Cancer Immunology, Immunotherapy

, Volume 55, Issue 11, pp 1367–1373 | Cite as

A recombinant PSMA-specific single-chain immunotoxin has potent and selective toxicity against prostate cancer cells

  • P. Wolf
  • D. Gierschner
  • P. Bühler
  • U. Wetterauer
  • U. Elsässer-Beile
Original Article

Abstract

Prostate cancer is the most commonly diagnosed form of cancer and the second leading cancer-related death among men in the Western civilization. Since no effective therapy exists for this tumor after progression beyond resectable boundaries, there is an urgent need for new treatment strategies. Prostate specific membrane antigen (PSMA) represents an excellent target on prostate cancer cells, and therefore specific immunotherapy may be a novel therapeutic option for the management of this tumor. We constructed a fully recombinant immunotoxin (A5-PE40) from a single-chain antibody fragment (scFv) against cell-adherent PSMA and a truncated form of Pseudomonas exotoxin A (PE40) lacking its natural binding domain Ia. The scFv A5 was obtained from a mAb elicited with native PSMA by phage display technology and direct selection on cells carrying the antigen. The bacterially expressed and purified immunotoxin A5-PE40 specifically binds to PSMA-positive prostate cancer cells and induces a 50% reduction of viability (IC50) at a concentration of 20 pM, while PSMA-negative cells remain unaffected. Due to its high and specific toxicity this recombinant immunotoxin is a promising candidate for therapeutic applications in patients with prostate cancer.

Keywords

Prostate cancer Recombinant immunotoxin Immunotherapy PSMA 

References

  1. 1.
    Bander NH, Trabulsi EJ, Kostakoglu L, Yao D, Vallabhajosula S, Smith-Jones P, Joyce MA, Milowsky M, Nanus DM, Goldsmith SJ (2003) Targeting metastatic prostate cancer with radiolabeled monoclonal antibody J591 to the extracellular domain of prostate specific membrane antigen. J Urol 170:1717–1721PubMedCrossRefGoogle Scholar
  2. 2.
    Chang SS, Reuter VE, Heston WD, Gaudin PB (2001) Comparison of anti-prostate-specific membrane antigen antibodies and other immunomarkers in metastatic prostate carcinoma. Urology 57:1179–1183PubMedCrossRefGoogle Scholar
  3. 3.
    Chiarodo A (1991) National Cancer Institute roundtable on prostate cancer: future research directions. Cancer Res 51:2498–2505PubMedGoogle Scholar
  4. 4.
    Davis MI, Bennett MJ, Thomas LM, Bjorkman PJ (2005) Crystal structure of prostate-specific membrane antigen, a tumor marker and peptidase. Proc Natl Acad Sci USA 102:5981–5986PubMedCrossRefGoogle Scholar
  5. 5.
    Di Paolo C, Willuda J, Kubetzko S, Lauffer I, Tschudi D, Waibel R, Pluckthun A, Stahel RA, Zangemeister-Wittke U (2003) A recombinant immunotoxin derived from a humanized epithelial cell adhesion molecule-specific single-chain antibody fragment has potent and selective antitumor activity. Clin Cancer Res 9:2837–2848PubMedGoogle Scholar
  6. 6.
    Fracasso G, Bellisola G, Cingarlini S, Castelletti D, Prayer-Galetti T, Pagano F, Tridente G, Colombatti M (2002) Anti-tumor effects of toxins targeted to the prostate specific membrane antigen. Prostate 53:9–23PubMedCrossRefGoogle Scholar
  7. 7.
    Fuchs P, Dubel S, Breitling F, Braunagel M, Klewinghaus I, Little M (1992) Recombinant human monoclonal antibodies. Basic principles of the immune system transferred to E. coli. Cell Biophys 21:81–91PubMedGoogle Scholar
  8. 8.
    Ghosh A, Heston WD (2004) Tumor target prostate specific membrane antigen (PSMA) and its regulation in prostate cancer. J Cell Biochem 91:528–539PubMedCrossRefGoogle Scholar
  9. 9.
    Huang X, Bennett M, Thorpe PE (2004) Anti-tumor effects and lack of side effects in mice of an immunotoxin directed against human and mouse prostate-specific membrane antigen. Prostate 61:1–11PubMedCrossRefGoogle Scholar
  10. 10.
    Hwang J, Fitzgerald DJ, Adhya S, Pastan I (1987) Functional domains of Pseudomonas exotoxin identified by deletion analysis of the gene expressed in E. coli. Cell 48:129–136PubMedCrossRefGoogle Scholar
  11. 11.
    Israeli RS, Powell CT, Corr JG, Fair WR, Heston WD (1994) Expression of the prostate-specific membrane antigen. Cancer Res 54:1807–1811PubMedGoogle Scholar
  12. 12.
    Jemal A, Tiwari RC, Murray T, Ghafoor A, Samuels A, Ward E, Feuer EJ, Thun MJ (2004) Cancer statistics, 2004. CA Cancer J Clin 54:8–29PubMedCrossRefGoogle Scholar
  13. 13.
    Kawakami M, Nakayama J (1997) Enhanced expression of prostate-specific membrane antigen gene in prostate cancer as revealed by in situ hybridization. Cancer Res 57:2321–2324PubMedGoogle Scholar
  14. 14.
    Kipriyanov SM, Moldenhauer G, Little M (1997) High level production of soluble single chain antibodies in small-scale Escherichia coli cultures. J Immunol Methods 200:69–77PubMedCrossRefGoogle Scholar
  15. 15.
    Klimka A, Barth S, Matthey B, Roovers RC, Lemke H, Hansen H, Arends JW, Diehl V, Hoogenboom HR, Engert A (1999) An anti-CD30 single-chain Fv selected by phage display and fused to Pseudomonas exotoxin A (Ki-4(scFv)-ETA′) is a potent immunotoxin against a Hodgkin-derived cell line. Br J Cancer 80:1214–1222PubMedCrossRefGoogle Scholar
  16. 16.
    Liu H, Moy P, Kim S, Xia Y, Rajasekaran A, Navarro V, Knudsen B, Bander NH (1997) Monoclonal antibodies to the extracellular domain of prostate-specific membrane antigen also react with tumor vascular endothelium. Cancer Res 57:3629–3634PubMedGoogle Scholar
  17. 17.
    Liu H, Rajasekaran AK, Moy P, Xia Y, Kim S, Navarro V, Rahmati R, Bander NH (1998) Constitutive and antibody-induced internalization of prostate-specific membrane antigen. Cancer Res 58:4055–4060PubMedGoogle Scholar
  18. 18.
    MacDonald GC, Glover N (2005) Effective tumor targeting: strategies for the delivery of armed antibodies. Curr Opin Drug Discov Devel 8:177–183PubMedGoogle Scholar
  19. 19.
    Marks JD, Hoogenboom HR, Bonnert TP, McCafferty J, Griffiths AD, Winter G (1991) By-passing immunization. Human antibodies from V-gene libraries displayed on phage. J Mol Biol 222:581–597PubMedCrossRefGoogle Scholar
  20. 20.
    Orum H, Andersen PS, Oster A, Johansen LK, Riise E, Bjornvad M, Svendsen I, Engberg J (1993) Efficient method for constructing comprehensive murine Fab antibody libraries displayed on phage. Nucleic Acids Res 21:4491–4498PubMedCrossRefGoogle Scholar
  21. 21.
    Patri AK, Myc A, Beals J, Thomas TP, Bander NH, Baker JR Jr (2004) Synthesis and in vitro testing of J591 antibody-dendrimer conjugates for targeted prostate cancer therapy. Bioconjug Chem 15:1174–1181PubMedCrossRefGoogle Scholar
  22. 22.
    Peipp M, Kupers H, Saul D, Schlierf B, Greil J, Zunino SJ, Gramatzki M, Fey GH (2002) A recombinant CD7-specific single-chain immunotoxin is a potent inducer of apoptosis in acute leukemic T cells. Cancer Res 62:2848–2855PubMedGoogle Scholar
  23. 23.
    Schulke N, Varlamova OA, Donovan GP, Ma D, Gardner JP, Morrissey DM, Arrigale RR, Zhan C, Chodera AJ, Surowitz KG, Maddon PJ, Heston WD, Olson WC (2003) The homodimer of prostate-specific membrane antigen is a functional target for cancer therapy. Proc Natl Acad Sci USA 100:12590–12595PubMedCrossRefGoogle Scholar
  24. 24.
    Silver DA, Pellicer I, Fair WR, Heston WD, Cordon-Cardo C (1997) Prostate-specific membrane antigen expression in normal and malignant human tissues. Clin Cancer Res 3:81–85PubMedGoogle Scholar
  25. 25.
    Wang S, Diamond DL, Hass GM, Sokoloff R, Vessella RL (2001) Identification of prostate specific membrane antigen (PSMA) as the target of monoclonal antibody 107-1A4 by proteinchip; array, surface-enhanced laser desorption/ionization (SELDI) technology. Int J Cancer 92:871–876PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • P. Wolf
    • 1
    • 2
  • D. Gierschner
    • 1
  • P. Bühler
    • 1
    • 2
  • U. Wetterauer
    • 1
  • U. Elsässer-Beile
    • 1
  1. 1.Department of Urology, Experimental UrologyUniversity of FreiburgFreiburgGermany
  2. 2.Faculty of BiologyUniversity of FreiburgFreiburgGermany

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