Abstract
Epstein–Barr virus (EBV), which is ubiquitous in the adult population, is causally associated with human malignancies. Like many infectious agents, EBV has evolved strategies to block host cell death, including through expression of viral homologues of cellular BCL-2 pro-survival proteins (vBCL-2s), such as BHRF1. Small molecule inhibitors of the cellular pro-survival BCL-2 family proteins, termed ‘BH3-mimetics’, have entered clinical trials for blood cancers with the BCL-2 inhibitor venetoclax already approved for treatment of therapy refractory chronic lymphocytic leukaemia and acute myeloid leukaemia in the elderly. The generation of BH3-mimetics that could specifically target vBCL-2 proteins may be an attractive therapeutic option for virus-associated cancers, since these drugs would be expected to only kill virally infected cells with only minimal side effects on normal healthy tissues. To achieve this, a better understanding of the contribution of vBCL-2 proteins to tumorigenesis and insights into their biochemical functions is needed. In the context of Burkitt lymphoma (BL), BHRF1 expression conferred strong resistance to diverse apoptotic stimuli. Furthermore, BHRF1 expression in mouse haematopoietic stem and progenitor cells accelerated MYC-induced lymphoma development in a model of BL. BHRF1 interacts with the cellular pro-apoptotic BCL-2 proteins, BIM, BID, PUMA and BAK, but its capability to inhibit apoptosis could not be mapped solely to one of these interactions, suggesting plasticity is a key feature of BHRF1. Site-directed mutagenesis revealed a site in BHRF1 that was critical for its interaction with PUMA and blocking DNA-damage-induced apoptosis, identifying a potentially therapeutically targetable vulnerability in BHRF1.
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Acknowledgements
We thank G. Siciliano, H. Johnson, C. Alessandro, K. Hughes and D. Fayle for mouse husbandry, Dr S. Iyer for advice to establish experimental techniques and Prof. T. Speed for advice with statistical testing. This work was supported by funding from the Kay Kendall Leukaemia Fund Fellowship KKL331 awarded to GLK; Victorian Cancer Agency Fellowship (MCRF 17028) awarded to GLK, Cancer Research UK Programme Grant C5575/A15032 awarded to MR; Cancer Council Victoria, grants-in-aid #1086157 and #1147328 awarded to GLK; the National Health and Medical Research Council, Project Grant #1086291 awarded to GLK and #637336 to MK, Program Grant #101671 awarded to PMC, DCSH and AS, Fellowship #1020363 awarded to AS, Fellowship #1116934 awarded to PMC, Fellowship #1043149 awarded to DCSH, Fellowship #637372 awarded to MK; the Leukaemia Foundation Australia grant awarded to GLK and AS, the Leukaemia and Lymphoma Society Grant #7001-13, awarded to PMC, DCSH, AS and MJH; the estate of Anthony (Toni) Redstone OAM and The Craig Perkins Cancer Research Foundation; Australian Research Council Future Fellowship awarded to MK (FT130101349) and operational infrastructure grants through the Australian Government NHMRCS IRIISS and the Victorian State Government Operational Infrastructure Support.
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LF, RC, CC, NS, LCAG, CDS, MK and GLK performed the experiments. GD, MJH, PMC, DCSH, AS, MK, MR and GLK provided resources. LF, MJH, RJT, CS, AIB, DCC, GD, MK, AS, PMC, DCSH, MR and GLK contributed to the conception and design of the research. LF, CC and GLK analysed the data and LF, MK, MR, AS and GLK interpreted the data. GLK wrote the manuscript and it was edited and critically revised by LF, AS, MK and MR.
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The Walter and Eliza Hall Institute receives milestone and royalty payments related to venetoclax, and employees (CC, LCAG, GD, MJH, PMC, DCSH, AS and GLK) may be eligible for benefits related to these payments. The laboratory of AS receives research funding from Servier. The other authors declare that they have no conflict of interest.
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Fitzsimmons, L., Cartlidge, R., Chang, C. et al. EBV BCL-2 homologue BHRF1 drives chemoresistance and lymphomagenesis by inhibiting multiple cellular pro-apoptotic proteins. Cell Death Differ 27, 1554–1568 (2020). https://doi.org/10.1038/s41418-019-0435-1
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DOI: https://doi.org/10.1038/s41418-019-0435-1
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