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APOBEC3A intratumoral DNA electroporation in mice

Abstract

Human APOBEC3A (A3A) cytidine deaminase shows pro-apoptotic properties resulting from hypermutation of genomic DNA, induction of double-stranded DNA breaks (DSBs) and G1 cell cycle arrest. Given this, we evaluated the antitumor efficacy of A3A by intratumoral electroporation of an A3A expression plasmid. DNA was repeatedly electroporated into B16OVA, B16Luc tumors of C57BL/6J mice as well as the aggressive fibrosarcoma Sarc2 tumor of HLA-A*0201/DRB1*0101 transgenic mice using noninvasive plate electrodes. Intratumoral electroporation of A3A plasmid DNA resulted in regression of ~50% of small B16OVA melanoma tumors that did not rebound in the following 2 months without treatment. Larger or more aggressive tumors escaped regression when so treated. As APOBEC3A was much less efficient in provoking hypermutation and DSBs in B16OVA cells compared with human or quail cells, it is likely that APOBEC3A would be more efficient in a human setting than in a mouse model.

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References

  1. Vanneman M, Dranoff G . Combining immunotherapy and targeted therapies in cancer treatment. Nat Rev Cancer 2012; 12: 237–251.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Senkus E, Jassem J . Cardiovascular effects of systemic cancer treatment. Cancer Treat Rev 2011; 37: 300–311.

    Article  CAS  PubMed  Google Scholar 

  3. Niu G, Heller R, Catlett-Falcone R, Coppola D, Jaroszeski M, Dalton W et al. Gene therapy with dominant-negative Stat3 suppresses growth of the murine melanoma B16 tumor in vivo. Cancer Res 1999; 59: 5059–5063.

    CAS  PubMed  Google Scholar 

  4. Goto T, Nishi T, Tamura T, Dev SB, Takeshima H, Kochi M et al. Highly efficient electro-gene therapy of solid tumor by using an expression plasmid for the herpes simplex virus thymidine kinase gene. Proc Natl Acad Sci USA 2000; 97: 354–359.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Daud A, Algazi A, Ashworth M, Buljan M, Takamura KT, Diep T et al. Intratumoral electroporation of plasmid interleukin-12: efficacy and biomarker analyses from a phase 2 study in melanoma (OMS-I100). J Transl Med 2015; 13: 2068.

    Google Scholar 

  6. Chuang TF, Lee SC, Liao KW, Hsiao YW, Lo CH, Chiang BL et al. Electroporation-mediated IL-12 gene therapy in a transplantable canine cancer model. Int J Cancer 2009; 125: 698–707.

    Article  CAS  PubMed  Google Scholar 

  7. Heller L, Pottinger C, Jaroszeski MJ, Gilbert R, Heller R . In vivo electroporation of plasmids encoding GM-CSF or interleukin-2 into existing B16 melanomas combined with electrochemotherapy induces long-term antitumour immunity. Melanoma Res 2000; 10: 577–583.

    Article  CAS  PubMed  Google Scholar 

  8. Daud AI, DeConti RC, Andrews S, Urbas P, Riker AI, Sondak VK et al. Phase I trial of interleukin-12 plasmid electroporation in patients with metastatic melanoma. J Clin Oncol 2008; 26: 5896–5903.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Heller LC, Coppola D . Electrically mediated delivery of vector plasmid DNA elicits an antitumor effect. Gene Therapy 2002; 9: 1321–1325.

    Article  CAS  PubMed  Google Scholar 

  10. Collins CG, Tangney M, Larkin JO, Casey G, Whelan MC, Cashman J et al. Local gene therapy of solid tumors with GM-CSF and B7-1 eradicates both treated and distal tumors. Cancer Gene Ther 2006; 13: 1061–1071.

    Article  CAS  PubMed  Google Scholar 

  11. Bogerd HP, Wiegand HL, Hulme AE, Garcia-Perez JL, O'Shea KS, Moran JV et al. Cellular inhibitors of long interspersed element 1 and Alu retrotransposition. Proc Natl Acad Sci USA 2006; 103: 8780–8785.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Vartanian JP, Guetard D, Henry M, Wain-Hobson S . Evidence for editing of human papillomavirus DNA by APOBEC3 in benign and precancerous lesions. Science 2008; 320: 230–233.

    Article  CAS  PubMed  Google Scholar 

  13. Chen H, Lilley CE, Yu Q, Lee DV, Chou J, Narvaiza I et al. APOBEC3A is a potent inhibitor of adeno-associated virus and retrotransposons. Curr Biol 2006; 16: 480–485.

    Article  CAS  PubMed  Google Scholar 

  14. Stenglein MD, Burns MB, Li M, Lengyel J, Harris RS . APOBEC3 proteins mediate the clearance of foreign DNA from human cells. Nat Struct Mol Biol 2010; 17: 222–229.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Suspene R, Aynaud MM, Guetard D, Henry M, Eckhoff G, Marchio A et al. Somatic hypermutation of human mitochondrial and nuclear DNA by APOBEC3 cytidine deaminases, a pathway for DNA catabolism. Proc Natl Acad Sci USA 2011; 108: 4858–4863.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Caval V, Suspene R, Shapira M, Vartanian JP, Wain-Hobson S . A prevalent cancer susceptibility APOBEC3A hybrid allele bearing APOBEC3B 3'UTR enhances chromosomal DNA damage. Nat Commun 2014; 5: 5129.

    Article  CAS  PubMed  Google Scholar 

  17. Suspene R, Aynaud MM, Vartanian JP, Wain-Hobson S . Efficient deamination of 5-methylcytidine and 5-substituted cytidine residues in DNA by human APOBEC3A cytidine deaminase. PLoS One 2013; 8: e63461.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Koning FA, Newman EN, Kim EY, Kunstman KJ, Wolinsky SM, Malim MH . Defining APOBEC3 expression patterns in human tissues and hematopoietic cell subsets. J Virol 2009; 83: 9474–9485.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Refsland EW, Stenglein MD, Shindo K, Albin JS, Brown WL, Harris RS . Quantitative profiling of the full APOBEC3 mRNA repertoire in lymphocytes and tissues: implications for HIV-1 restriction. Nucleic Acids Res 2010; 38: 4274–4284.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Berger G, Durand S, Fargier G, Nguyen XN, Cordeil S, Bouaziz S et al. APOBEC3A is a specific inhibitor of the early phases of HIV-1 infection in myeloid cells. PLoS Pathog 2011; 7: e1002221.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Caval V, Suspene R, Vartanian JP, Wain-Hobson S . Orthologous mammalian APOBEC3A cytidine deaminases hypermutate nuclear DNA. Mol Biol Evol 2014; 31: 330–340.

    Article  CAS  PubMed  Google Scholar 

  22. Kostrzak A, Henry M, Demoyen PL, Wain-Hobson S, Vartanian JP . APOBEC3A catabolism of electroporated plasmid DNA in mouse muscle. Gene Therapy 2015; 22: 96–103.

    Article  CAS  PubMed  Google Scholar 

  23. Lucas ML, Heller L, Coppola D, Heller R . IL-12 plasmid delivery by in vivo electroporation for the successful treatment of established subcutaneous B16.F10 melanoma. Mol Ther 2002; 5: 668–675.

    Article  CAS  PubMed  Google Scholar 

  24. Li S, Zhang X, Xia X . Regression of tumor growth and induction of long-term antitumor memory by interleukin 12 electro-gene therapy. J Natl Cancer Inst 2002; 94: 762–768.

    Article  CAS  PubMed  Google Scholar 

  25. Steighner RJ, Povirk LF, Bleomycin-induced DNA . lesions at mutational hot spots: implications for the mechanism of double-strand cleavage. Proc Natl Acad Sci USA 1990; 87: 8350–8354.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Kroemer G, Galluzzi L, Kepp O, Zitvogel L . Immunogenic cell death in cancer therapy. Annu Rev Immunol 2013; 31: 51–72.

    Article  CAS  PubMed  Google Scholar 

  27. Krysko DV, Garg AD, Kaczmarek A, Krysko O, Agostinis P, Vandenabeele P . Immunogenic cell death and DAMPs in cancer therapy. Nat Rev Cancer 2012; 12: 860–875.

    Article  CAS  PubMed  Google Scholar 

  28. Calvet CY, Famin D, Andre FM, Mir LM . Electrochemotherapy with bleomycin induces hallmarks of immunogenic cell death in murine colon cancer cells. Oncoimmunology 2014; 3: e28131.

    Article  PubMed  PubMed Central  Google Scholar 

  29. Alexandrov LB, Nik-Zainal S, Wedge DC, Aparicio SA, Behjati S, Biankin AV et al. Signatures of mutational processes in human cancer. Nature 2013; 500: 415–421.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Nik-Zainal S, Wedge DC, Alexandrov LB, Petljak M, Butler AP, Bolli N et al. Association of a germline copy number polymorphism of APOBEC3A and APOBEC3B with burden of putative APOBEC-dependent mutations in breast cancer. Nat Genet 2014; 46: 487–491.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Caval V, Bouzidi MS, Suspene R, Laude H, Dumargne MC, Bashamboo A et al. Molecular basis of the attenuated phenotype of human APOBEC3B DNA mutator enzyme. Nucleic Acids Res 2015; 43: 9340–9349.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Kishida T, Asada H, Satoh E, Tanaka S, Shinya M, Hirai H et al. In vivo electroporation-mediated transfer of interleukin-12 and interleukin-18 genes induces significant antitumor effects against melanoma in mice. Gene Therapy 2001; 8: 1234–1240.

    Article  CAS  PubMed  Google Scholar 

  33. Lohr F, Lo DY, Zaharoff DA, Hu K, Zhang X, Li Y et al. Effective tumor therapy with plasmid-encoded cytokines combined with in vivo electroporation. Cancer Res 2001; 61: 3281–3284.

    CAS  PubMed  Google Scholar 

  34. Radkevich-Brown O, Piechocki MP, Back JB, Weise AM, Pilon-Thomas S, Wei WZ . Intratumoral DNA electroporation induces anti-tumor immunity and tumor regression. Cancer Immunol Immunother 2010; 59: 409–417.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Kishida T, Asada H, Itokawa Y, Yasutomi K, Shin-Ya M, Gojo S et al. Electrochemo-gene therapy of cancer: intratumoral delivery of interleukin-12 gene and bleomycin synergistically induced therapeutic immunity and suppressed subcutaneous and metastatic melanomas in mice. Mol Ther 2003; 8: 738–745.

    Article  CAS  PubMed  Google Scholar 

  36. Golberg A, Rubinsky B . A statistical model for multidimensional irreversible electroporation cell death in tissue. Biomed Eng Online 2010; 9: 13.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Gronevik E, Mathiesen I, Lomo T . Early events of electroporation-mediated intramuscular DNA vaccination potentiate Th1-directed immune responses. J Gene Med 2005; 7: 1246–1254.

    Article  PubMed  Google Scholar 

  38. Chiarella P, Massi E, De Robertis M, Sibilio A, Parrella P, Fazio VM et al. Electroporation of skeletal muscle induces danger signal release and antigen-presenting cell recruitment independently of DNA vaccine administration. Exp Opin Biol Ther 2008; 8: 1645–1657.

    Article  CAS  Google Scholar 

  39. Aurisicchio L, Mancini R, Ciliberto G . Cancer vaccination by electro-gene-transfer. Expert Rev Vaccines 2013; 12: 1127–1137.

    Article  CAS  PubMed  Google Scholar 

  40. Drabick JJ, Glasspool-Malone J, King A, Malone RW . Cutaneous transfection and immune responses to intradermal nucleic acid vaccination are significantly enhanced by in vivo electropermeabilization. Mol Ther 2001; 3: 249–255.

    Article  CAS  PubMed  Google Scholar 

  41. Miklavcic D, Semrov D, Mekid H, Mir LM . A validated model of in vivo electric field distribution in tissues for electrochemotherapy and for DNA electrotransfer for gene therapy. Biochim Biophys Acta 2000; 1523: 73–83.

    Article  CAS  PubMed  Google Scholar 

  42. Mann CJ, Anguela XM, Montane J, Obach M, Roca C, Ruzo A et al. Molecular signature of the immune and tissue response to non-coding plasmid DNA in skeletal muscle after electrotransfer. Gene Therapy 2012; 19: 1177–1186.

    Article  CAS  PubMed  Google Scholar 

  43. Frandsen SK, Gissel H, Hojman P, Tramm T, Eriksen J, Gehl J . Direct therapeutic applications of calcium electroporation to effectively induce tumor necrosis. Cancer Res 2012; 72: 1336–1341.

    Article  CAS  PubMed  Google Scholar 

  44. Abraham RT . Cell cycle checkpoint signaling through the ATM and ATR kinases. Genes Dev 2001; 15: 2177–2196.

    Article  CAS  PubMed  Google Scholar 

  45. Gasser S, Orsulic S, Brown EJ, Raulet DH . The DNA damage pathway regulates innate immune system ligands of the NKG2D receptor. Nature 2005; 436: 1186–1190.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Gourzi P, Leonova T, Papavasiliou FN . A role for activation-induced cytidine deaminase in the host response against a transforming retrovirus. Immunity 2006; 24: 779–786.

    Article  CAS  PubMed  Google Scholar 

  47. Pajot A, Michel ML, Fazilleau N, Pancre V, Auriault C, Ojcius DM et al. A mouse model of human adaptive immune functions: HLA-A2.1-/HLA-DR1-transgenic H-2 class I-/class II-knockout mice. Eur J Immunol 2004; 34: 3060–3069.

    Article  CAS  PubMed  Google Scholar 

  48. Mir LM, Moller PH, Andre F, Gehl J . Electric pulse-mediated gene delivery to various animal tissues. Adv Genet 2005; 54: 83–114.

    Article  CAS  PubMed  Google Scholar 

  49. Gehl J . Electroporation for drug and gene delivery in the clinic: doctors go electric. Methods Mol Biol 2008; 423: 351–359.

    Article  CAS  PubMed  Google Scholar 

  50. Tomayko MM, Reynolds CP . Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother Pharmacol 1989; 24: 148–154.

    Article  CAS  PubMed  Google Scholar 

  51. Wakabayashi A, Nakagawa Y, Shimizu M, Moriya K, Nishiyama Y, Takahashi H . Suppression of an already established tumor growing through activated mucosal CTLs induced by oral administration of tumor antigen with cholera toxin. J Immunol 2008; 180: 4000–4010.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by Invectys.

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Correspondence to A Kostrzak.

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AK, ME, EP, JT, TB, MJ and TH are employees of Invectys. PLD and SWH are co-founders of Invectys, a biotech devoted to therapeutic cancer vaccination. The remaining authors declare no conflict of interest.

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Kostrzak, A., Caval, V., Escande, M. et al. APOBEC3A intratumoral DNA electroporation in mice. Gene Ther 24, 74–83 (2017). https://doi.org/10.1038/gt.2016.77

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