Broad and potent antiviral activity of the NAE inhibitor MLN4924.

Vu Thuy Khanh Le-Trilling,Christine D Landsberg,Barbara Sitek,Ingo Drexler,Meike U Rückborn,Adalbert Krawczyk,Dominik A Megger,Wibke Bayer,Sha Tao,Mirko Trilling,Matthias Tenbusch,Benjamin Katschinski

doi:10.1038/srep19977

Abstract

In terms of infected human individuals, herpesviruses range among the most successful virus families. Subclinical herpesviral infections in healthy individuals contrast with life-threatening syndromes under immunocompromising and immunoimmature conditions. Based on our finding that cytomegaloviruses interact with Cullin Roc ubiquitin ligases (CRLs) in the context of interferon antagonism, we systematically assessed viral dependency on CRLs by utilizing the drug MLN4924. CRL activity is regulated through the conjugation of Cullins with the ubiquitin-like molecule Nedd8. By inhibiting the Nedd8-activating Enzyme (NAE), MLN4924 interferes with Nedd8 conjugation and CRL activity. MLN4924 exhibited pronounced antiviral activity against mouse and human cytomegalovirus, herpes simplex virus (HSV)- 1 (including multi-drug resistant clinical isolates), HSV-2, adeno and influenza viruses. Human cytomegalovirus genome amplification was blocked at nanomolar MLN4924 concentrations. Global proteome analyses revealed that MLN4924 blocks cytomegaloviral replication despite increased IE1 amounts. Expression of dominant negative Cullins assigned this IE regulation to defined Cullin molecules and phenocopied the antiviral effect of MLN4924.

Highlights

Observed that common side effects were fatigue and nausea and ≥ 15% of patients reported adverse events but grade 4 adverse events and treatment-related deaths did not occur[13]
NIH3T3 fibroblasts were treated with graded concentrations of MLN4924 and infected with a recombinant MCMV harbouring a luciferase reporter gene under the control of the endogenous MCMV m157 promoter/enhancer
The experiment was repeated on primary murine embryonic fibroblasts (MEF) from C57BL/6 and IFNAR1−/− mice, but no significant difference between the cell types was observed at 2, 4 and 6 days post infection (p > 0.66, p > 0.79 and p > 0.23, respectively - data not shown). These findings indicate that MLN4924 counteracts the activity of the interferon antagonist pM27, it elicits its antiviral activity against MCMV pM27- and type I IFN-independently, suggesting that NAE and CRL activity are required for further layers of protein turnover regulation during viral replication

Summary

Introduction

Observed that common side effects were fatigue and nausea and ≥ 15% of patients reported adverse events but grade 4 adverse events and treatment-related deaths did not occur[13]. We have previously shown that the mouse cytomegalovirus (MCMV)-encoded protein pM27 exploits DNA-damage and DNA-binding (DDB) 1 containing Cullin 4A/B ubiquitin ligase complexes to induces (poly-) ubiquitination and subsequent proteasomal degradation of STAT214. We show that MCMV replication is severely compromised in the presence of MLN4924, displaying viral dependency on NAE and CRLs. Besides the effect on MCMV, we found potent antiviral activity of MLN4924 against the human-pathogenic viruses HCMV, Herpes simplex virus (HSV)-1, HSV-2, adenovirus 5 (AdV5) and influenza virus PR8, but not against vaccinia virus (VACV) and vesicular stomatitis virus (VSV). Our findings uncover NAE and CRLs as essential and druggable host factors for the replication of several clinically relevant viruses

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Conclusion