Journal: Cell Reports

Article Title: Antiviral drug screen identifies DNA-damage response inhibitor as potent blocker of SARS-CoV-2 replication

doi: 10.1016/j.celrep.2021.108940

Figure Lengend Snippet: Berzosertib inhibits SARS-CoV-2, SARS-CoV-1, and MERS-CoV replication in human cells and is synergistic with remdesivir (A and B) Graphs show an eight-point dose-response curve of berzosertib (A) or remdesivir (B) in SARS-CoV-2-infected Calu-3 cells. Contrasted with cell viability of mock-infected cells. (C and D) Antiviral effect of berzosertib on SARS-CoV-1 (C) and MERS-CoV (D). (E) Graphs show antiviral activity measured with a SARS-CoV-2 immunostaining signal used for identification of infected A549-ACE2 cells. IC 50 values were calculated by non-linear regression sigmoidal dose-response analysis using the GraphPad Prism 7 software package. (F) Graph shows synergistic effect of berzosertib and remdesivir in infected A549-ACE2 cells. Dose-response curves obtained with mixtures of remdesivir and berzosertib, remdesivir alone (thick black line), and berzosertib alone (thick pink line) are shown. (G) Isobologram of drug combinations is depicted. (H) Combinatorial data were analyzed for inhibitory, additive, or synergistic effects (upper triangle, dotted line, and lower triangle, respectively) by using the Compusyn software package.

Article Snippet: IC 50 values were calculated by non-linear regression sigmoidal dose-response analysis using the GraphPad Prism 7 software package. (F) Graph shows synergistic effect of berzosertib and remdesivir in infected A549-ACE2 cells.

Techniques: Infection, Activity Assay, Immunostaining, Software

Journal: Cell Reports

Article Title: Antiviral drug screen identifies DNA-damage response inhibitor as potent blocker of SARS-CoV-2 replication

doi: 10.1016/j.celrep.2021.108940

Figure Lengend Snippet: Berzosertib inhibits SARS-CoV-2 replication in hiPSC-CMs (A) Graph shows beats per minute of SARS-CoV-2-infected hiPSC-CM cells treated with berzosertib (250 nM), dactolisib (250 nM), remdesivir (10 μM), and HQ (10 μM). (B) Graph shows viral titer (TCID 50 /mL) of supernatant collected at the indicated time points after SARS-CoV-2 infection of drug-treated hiPSC-CMs. (C) Graph depicts quantification of SARS-CoV-2 and cleaved caspase-3-positive cells. (D) IFA images of hiPSC-CMs undergoing apoptosis after SARS-CoV-2 infection and drug treatment at 72 hpi. Scale bar, 25 μm. (E) hiPSC-CMs were stained with cardiac troponin T (cTnT) (green) to demonstrate that cells are protected from SARS-CoV-2-mediated cell injury (red) by berzosertib (250 nM). Scale bar, 25 μm. Statistical analysis of graphs (A and C) was conducted by multiple-comparison one-way analysis of variance (ANOVA) was conducted. ∗∗ p < 0.001, ∗∗∗ p < 0.0001. Representative data from three independent experiments are presented.

Article Snippet: IC 50 values were calculated by non-linear regression sigmoidal dose-response analysis using the GraphPad Prism 7 software package. (F) Graph shows synergistic effect of berzosertib and remdesivir in infected A549-ACE2 cells.

Techniques: Infection, Staining

Journal: Cell Reports

Article Title: Antiviral drug screen identifies DNA-damage response inhibitor as potent blocker of SARS-CoV-2 replication

doi: 10.1016/j.celrep.2021.108940

Figure Lengend Snippet: Berzosertib mode of antiviral activity in lung and kidney epithelial cells and effect on SARS-CoV-2-mediated inflammatory response (A) Graph shows eight-dose-response curve of berzosertib in SARS-CoV-2-infected human primary lung ALI culture. (B) Immunofluorescent images indicate dose-dependent reduction of SARS-CoV-2 replication in berzosertib-treated ALI culture (spike protein in red).Scale bar, 100 μm. (C) Western blot analysis shows time course of pCHK1 and virus replication kinetics in Vero E6 cells. Berzosertib treatment reduced CHK1 phosphorylation. In addition, it inhibited SARS-CoV-2 replication as early as 8 h after infection. By 24 h in untreated cells, SARS-CoV-2 signal intensity was oversaturated because of the high-level of viral proteins. Representative data from two independent experiments is shown. (D) SARS-CoV-2 genome replication kinetics in the presence of berzosertib treatment on Vero E6 cells. (E) Graph shows that berzosertib treatment reduces the expression of inflammatory IL-6 gene in SARS-CoV-2-infected Vero E6 cells. Statistical analysis of graphs (D and E) conducted with multiple-comparison two-way analysis of variance (ANOVA). ∗ p < 0.01, ∗∗ p < 0.001, ∗∗∗ p < 0.0001. Representative data from three independent experiments are presented.

Article Snippet: IC 50 values were calculated by non-linear regression sigmoidal dose-response analysis using the GraphPad Prism 7 software package. (F) Graph shows synergistic effect of berzosertib and remdesivir in infected A549-ACE2 cells.

Techniques: Activity Assay, Infection, Western Blot, Expressing

Journal: Cell Reports

Article Title: Antiviral drug screen identifies DNA-damage response inhibitor as potent blocker of SARS-CoV-2 replication

doi: 10.1016/j.celrep.2021.108940

Figure Lengend Snippet:

Article Snippet: IC 50 values were calculated by non-linear regression sigmoidal dose-response analysis using the GraphPad Prism 7 software package. (F) Graph shows synergistic effect of berzosertib and remdesivir in infected A549-ACE2 cells.

Techniques: Produced, Recombinant, Electron Microscopy, Blocking Assay, MTT Cell Proliferation, Proliferation Assay, Software