Rock1 is a novel host dependency factor of human enterovirus A71: Implication as a drug target

Abstract Human enterovirus A71 (EV‐A71) is the major causative agent of hand‐foot‐and‐mouth disease (HFMD) commonly associated with severe neurological diseases, particularly in children under 5 years of age. Several investigational therapeutic agents and vaccine candidates are being developed. However, no approved drug against EV‐A71 infection is available, and no proven drug target has been identified. Since host kinases are key regulators of multiple signaling pathways in response to viral infections, here we screened a kinase inhibitor library and identified potent inhibitors against EV‐A71 infection. Among the hits, GSK269962A, a Rho Associated Coiled‐Coil Containing Protein Kinase (Rock) inhibitor with potent antiviral activity, was selected for further analysis. We found that this Rock inhibitor not only efficiently suppressed the replication of EV‐A71 in RD cells, but also in human intestinal organoids, in a dose‐dependent manner. Interestingly, small interfering RNA depletion of Rock1, but not Rock2, significantly restricted viral replication in RD cells, indicating that Rock1 is a novel host dependency factor for EV‐A71 replication and can serve as a target for the development of anti‐EV‐A71 therapeutics.