Abstract
Epstein–Barr nuclear antigen 1 (EBNA1), a dimeric oncoprotein of the Epstein–Barr virus (EBV), is essential for both viral-genome maintenance and the survival of infected cells. Despite EBNA1’s potential as a therapeutic target, tools for the direct monitoring of EBNA1 in vitro and in vivo are lacking. Here, we show that a peptide-based inhibitor that luminesces when bound to EBNA1 inside the nucleus of EBV+ cells can regulate EBNA1 homodimer formation and selectively inhibit the growth of EBV+ tumours of nasopharyngeal carcinoma cells (C666-1 and NPC43) and Burkitt’s lymphoma Raji cells. We also show that the peptide-based probe leads to 93% growth inhibition of EBV+ tumours in mice. Our findings support the hypothesis that selective inhibition of EBNA1 dimerization can be used to afford better EBV-related cancer differentiation, and highlight the potential application of the probe as a new generation of biotracers for investigating the fundamental biological function of EBNA1 and for exploring its application as a therapeutic target.
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References
Cohen, J. I. Epstein–Barr virus infection. N. Eng. J. Med. 343, 481–492 (2000).
Young, L. S. & Rickinson, A. B. Epstein–Barr virus: 40 years on. Nat. Rev. Cancer 4, 757–768 (2004).
Thorley-Lawson, D. A. Epstein–Barr virus: exploiting the immune system. Nat. Rev. Immunol. 1, 75–82 (2001).
Paludan, S. R., Bowie A. G., Horan K. A. & Fitzgerald K. A. Recognition of herpesviruses by the innate immune system. Nat. Rev. Immunol. 11, 143–154 (2011).
Wood, V. H. J. et al. Epstein–Barr virus-encoded EBNA1 regulates cellular gene transcription and modulates the STAT1 and TGFb signalling pathways. Oncogene 26, 4135–4147 (2007).
Frappier, L. The Epstein–Barr virus EBNA1 protein. Scientifica 2012, 438204 (2012).
Li, N., Thompson, S., Jiang, H., Liberman, P. M. & Luo, C. Development of drugs for Epstein–Barr virus using high-throughput in silico virtual screening. Expert Opin. Drug Discov. 5, 1189–1203 (2010).
Kim, S. Y. et al. Small molecule and peptide-mediate inhibition of Epstein–Barr virus nuclear antigen 1 dimerization. Biochem. Biophys. Res. Commun. 424, 251–256 (2012).
Jiang, L. et al. EBNA1-specific luminescent small molecules for the imaging and inhibition of latent EBV-infected tumor cells. Chem. Commun. 50, 6517–6519 (2014).
Fisher, N., Vo, M. D., Mueller-Lantzsch, N. & Grässer, F. A. A potential NES of the Epstein–Barr virus nuclear antigen 1 (EBNA1) does not confer shuttling. FEBS Lett. 447, 311–314 (1999).
Kanda, T., Otter, M. & Wahl, G. M. Coupling of mitotic chromosome tethering and replication competence in Epstein–Barr virus-based plasmids. Mol. Cell. Biol. 21, 3576–3588 (2001).
Vacik, J., Dean, B. S., Zimmer, W. E. & Dean, D. A. Cell-specific nuclear import of plasmid DNA. Gene Ther. 6, 1006–1014 (1999).
Bochkarev, A., Bochkareva, E., Frappier, L. & Edwards, A. M. The 2.2 Å structure of a permanganate-sensitive DNA site bound by the Epstein–Barr virus origin binding protein, EBNA1. J. Mol. Biol. 284, 1273–1278 (1998).
Correia, B. et al. Crystal structure of the gamma-2 herpesvirus LANA DNA binding domain identifies charged surface residues which impact viral latency. PLoS Pathog. 9, e1003673 (2013).
Case, D. A. et al. Amber 14 (Univ. California, San Francisco, 2014).
Lippert, E. et al. Umwandlung von elektronenanregungsenergie. Angew. Chem. 73, 695–706 (1961).
Grabowski, Z. R. & Rotkiewicz, K. Structural changes accompanying intramolecular electron transfer: focus on twisted intramolecular charge-transfer states and structures. Chem. Rev. 103, 3899–4031(2003).
Sahoo, D. & Chakravorti, S. Dye-surfactant interaction: modulation of photophysics of an ionic styryl dye. Photochem. Photobiol. 85, 1103–1109 (2009).
Wandelt, B., Mielniczak, A., Turkewitsch, P., Darling, G. D. & Stranix, B. R. Substituted 4-[4-(dimethylamino)styryl] pyridinium salt as a fluorescent probe for cell microviscosity. Biosens. Bioelectron. 18, 465–471 (2003).
Chakraborty, A., Kar, S. & Guchhait, N. Secondary amino group as charge donor for the excited state intramolecular charge transfer reaction in trans-3-(4-monomethylamino-phenyl)-acrylic acid: spectroscopic measurement and theoretical calculations. J. Photochem. Photobiol. A 181, 246–256 (2006).
Mahon, K. P. et al. Deconvolution of the cellular oxidative stress response with organelle-specific peptide conjugates. Chem. Biol. 14, 923–930 (2007).
Puckett, C. A. & Barton, J. K. Targeting a ruthenium complex to the nucleus with short peptides. Bioorg. Med. Chem. 18, 3564–3569 (2010).
Lindorff-Larsen, K. et al. Improved side-chain torsion potentials for the amber ff99SB protein. Proteins 78, 1950–1958 (2010).
Jorgensen, W. L., Chandrasekhar, J., Madura, J. D., Impey, R. W. & Klein, M. L. Comparison of simple potential functions for simulating liquid water. J. Chem. Phys. 79, 926–935 (1983).
Acknowledgements
This work is funded by the Hong Kong Baptist University (FRG2/14-15/013013), Hong Kong Polytechnic University (HKPolyU), Hong Kong Research Grants Council (HKBU 20301615), Hong Kong Polytechnic University Central Research Grant (G-UC08), Hong Kong Research Grants Council (PolyU 153012/15P), University Research Facility for Chemical and Environmental Analysis (UCEA) and Area of Excellent Grants (1-ZVGG) of Hong Kong Polytechnic University, ECS-Grant - RGC (PolyU 253002/14P), HK PolyU (PolyU 5096/13P), HKBU and HKPolyU Joint Research Programme (RC-ICRS/15-16/02F-WKL02F-WKL), the EPSRC (Durham University, DTA award) and Research Grants Council of the Hong Kong SAR for the NPC Area of Excellence (AoE/M 06/08 Center for Nasopharyngeal Carcinoma Research).
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N.-K.M. and K.-L.W. conceived and supervised the project. L.J. performed the synthesis, characterization, spectroscopic properties measurements and cytotoxicity assays in normal and some EBV-negative cells; H.L. contributed the photophysical data analysis. R.L. purified the WT-EBNA1 and EBNA1 mutants, and conducted the dimerization assay; T.H., Z.-X.B. and W.-T.W. contributed to the MD simulations; C.-F.C. carried out the confocal imaging, co-localization and some cytotoxicity assays; S.L., J.Z. and S.L.C. performed the peptide synthesis and purified the peptide conjugates; W.-Y.W., M.M.-L.L., B.D.C. and W.C.S.T. carried out the in vivo inhibition. M.L.L., H.L.L., S.W.T. and G.S.T. contributed to the in vitro cytotoxicity in EBV-positive and EBV-negative pairs of cancer cell lines; G.-L.L. participated in the cell work and molecular design; L.J., W.-L.C., W.-S.L., G.-L.L., S.L.C. and K.-L.W. wrote the manuscript.
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Supplementary methods, figures, tables and references. (PDF 11131 kb)
Supplementary Video 1
Dynamic distribution of L2P4 in EBV-negative HeLa cells. (MOV 2419 kb)
Supplementary Video 2
Dynamic distribution of L2P4 in EBV-positive C666-1 cells. (MOV 2723 kb)
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Jiang, L., Lan, R., Huang, T. et al. EBNA1-targeted probe for the imaging and growth inhibition of tumours associated with the Epstein–Barr virus. Nat Biomed Eng 1, 0042 (2017). https://doi.org/10.1038/s41551-017-0042
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DOI: https://doi.org/10.1038/s41551-017-0042
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