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Effective gene transfer to solid tumors using different nonviral gene delivery techniques: Electroporation, liposomes, and integrin-targeted vector

Cancer Gene Therapy volume 9, pages 399–406 (2002)Cite this article

Abstract

In this study, we measured transfection efficiency in vitro and in vivo using the following nonviral approaches of gene delivery: injection of plasmid DNA, electroporation-assisted, liposome-enhanced, and integrin-targeted gene delivery, as well as the combination of these methods. Four histologically different tumor models were transfected with a plasmid encoding the green fluorescent protein (GFP) (B16 mouse melanoma, P22 rat carcinosarcoma, SaF mouse sarcoma, and T24 human bladder carcinoma) using adherent cells, dense cell suspensions, and solid tumors. Emphasis was placed on different electroporation conditions to optimise the duration and amplitude of the electric pulses, as well as on different DNA concentrations for effective gene delivery. In addition, transfection efficiency was correlated with cell density of the tumors. The major in vivo findings were: (a) electroporation-assisted gene delivery with plasmid DNA, employing long electric pulses with low amplitude, yielded significantly better GFP expression than short electric pulses with high amplitude; (b) electroporation combined with liposome–DNA complexes yielded the highest percentage of transfected tumor area in B16F1 tumor (6%); (c) transfection efficiency of electroporation-assisted plasmid DNA delivery was dependent on tumor type; (d) integrin-targeted vector, alone or combined with electroporation, was largely ineffective. In conclusion, our results demonstrate that some nonviral methods of gene delivery are feasible and efficient in transfecting solid tumors. Therefore, this makes nonviral methods attractive for further development.

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References

  1. Harvey BG, Worgall S, Ely S et al. Cellular immune responses of healthy individuals to intradermal administration of an E1–E3–adenovirus gene transfer vector Hum Gene Ther 1999 10: 2823–2837

    Article  CAS  Google Scholar 

  2. Marshall E . Gene therapy death prompts review of adenovirus vector Science 1999 286: 2244–2245

    Article  CAS  Google Scholar 

  3. Cotten M, Wagner E . Non-viral approaches to gene therapy Curr Opin Biotechnol 1993 4: 705–710

    Article  CAS  Google Scholar 

  4. Kikuchi A, Aoki Y, Sugaya S et al. Development of novel cationic liposomes for efficient gene transfer into peritoneal disseminated tumor Hum Gene Ther 1999 10: 947–955

    Article  CAS  Google Scholar 

  5. Curiel DT, Agarwal S, Wagner E et al. Adenovirus enhancement of transferrin–polylysine–mediated gene delivery Proc Natl Acad Sci USA 1991 88: 8850–8854

    Article  CAS  Google Scholar 

  6. Wagner E, Plank C, Zatloukal K et al. Influenza virus hemagglutinin HA-2 N-terminal fusogenic peptides augment gene transfer by transferrin–polylysine–DNA complexes: toward a synthetic virus-like gene-transfer vehicle Proc Natl Acad Sci USA 1992 89: 7934–7938

    Article  CAS  Google Scholar 

  7. Simoes S, Slepushkin V, Gaspar R et al. Gene delivery by negatively charged ternary complexes of DNA, cationic liposomes and transferrin or fusogenic peptides Gene Ther 1998 5: 955–964

    Article  CAS  Google Scholar 

  8. Hart SL, Arancibia-Carcamo CV, Wolfert MA et al. Lipid-mediated enhancement of transfection by a nonviral integrin-targeting vector Hum Gene Ther 1998 9: 575–585

    Article  CAS  Google Scholar 

  9. Dachs GU, Coralli C, Hart SL et al. Gene delivery to hypoxic cells in vitro Br J Cancer 2000 83: 662–667

    Article  CAS  Google Scholar 

  10. Jenkins RG, Herrick SE, Meng QH et al. An integrin-targeted non-viral vector for pulmonary gene therapy Gene Ther 2000 7: 393–400

    Article  CAS  Google Scholar 

  11. Mir LM, Glass LF, Sersa G et al. Effective treatment of cutaneous and subcutaneous malignant tumors by electrochemotherapy Br J Cancer 1998 77: 2336–2342

    Article  CAS  Google Scholar 

  12. Sersa G, Stabuc B, Cemazar M et al. Electrochemotherapy with cisplatin: clinical experience in malignant melanoma patients Clin Cancer Res 2000 6: 863–867

    CAS  PubMed  Google Scholar 

  13. Somiari S, Glasspool-Malone J, Drabick JJ et al. Theory and in vivo application of electroporative gene delivery Mol Ther 2000 2: 178–187

    Article  CAS  Google Scholar 

  14. Mir LM, Bureau MF, Gehl J et al. High-efficiency gene transfer into skeletal muscle mediated by electric pulses Proc Natl Acad Sci USA 1999 96: 4262–4267

    Article  CAS  Google Scholar 

  15. Heller L, Jaroszeski MJ, Coppola D et al. Electrically mediated plasmid DNA delivery to hepatocellular carcinomas in vivo Gene Ther 2000 7: 826–829

    Article  CAS  Google Scholar 

  16. Tozer GM, Shaffi KM . Modification of tumour blood flow using the hypertensive agent, angiotensin II Br J Cancer 1993 67: 981–988

    Article  CAS  Google Scholar 

  17. Hill SA, Collinrigde DR, Vojnovic B et al. Tumour radiosensitization by high-oxygen-content gases: influence of the carbon dioxide, content of the inspired gas on pO2, microcirculatory function and radiosensitivity Int J Radiat Oncol Biol Phys 1998 40: 943–951

    Article  CAS  Google Scholar 

  18. Cemazar M, Milacic R, Miklavcic D et al. Intratumoral cisplatin administration in electrochemotherapy: antitumor effectiveness, sequence dependence and platinum content Anticancer Drugs 1998 9: 525–530

    Article  CAS  Google Scholar 

  19. Miklavcic D, Beravs K, Semrov D et al. The importance of electric field distribution for effective in vivo electroporation of tissues Biophys J 1998 74: 2152–2158

    Article  CAS  Google Scholar 

  20. Cemazar M, Miklavcic D, Vodovnik L et al. Improved therapeutic effect of electrochemotherapy by intratumoral drug administration and changing of electrode orientation for electropermeabilization on EAT tumor model in mice Radiol Oncol 1995 29: 121–127

    Google Scholar 

  21. Nabel GJ, Nabel EG, Jang Z et al. Direct gene transfer with DNA–liposome complexes in melanoma: expression, biologic activity, and lack of toxicity in humans Proc Natl Acad Sci USA 1993 90: 11307–11311

    Article  CAS  Google Scholar 

  22. Larchian WA, Horiguchi J, Nair SK et al. Effectiveness of combines interleukin-2 and B7.1 vaccination strategy is dependent on the sequence and order: a liposome-mediated gene therapy treatment for bladder cancer Clin Cancer Res 2000 6: 2913–2920

    CAS  PubMed  Google Scholar 

  23. Yoo GH, Hung MC, Lopez-Berenstein G et al. Phase I trial of intratumoral liposome EiA gene therapy in patients with recurrent breast and head and neck cancer Clin Cancer Res 2001 7: 1237–1245

    CAS  PubMed  Google Scholar 

  24. Wells JM, Li LH, Sen A et al. Electroporation-enhanced gene delivery in mammary tumors Gene Ther 2000 7: 541–547

    Article  CAS  Google Scholar 

  25. Heller R, Schultz J, Lucas ML et al. Intradermal delivery of interleukin-12 plasmid DNA by in vivo electroporation DNA Cell Biol 2001 20: 21–26

    Article  CAS  Google Scholar 

  26. Heller L, Pottinger C, Jaroszeski MJ et al. 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  Google Scholar 

  27. Niu G, Heller R, Catlett-Falcone R 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 

  28. Yamashita YI, Shimada M, Hasegawa H et al. Electroporation-mediated interleukin-12 gene therapy for hepatocellular carcinoma in the mice model Cancer Res 2001 61: 1005–1012

    CAS  PubMed  Google Scholar 

  29. Rols MP, Delteil C, Golzio M et al. In vivo electrically mediated protein and gene transfer in murine melanoma Nat Biotechnol 1998 16: 168–171

    Article  CAS  Google Scholar 

  30. Nishi T, Yoshizato K, Yamashiro S et al. High-efficiency in vivo gene transfer using intraarterial plasmid DNA injectionfollowing in vivo electroporation Cancer Res 1996 56: 1050–1055

    CAS  PubMed  Google Scholar 

  31. Goto T, Nishi T, Tamura T 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  Google Scholar 

  32. Baba M, Iishi H, Tatsuta M . In vivo electrophoretic transfer of bcl-2 antisense oligonucleotide inhibits the development of hepatocellular carcinoma in rats Int J Cancer 2000 85: 260–266

    Article  CAS  Google Scholar 

  33. Bettan M, Ivanov MA, Mir LM et al. Efficient DNA electrotransfer into tumors Bioelectrochemistry 2000 52: 83–90

    Article  CAS  Google Scholar 

  34. Lohr F, Lo DY, Zaharoff DA et al. Effective tumor therapy with plasmid-encoded cytokines combined with in vivo electroporation Cancer Res 2001 61: 3281–3284

    CAS  PubMed  Google Scholar 

  35. Yoshizato K, Nishi T, Goto T et al. Gene delivery with optimized electroporation parameters shows potential for treatment of gliomas Int J Oncol 2000 16: 899–905

    CAS  PubMed  Google Scholar 

  36. Susil R, Semrov D, Miklavcic D . Electric field induced transmembrane potential depends on cell density and organization Electro-Magnetobiol 1998 17: 391–399

    Article  Google Scholar 

  37. O'Hare MJ, Ormerod MG, Imrie PR et al. Electropermeabilization and electrosensitivity of different types of mammalian cells In: Neumann E, Sowers AE, Jordan CA, eds Electroporation and Electrofusion in Cell Biology New York: Plenum 1989 319–330

    Chapter  Google Scholar 

  38. Cemazar M, Jarm T, Miklavcic D et al. Effect of electric-field intensity on electropermeabilization and electrosensitivity of various tumor-cell lines in vitro Electro-Magnetobiol 1998 17: 261–270

    Google Scholar 

  39. Teissie J, Rols MP . Manipulation of cell cytoskeleton affects the lifetime of cell membrane electropermeabilization Ann NY Acad Sci 1994 720: 98–110

    Article  CAS  Google Scholar 

  40. Meaking WS, Edgerton J, Harton CW et al. Electroporation-induced damage in mammalian cell DNA Biochim Biophys Acta 1995 1264: 357–362

    Article  Google Scholar 

  41. Coralli C, Cemazar M, Kanthou C et al. Limitations of the reporter green fluorescent protein under simulated tumor conditions Cancer Res 2001 61: 4784–4790

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the Cancer Research Campaign, Grant SP2292/0102, and the Ministry of Education, Science, and Sport of the Republic of Slovenia. We would like to thank the Advanced Technology Development Group (Gray Cancer Institute) for the production of the custom-made square wave electroporator and imaging system.

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Authors and Affiliations

  1. Tumor Microcirculation Group, Gray Cancer Institute, Mount Vernon Hospital, Northwood, HA6 2JR, UK

    Maja Cemazar, John Wilson, Gillian M Tozer & Gabi U Dachs

  2. Department of Tumor Biology, Institute of Oncology, Ljubljana, SI-1000, Slovenia

    Maja Cemazar, Gregor Sersa & Alenka Grosel

  3. Molecular Immunology Unit, Institute of Child Health, University College London, London, WC1N 1EH, UK

    Stephen L Hart

Authors
  1. Maja Cemazar
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  2. Gregor Sersa
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  4. Gillian M Tozer
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Correspondence to Maja Cemazar.

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Cemazar, M., Sersa, G., Wilson, J. et al. Effective gene transfer to solid tumors using different nonviral gene delivery techniques: Electroporation, liposomes, and integrin-targeted vector. Cancer Gene Ther 9, 399–406 (2002). https://doi.org/10.1038/sj.cgt.7700454

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