Lymphomas driven by Epstein–Barr virus nuclear antigen-1 (EBNA1) are dependant upon Mdm2

Abstract

Epstein–Barr virus (EBV)-associated Burkitt’s lymphoma is characterised by the deregulation of c-Myc expression and a restricted viral gene expression pattern in which the EBV nuclear antigen-1 (EBNA1) is the only viral protein to be consistently expressed. EBNA1 is required for viral genome propagation and segregation during latency. However, it has been much debated whether the protein plays a role in viral-associated tumourigenesis. We show that the lymphomas which arise in EµEBNA1 transgenic mice are unequivocally linked to EBNA1 expression and that both C-Myc and Mdm2 deregulation are central to this process. Tumour cell survival is supported by IL-2 and there is a skew towards CD8-positive T cells in the tumour environment, while the immune check-point protein PD-L1 is upregulated in the tumours. Additionally, several isoforms of Mdm2 are upregulated in the EµEBNA1 tumours, with increased phosphorylation at ser166, an expression pattern not seen in Eµc-Myc transgenic tumours. Concomitantly, E2F1, Xiap, Mta1, C-Fos and Stat1 are upregulated in the tumours. Using four independent inhibitors of Mdm2 we demonstrate that the EµEBNA1 tumour cells are dependant upon Mdm2 for survival (as they are upon c-Myc) and that Mdm2 inhibition is not accompanied by upregulation of p53, instead cell death is linked to loss of E2F1 expression, providing new insight into the underlying tumourigenic mechanism. This opens a new path to combat EBV-associated disease.

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Acknowledgements

Thanks to Daniela Quintana for contribution to cloning the line 26 junctional sequences, Goutham Subramanain for contribution to characterising the Rab28 genome region sequences in the line 26 and Maria Jackson who contributed to construction of the dnEBNA1 plasmids.

Author contributions

SA conducted FACS experiments, western analyses and drug assays, YA-S cloned the line 59 transgene and contributed to FISH, DC and MD contributed to mouse work and cloning the line 26 transgene, AH conducted the dnEBNA1 analyses, SB supervised and analysed the FISH experiment, PH, AK and PL contributed to the RNA data (array and seq) analyses, KA contributed to the drug analyses, MB conducted the PD-L1 analysis, PT contributed to transgene cloning, the array experiment and mouse work. JBW devised, supervised and procured funding for the study and wrote the manuscript.

Funding

Funding contribution to the work includes former LRF (now Bloodwise) grants. AH was supported by a Wellcome Trust PhD studentship while at GU, KA is supported by a PhD BBSRC studentship. SA is, and YA was, supported by Saudi Arabian PhD scholarships.

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Correspondence to Joanna B. Wilson.

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AlQarni, S., Al-Sheikh, Y., Campbell, D. et al. Lymphomas driven by Epstein–Barr virus nuclear antigen-1 (EBNA1) are dependant upon Mdm2. Oncogene 37, 3998–4012 (2018). https://doi.org/10.1038/s41388-018-0147-x

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