promote disruption of cell junctions. SARS-CoV-2 infects epithelia, but also cells from the innate immune response, including monocytes, dendritic cells, and alveolar macrophages. Identification of targets of SARS-CoV-2 E protein in immune cells might offer valuable clues to understand how SARS-CoV-2 alters immune response. Here we generated a SARS-CoV-2 E protein fused to a GFP-tag at the amino terminal. This recombinant protein was used as bait to identify associated proteins in THP-1 cells, a human monocytic cell line that can be differentiated to macrophages and dendritic cells. Analysis of the GFP-E protein interactome provided 372 proteins that fall into different functional groups. Only eight of these proteins harbor PDZ domains, including the cell polarity protein ZO-2 and the chemoattractant IL-16. Syntenin, a PDZ protein previously identified as interactor of SARS-CoV-1 E protein in epithelia, was also found in our analysis. Little is known about these PDZ proteins in immune cells, so we addressed their expression in monocytes and along the differentiation towards macrophages and dendritic cells. Our findings support the notion that viral targeting of PDZ proteins alters inflammatory response and highlight the importance of identifying the functions of PDZ proteins into maintenance of immune fitness.

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SARS-CoV-2 pathogenesis mediated by E protein's PBM was prevented by modulators of CFTR function

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