Norovirus is a major cause of acute gastroenteritis worldwide and has emerged as an important issue of chronic infection in transplantation patients. Since no approved antiviral is available, we evaluated the effects of different immunosuppressants and ribavirin on norovirus and explored their mechanisms of action by using a human norovirus (HuNV) replicon-harboring model and a surrogate murine norovirus (MNV) infectious model. The roles of the corresponding drug targets were investigated by gain- or loss-of-function approaches. We found that the calcineurin inhibitors cyclosporine (CsA) and tacrolimus (FK506) moderately inhibited HuNV replication. Gene silencing of their cellular targets, cyclophilin A, FKBP12, and calcineurin, significantly inhibited HuNV replication. A low concentration, therapeutically speaking, of mycophenolic acid (MPA), an uncompetitive IMP dehydrogenase (IMPDH) inhibitor, potently and rapidly inhibited norovirus replication and ultimately cleared HuNV replicons without inducible resistance following long-term drug exposure. Knockdown of the MPA cellular targets IMPDH1 and IMPDH2 suppressed HuNV replication. Consistent with the nucleotide-synthesizing function of IMPDH, exogenous guanosine counteracted the antinorovirus effects of MPA. Furthermore, the competitive IMPDH inhibitor ribavirin efficiently inhibited norovirus and resulted in an additive effect when combined with immunosuppressants. The results from this study demonstrate that calcineurin phosphatase activity and IMPDH guanine synthase activity are crucial in sustaining norovirus infection; thus, they can be therapeutically targeted. Our results suggest that MPA shall be preferentially considered immunosuppressive medication for transplantation patients at risk of norovirus infection, whereas ribavirin represents as a potential antiviral for both immunocompromised and immunocompetent patients with norovirus gastroenteritis.
Bibliographical noteFunding Information:
We declare no conflicts of interest. This work was supported by the Dutch Digestive Foundation (MLDS) for a career development grant (no. CDG 1304) and the China Scholarship Council for funding Ph.D. fellowships to W. Dang (grant 201406180072) and Y. Yin (grant 201307720045).
We gratefully acknowledge Herbert W. Virgin (Washington University, St. Louis, MO, USA) for providing us the MNV-1 and Kyeong-Ok Chang (Kansas State University, USA) for providing the HuNV replicon. We also thank the general support of the Center for Drug Design (University of Minnesota, USA) for developing the IMPDH inhibitors. We declare no conflicts of interest. This work was supported by the Dutch Digestive Foundation (MLDS) for a career development grant (no. CDG 1304) and the China Scholarship Council for funding Ph.D. fellowships to W. Dang (grant 201406180072) and Y. Yin (grant 201307720045).
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- Calcineurin inhibitors
- Cell culture
- Cell culture model
- Mycophenolic acid