Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

A two-step targeting approach for delivery of doxorubicin-loaded liposomes to tumour cells in vivo

Abstract

A two-step targeting approach was used to deliver doxorubicin-loaded liposomes to a murine tumour cell (P388 leukaemia) grown in culture and, more importantly, in vivo. Targeting was mediated through the use of an antibody specific for the Thy 1.2 antigen that is highly expressed on P388 cells. Briefly, the approach consists of prelabeling target cells with biotinylated anti-Thy 1.2 antibody prior to administration of drug-loaded liposomes that have streptavidin covalently attached to their surface. Results from in vitro studies demonstrate that a 30-fold increase in cell-associated lipid and a 20-fold increase in cell-associated doxorubicin can be achieved over control liposomes using this two-step procedure. Flow-cytometry and fluorescent-microscopy data were used to confirm that P388 cells can be stably labeled with the biotinylated anti-Thy 1.2 antibody in vivo. Subsequently, liposome-targeting studies were initiated in vivo, where target cell binding was assessed following i.p. or i.v. injection of doxorubicinloaded liposomes into animals bearing P388 tumours prelabeled with biotinylated antibody. A streptavidin-mediated 3.7-fold increase in cell-associated lipid and drug was achieved when the liposomes were given i.p. When doxorubicin-loaded streptavidin liposomes were injected i.v., P388 cells located in the peritoneal cavity were specifically labeled, although the efficiency of this targeting reaction was low. Less than a 2-fold increase in cell-associated lipid was achieved through the use of target-specific (streptavidin-coated) liposomes. These studies demonstrate that the presence of a well-labeled target cell population within the peritoneal cavity will not promote accumulation of an i.v. injected, targeted liposomal drug. Furthermore, the importance of separating target-cell-specific binding from non-specific uptake by tumour-associated macrophages is discussed.

This is a preview of subscription content, log in to check access.

References

  1. 1.

    Ahmad I, Allen TM (1992) Antibody-mediated specific binding and cytotoxicity of liposome-entrapped doxorubicin to lung cancer cells in vitro. Cancer Res 52:4817–4820

  2. 2.

    Ahmad I, Longenecker M, Samuel J, Allen TM (1993) Antibody-targeted delivery of doxorubicin entrapped in sterically stabilized liposomes can eradicate lung cancer in mice. Cancer Res 53: 1484–1488

  3. 3.

    Allen TM (1989) Stealth liposomes: avoiding reticuloendothelial uptake. In: Lopez-Berestein G, Fidler IJ (eds) Liposomes in the therapy of infectious diseases and cancer. Alan R. Liss, New York, pp 405–415

  4. 4.

    Aragnol D, Leserman LD (1986) Immune clearance of liposomes inhibited by an anti-Fc receptor antibody in vivo. Proc Natl Acad Sci USA 83:2699–2703

  5. 5.

    Balazsovits JAE, Mayer LD, Bally MB, Cullis PR, McDonell M, Ginsberg RS, Falk RE (1989) Analysis of the effect of liposome encapsulation on the vesicant properties, acute and cardiac toxicities, and antitumor efficacy of doxorubicin. Cancer Chemother Pharmacol 23:81–86

  6. 6.

    Bally MB, Hope MJ, Mayer LD, Madden TD, Cullis PR (1988) Novel procedures for generating and loading liposomal systems. In: Gregoriadis G (ed) Liposomes as drug carriers. John Wiley and Sons, Chichester, pp 841–853

  7. 7.

    Bally MB, Nayar R, Masin D, Hope MJ, Cullis PR, Mayer LD (1990) Liposomes with entrapped doxorubicin exhibit extended blood residence times. Biochim Biophys Acta 1023:133–139

  8. 8.

    Bally MB, Masin D, Nayar R, Cullis PR, Mayer LD (1994) Transfer of liposomal drug carriers from the blood to the peritoneal cavity of normal and ascitic tumor-bearing mice. Cancer Chemother Pharmacol 34:137–146

  9. 9.

    Bankert RB, Yokota S, Ghosh SK, Mayhew E, Jou Y-H (1989) Immunospecific targeting of cytosine arabinonucleoside-containing liposomes to the idiotype on the surface of a murine B-cell tumor in vitro and in vivo. Cancer Res 49:301–308

  10. 10.

    Carlsson J, Drevin H, Axen R (1978) Protein thiolation and reversible protein-protein conjugation,N-succinimidyl 3-(2-pyri-dyldithio) propionate, a new heterobifunctional reagent. Biochem J 173:723–737

  11. 11.

    Debs RJ, Heath TD, Papahadjopoulos D (1987) Targeting of anti-Thy 1.1 monoclonal antibody conjugated liposomes in Thy 1.1 mice after intravenous administration. Biochim Biophys Acta 901: 183–190

  12. 12.

    Gabizon AA (1992) Selective tumor localization and improved therapeutic index of anthracyclines encapsulated in long-circulating liposomes. Cancer Res 52:891–896

  13. 13.

    Gabizon A, Dagan A, Goren D, Barenholz Y, Fuks Z (1982) Liposomes as in vivo carriers of adriamycin: reduced cardiac uptake and preserved antitumor activity in mice. Cancer Res 42: 4734–4739

  14. 14.

    Gabizon A, Goren D, Fuks Z, Meshoren A, Barenholz Y (1985) Superior therapeutic activity of liposome-associated adriamycin in a murine metastatic tumour model. Br J Cancer 51:681–689

  15. 15.

    Gabizon A, Meshozer A, Barenholz Y (1986) Comparative longterm study of the toxicities of free and liposome-associated doxorubicin in mice after intravenous administration. J Natl Cancer Inst 77:459–467

  16. 16.

    Heath TD, Montgomery JA, Piper JR, Papahadjopoulos D (1983) Antibody-targeted liposomes: increase in specific toxicity of methotrexate-γ-aspartate. Proc Natl Acad Sci USA 80:1377–1381

  17. 17.

    Hope MJ, Bally MB, Webb G, Cullis PR (1985) Production of large unilamellar vesicles by a rapid extrusion procedure. Characterization of size distribution, trapped volume and ability to maintain a membrane potential. Biochim Biophys Acta 812: 55–65

  18. 18.

    Huang SK, Mayhew E, Gilani S, Lasic DD, Martin FJ, Papahadjopoulos D (1992) Pharmacokinetics and therapeutics of sterically stabilized liposomes in mice bearing C-26 colon carcinoma. Cancer Res 52:6774–6781

  19. 19.

    Konno H, Tadakuma T, Kumai K, Takahashi T, Ishibiki K, Abe O, Sakaguchi S (1990) The antitumor effects of adriamycin entrapped in liposomes on lymph node metastases. Jpn J Surg 20:424–428

  20. 20.

    Ledbetter JA, Herzenberg LA (1979) Xenogeneic monoclonal antibodies to mouse lymphoid differentiation antigens. Immunol Rev 47:63–90

  21. 21.

    Leserman LD, Machy P, Barbet J (1981) Cell-specific drug transfer from liposomes bearing monoclonal antibodies. Nature 293:226–228

  22. 22.

    Loughrey H, Bally MB, Cullis PR (1987) A non-covalent method of attaching antibodies to liposomes. Biochim Biophys Acta 901: 157–160

  23. 23.

    Loughrey HC, Choi LS, Cullis PR, Bally MB (1990) Optimized procedures for the coupling of proteins to liposomes. J Immunol Methods 132:25–35

  24. 24.

    Loughrey HC, Wong KF, Choi LS, Cullis PR, Bally MB (1990) Protein-liposome conjugates with defined size distributions. Biochim Biophys Acta 1028:73–81

  25. 25.

    Loughrey HC, Choi LS, Wong KF, Cullis PR, Bally MB (1993) Preparation of streptavidin-liposomes for use in ligand-specific targeting applications. In: Gregoriadis G (ed) Liposome technology, vol 3. CRC, Boca Raton, Florida, pp 163–178

  26. 26.

    Matthay KK, Heath TD, Papahadjopoulos D (1984) Specific enhancement of drug delivery to AKR lymphoma by antibody-targeted small unilamellar vesicles. Cancer Res 44:1880–1886

  27. 27.

    Mayer LD, Bally MB, Cullis PR (1990) Strategies of optimizing liposomal doxorubicin. J Liposome Res 4:463–480

  28. 28.

    Mayer LD, Bally MB, Cullis PR, Wilson SL, Emerman JT (1990) Comparison of free and liposome encapsulated doxorubicin tumor drug uptake and antitumor efficacy in the SC115 murine mammary tumor. Cancer Lett 53:183–189

  29. 29.

    Nässander UK, Steerenberg PA, Poppe H, Storm G, Poels LG, De Jong WH, Crommelin DJA (1992) In vivo targeting of OV-TL 3 immunoliposomes to ascitic ovarian carcinoma cells (OVCAR-3) in athymic nude mice. Cancer Res 52:646–653

  30. 30.

    Olson F, Mayhew E, Maslow D, Rustum Y, Szoka F (1982) Characterization, toxicity and therapeutic efficacy of adriamycin encapsulated in liposomes. Eur J Cancer Clin Oncol 18:167–176

  31. 31.

    Papahadjopoulos D, Allen TM, Gabizon A, Mayhew E, Matthay K, Huang SK, Lee KD, Woodle MC, Lasic DD, Redemann C, Martin FJ (1992) Sterically stabilized liposomes: improvements in pharmacokinetics and antitumor therapeutic efficacy. Proc Natl Acad Sci USA 88:11460–11464

  32. 32.

    Porstmann B, Porstmann T, Nugel E, Evers U (1985) Which of the commonly used marker enzymes gives the best results in colorimetric and fluorimetric enzyme immunoassays: horseradish peroxidase, alkaline phosphatase or β-galactosidase? J Immunol Methods 79:27–37

  33. 33.

    Rahman A, Kessler A, More N, Sikic B, Rowden G, Woolley P, Schein PS (1980) Liposomal protection of adriamycin-induced cardiotoxicity in mice. Cancer Res 40:1532–1537

  34. 34.

    Ross DD, Joneckis CC, Ordonez JV, Sisk AM, Wu RK, Hamburger AW, Nora RE (1989) Estimation of cell survival by flow cytometric quantification of fluorescein diacetate/propidium iodide viable cell number. Cancer Res 49:3776–3782

  35. 35.

    Senior J, Crowley JCW, Gregoriadis G (1985) Tissue distribution of liposomes exhibiting long half lives in the circulation after intravenous injection. Biochim Biophys Acta 839:1–8

  36. 36.

    Van Hossel QGCM, Stearenberg PA, Crommelin DJA, Dijk A van, Oost W van, Klein S, Douze JMC, Wildt DJ de, Hillen FC (1984) Reduced cardiotoxicity and nephrotoxicity with preservation of antitumor activity of doxorubicin entrapped in stable liposomes in the LOU/M Wsl rat. Cancer Res 44:3698–3705

Download references

Author information

Correspondence to Marcel B. Bally.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Longman, S.A., Cullis, P.R., Choi, L. et al. A two-step targeting approach for delivery of doxorubicin-loaded liposomes to tumour cells in vivo. Cancer Chemother. Pharmacol. 36, 91–101 (1995). https://doi.org/10.1007/BF00689191

Download citation

Key words

  • Liposomes
  • Targeting
  • Doxorubicin