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Specific inhibition of NLRP3 in chikungunya disease reveals a role for inflammasomes in alphavirus-induced inflammation

Abstract

Mosquito-borne viruses can cause severe inflammatory diseases and there are limited therapeutic solutions targeted specifically at virus-induced inflammation. Chikungunya virus (CHIKV), a re-emerging alphavirus responsible for several outbreaks worldwide in the past decade, causes debilitating joint inflammation and severe pain. Here, we show that CHIKV infection activates the NLRP3 inflammasome in humans and mice. Peripheral blood mononuclear cells isolated from CHIKV-infected patients showed elevated NLRP3, caspase-1 and interleukin-18 messenger RNA expression and, using a mouse model of CHIKV infection, we found that high NLRP3 expression was associated with peak inflammatory symptoms. Inhibition of NLRP3 activation using the small-molecule inhibitor MCC950 resulted in reduced CHIKV-induced inflammation and abrogated osteoclastogenic bone loss and myositis, but did not affect in vivo viral replication. Mice treated with MCC950 displayed lower expression levels of the cytokines interleukin-6, chemokine ligand 2 and tumour necrosis factor in joint tissue. Interestingly, MCC950 treatment abrogated disease signs in mice infected with a related arthritogenic alphavirus, Ross River virus, but not in mice infected with West Nile virus—a flavivirus. Here, using mouse models of alphavirus-induced musculoskeletal disease, we demonstrate that NLRP3 inhibition in vivo can reduce inflammatory pathology and that further development of therapeutic solutions targeting inflammasome function could help treat arboviral diseases.

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Acknowledgements

We thank S. Masters (the Walter and Eliza Hall Institute of Medical Research) for kindly providing the caspase-1–/– and ASC–/– mice. This study was supported by grants from the Australian National Health and Medical Research Council (NHMRC) to S.M. (APP1079086). S.M. is the recipient of an NHMRC Senior Research Fellowship (APP1059167) and A.S. is the recipient of an NHMRC Principal Research Fellowship (APP1058391).

Author information

W.C., L.F.P.N. and S.M. conceptualized and designed the study. W.C., S-S.F., A.Z., S.W., L.J.H., K.T., A.T. and J.R.F. performed the mouse CHIKV and RRV experiments and analysed the data. L.D.V. and C.v.V. performed the mouse WNV experiments and analysed the data. R.W.L. performed the bone computed tomography experiments and analysed the data. T.M.W. and R.G. performed MCC950 pharmacokinetics experiments and analysed the data. D.M.O., H.I.N., T-D.K., L.A.J.O., A.A.B.R., N.J.K., A.S. and M.A.C. provided the materials and analysis methods. A.Z. wrote the manuscript. A.Z., R.W.L., N.J.K., A.S., L.F.P.N., M.A.C. and S.M. reviewed and revised the manuscript.

Competing interests

The authors declare no competing financial interests.

Correspondence to Suresh Mahalingam.

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  1. Supplementary Information

    Supplementary Figures 1–7, Supplementary Table 1, Supplementary References.

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Further reading

Fig. 1: NLRP3, IL-18 and IL-1β expression is highly elevated in CHIKV-infected patients.
Fig. 2: CHIKV infection leads to rapid induction of inflammasomes in the ankle joints of mice.
Fig. 3: Treatment with NLRP3 and caspase-1 inhibitor ameliorates CHIKV-induced footpad swelling.
Fig. 4: Inhibition of inflammasome activation with MCC950 and Z-YVAD-FMK reduces pathological bone loss in CHIKV-infected mice.
Fig. 5: MCC950 and Z-YVAD-FMK treatment suppresses CHIKV-induced osteoclastogenesis.
Fig. 6: MCC950 and Z-YVAD-FMK treatment downregulates the expression of inflammasome components in CHIKV-infected mice.