Numb-associated kinases are required for SARS-CoV-2 infection and are cellular targets for therapy

Karim, M.; Saul, S.; Ghita, L.; Sahoo, M. K.; Ye, C.; Bhalla, N.; Jin, J.; Park, J.-G.; Martinez-Gualda, B.; Patrick East, M.; L. Johnson, G.; Pinsky, B. A.; Martinez-Sobrido, L.; Asquith, C. R. M.; Narayanan, A.; De Jonghe, S.; Einav, S.

doi:10.1101/2022.03.18.484178

bioRxiv · 10.1101/2022.03.18.484178

Numb-associated kinases are required for SARS-CoV-2 infection and are cellular targets for therapy

Karim, M.·Saul, S.·Ghita, L.·Sahoo, M. K.·Ye, C.·Bhalla, N.·Jin, J.·Park, J.-G.·Martinez-Gualda, B.·Patrick East, M.·L. Johnson, G.·Pinsky, B. A.·Martinez-Sobrido, L.·Asquith, C. R. M.·Narayanan, A.·De Jonghe, S.·Einav, S.

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Abstract

The coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to pose serious threats to global health. We previously reported that AAK1, BIKE and GAK, members of the Numb-associated kinase family, control intracellular trafficking of multiple RNA viruses during viral entry and assembly/egress. Here, using both genetic and pharmacological approaches, we probe the functional relevance of NAKs for SARS-CoV-2 infection. siRNA-mediated depletion of AAK1, BIKE, GAK, and STK16, the fourth member of the NAK family, suppressed SARS-CoV-2 infection in human lung epithelial cells. Both known and novel small molecules with potent AAK1/BIKE, GAK or STK16 activity suppressed SARS-CoV-2 infection. Moreover, combination treatment with the approved anti-cancer drugs, sunitinib and erlotinib, with potent anti-AAK1/BIKE and GAK activity, respectively, demonstrated synergistic effect against SARS-CoV-2 infection in vitro. Time-of-addition experiments revealed that pharmacological inhibition of AAK1 and BIKE suppressed viral entry as well as late stages of the SARS-CoV-2 life cycle. Lastly, suppression of NAKs expression by siRNAs inhibited entry of both wild type and SARS-CoV-2 pseudovirus. These findings provide insight into the roles of NAKs in SARS-CoV-2 infection and establish a proof-of-principle that pharmacological inhibition of NAKs can be potentially used as a host-targeted approach to treat SARS-CoV-2 with potential implications to other coronaviruses.

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Karim, M., Saul, S., Ghita, L., Sahoo, M. K., Ye, C., Bhalla, N., Jin, J., Park, J.-G., Martinez-Gualda, B., Patrick East, M., L. Johnson, G., Pinsky, B. A., Martinez-Sobrido, L., Asquith, C. R. M., Narayanan, A., De Jonghe, S., Einav, S.. 2022-03-20. Numb-associated kinases are required for SARS-CoV-2 infection and are cellular targets for therapy. https://doi.org/10.1101/2022.03.18.484178

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