Anticancer pan-ErbB inhibitors reduce inflammation and tissue injury and exert broad-spectrum antiviral effects
Sirle Saul, Marwah Karim, Luca Ghita, Pei-Tzu Huang, Winston Chiu, Verónica Durán, Chieh-Wen Lo, Sathish Kumar, Nishank Bhalla, Pieter Leyssen, Farhang Alem, Niloufar A. Boghdeh, Do H.N. Tran, Courtney A. Cohen, Jacquelyn A. Brown, Kathleen E. Huie, Courtney Tindle, Mamdouh Sibai, Chengjin Ye, Ahmed Magdy Khalil, Kevin Chiem, Luis Martinez-Sobrido, John M. Dye, Benjamin A. Pinsky, Pradipta Ghosh, Soumita Das, David E. Solow-Cordero, Jing Jin, John P. Wikswo, Dirk Jochmans, Johan Neyts, Steven De Jonghe, Aarthi Narayanan, Shirit Einav
Sirle Saul, Marwah Karim, Luca Ghita, Pei-Tzu Huang, Winston Chiu, Verónica Durán, Chieh-Wen Lo, Sathish Kumar, Nishank Bhalla, Pieter Leyssen, Farhang Alem, Niloufar A. Boghdeh, Do H.N. Tran, Courtney A. Cohen, Jacquelyn A. Brown, Kathleen E. Huie, Courtney Tindle, Mamdouh Sibai, Chengjin Ye, Ahmed Magdy Khalil, Kevin Chiem, Luis Martinez-Sobrido, John M. Dye, Benjamin A. Pinsky, Pradipta Ghosh, Soumita Das, David E. Solow-Cordero, Jing Jin, John P. Wikswo, Dirk Jochmans, Johan Neyts, Steven De Jonghe, Aarthi Narayanan, Shirit Einav
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Research Article Virology

Anticancer pan-ErbB inhibitors reduce inflammation and tissue injury and exert broad-spectrum antiviral effects

Abstract

Targeting host factors exploited by multiple viruses could offer broad-spectrum solutions for pandemic preparedness. Seventeen candidates targeting diverse functions emerged in a screen of 4,413 compounds for SARS-CoV-2 inhibitors. We demonstrated that lapatinib and other approved inhibitors of the ErbB family of receptor tyrosine kinases suppress replication of SARS-CoV-2, Venezuelan equine encephalitis virus (VEEV), and other emerging viruses with a high barrier to resistance. Lapatinib suppressed SARS-CoV-2 entry and later stages of the viral life cycle and showed synergistic effect with the direct-acting antiviral nirmatrelvir. We discovered that ErbB1, ErbB2, and ErbB4 bind SARS-CoV-2 S1 protein and regulate viral and ACE2 internalization, and they are required for VEEV infection. In human lung organoids, lapatinib protected from SARS-CoV-2–induced activation of ErbB-regulated pathways implicated in non-infectious lung injury, proinflammatory cytokine production, and epithelial barrier injury. Lapatinib suppressed VEEV replication, cytokine production, and disruption of blood-brain barrier integrity in microfluidics-based human neurovascular units, and reduced mortality in a lethal infection murine model. We validated lapatinib-mediated inhibition of ErbB activity as an important mechanism of antiviral action. These findings reveal regulation of viral replication, inflammation, and tissue injury via ErbBs and establish a proof of principle for a repurposed, ErbB-targeted approach to combat emerging viruses.

Authors

Sirle Saul, Marwah Karim, Luca Ghita, Pei-Tzu Huang, Winston Chiu, Verónica Durán, Chieh-Wen Lo, Sathish Kumar, Nishank Bhalla, Pieter Leyssen, Farhang Alem, Niloufar A. Boghdeh, Do H.N. Tran, Courtney A. Cohen, Jacquelyn A. Brown, Kathleen E. Huie, Courtney Tindle, Mamdouh Sibai, Chengjin Ye, Ahmed Magdy Khalil, Kevin Chiem, Luis Martinez-Sobrido, John M. Dye, Benjamin A. Pinsky, Pradipta Ghosh, Soumita Das, David E. Solow-Cordero, Jing Jin, John P. Wikswo, Dirk Jochmans, Johan Neyts, Steven De Jonghe, Aarthi Narayanan, Shirit Einav

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Figure 1

High-throughput screening for compounds that counteract SARS-CoV-2–induced lethality and validation by plaque assays.

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High-throughput screening for compounds that counteract SARS-CoV-2–induc...
(A) Schematic of the composition of the screened libraries and screening and hit selection pipeline. LOPAC, Library of Pharmacologically Active Compounds (Sigma-Aldrich). (B) High-throughput screening (HTS) assay schematic. Compounds were pre-spotted in 384-well plates at a final concentration of 10 μM and incubated with Vero E6 cells constitutively expressing eGFP for 20 hours, followed by SARS-CoV-2 infection (Belgium-GHB-03021, MOI = 0.001). eGFP signal measured at 4 days after infection was used as an indicator for survival from virus-induced lethality. (C) Box plots of the percentage of fluorescence area values combining the entire HTS data set (2 independent experiments) split into the 4 indicated categories. The box horizontal lines indicate the first, second (median), and third quartiles. Outliers above a cutoff of 15% were defined as positive hits. Dots represent individual compounds, and colors denote positive controls (purple), new hits (blue), and ErbB inhibitors (peach). (D) Heatmap of the EC50 and CC50 values of hits emerging in the HTS, color-coded based on the antiviral activity measured by plaque assays (green) and toxicity measured by alamarBlue assays (orange), 24 hours after infection of Vero cells with SARS-CoV-2 (USA-WA1/2020 strain, MOI = 0.05). Selectivity indices (SI) greater than 5 are depicted in yellow. “#” indicates compounds from the 13-kinase set. (E) Dose-response curves of representative hits depicting SARS-CoV-2 infection (black) and cell viability (blue). Data are relative to DMSO. Data in E are combined from 2 independent experiments, each with 2–3 biological replicates. Means ± SD are shown. Asterisks in A denote 18 hits screened for SARS-CoV-2, VEEV (TC-83), and DENV2.