Fig. 4: Small-molecule inhibition of the SMARCA4 ATPase of mSWI/SNF complexes downregulates ACE2 expression and blocks SARS-CoV-2 infection.
From: Pharmacological disruption of mSWI/SNF complex activity restricts SARS-CoV-2 infection
a, Top: chemical structures of the mSWI/SNF SMARCA4/2 ATPase inhibitors, Comp12 and Comp14. Bottom: three-dimensional structure highlighting Comp12 docked in the ATPase site of the SMARCA2/4 ATPase subunit (Protein Data Bank ID: 6EG2). b, Vero E6 cells were treated with 1.25 μM Comp12 for the indicated times. ACE2 mRNA and protein levels were measured using RT–qPCR and immunoblot, respectively. c, Vero E6 cells were pretreated with Comp12 inhibitors for 2 d and then infected with SARS-CoV-2 at an MOI of 0.2. Cell viability was measured 3 d after infection. d, Vero E6 cells were pretreated with 1.25 and 2.5 μM Comp12 for 2 d and then infected with SARS-CoV-2 pseudovirus. The ACE2 antibody was preincubated with cells for 1 h before infection as a positive control. Luciferase relative to VSVpp-VSV-G control was measured 1 d after infection. e, SARS-CoV-2 production in Comp12 pretreated Vero E6 cells with the indicated concentrations of Comp12 for 2 d. f, ACE2 transcript and protein levels in Comp12-treated Vero E6, Huh7.5 and Calu-3 cells for 2 d at 1.25 and 2.5 μM. g, ACE2 transcript and protein levels in inhibitor and degrader-treated Vero and Huh7.5 cells for 2 d at 1.25 and 2.5 μM. h, Vero E6 and Huh7.5 cells were pretreated with the indicated inhibitors and/or degraders at 2.5 μM for 2 d and then infected with icSARS-CoV-2-mNG. The frequency of infected cells was measured by mNeonGreen expression. i, Vero E6 cells were pretreated with 2.5 μΜ Comp12 for 2 d and then infected with the indicated SARS-CoV-2 variants at an MOI of 0.2. Cell viability was measured 3 d after infection. j, Vero E6 or Calu-3 cells were pretreated with 2.5 μΜ of Comp12 for 2 d and then infected with the SARS-CoV-2 WA1 and E802D viruses. Virus production was measured by plaque assays 1 d after infection. Data in b–j were analyzed using a one-way ANOVA with Tukey’s multiple comparisons test. The mean ± s.e.m. are shown. *P < 0.05, **P < 0.01, ***P < 0.001, n = 3 biological replicates.
