Preclinical evaluation of a recombinant anti-prostate specific membrane antigen single-chain immunotoxin against prostate cancer

J Immunother. 2010 Apr;33(3):262-71. doi: 10.1097/CJI.0b013e3181c5495c.

Abstract

The prostate-specific membrane antigen (PSMA) is abundantly expressed on prostate cancer epithelial cells and its expression correlates with tumor progression. Therefore, a specific immunotherapy against this antigen may be a novel therapeutic option for the management of prostate cancer. We generated an anti-PSMA single-chain antibody fragment (scFv), called D7, by phage display from the monoclonal antibody 3/F11. By C-terminal ligation of the toxic domain of Pseudomonas Exotoxin A (PE40) to the genes of D7, the immunotoxin D7-PE40 was generated. D7 and D7-PE40 specifically bound to PSMA transfectants and to the PSMA expressing prostate cancer cell line C4-2. In addition, D7-PE40 showed a high serum stability and induced a 50% reduction of viability (IC50) in C4-2 cells at a concentration of 140 pM. In vivo, D7-PE40 was well tolerated in SCID mice up to a single dose of 20 microg, whereas higher doses induced severe hepatotoxicity with deaths of the animals. Immunotoxin treatment of mice bearing C4-2 tumor xenografts caused a significant inhibition of tumor growth, whereas mice with PSMA-negative DU 145 tumors remained unaffected. Owing to its high and specific cytotoxicity and its capability to inhibit prostate tumor growth in vivo the immunotoxin D7-PE40 represents a promising candidate for the immunotherapy of prostate cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP Ribose Transferases / genetics
  • ADP Ribose Transferases / immunology
  • Animals
  • Antigens, Surface / genetics
  • Antigens, Surface / immunology*
  • Bacterial Toxins / genetics
  • Bacterial Toxins / immunology
  • Cell Line
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cell Survival / immunology
  • Drug Screening Assays, Antitumor
  • Exotoxins / genetics
  • Exotoxins / immunology
  • Glutamate Carboxypeptidase II / genetics
  • Glutamate Carboxypeptidase II / immunology*
  • Humans
  • Immunotoxins / genetics
  • Immunotoxins / immunology*
  • Immunotoxins / pharmacology
  • Male
  • Mice
  • Mice, SCID
  • Prostatic Neoplasms / immunology*
  • Prostatic Neoplasms / pathology
  • Prostatic Neoplasms / prevention & control
  • Pseudomonas aeruginosa Exotoxin A
  • Recombinant Proteins / immunology
  • Single-Chain Antibodies / genetics
  • Single-Chain Antibodies / immunology*
  • Tumor Burden / drug effects
  • Virulence Factors / genetics
  • Virulence Factors / immunology
  • Xenograft Model Antitumor Assays

Substances

  • Antigens, Surface
  • Bacterial Toxins
  • Exotoxins
  • Immunotoxins
  • Recombinant Proteins
  • Single-Chain Antibodies
  • Virulence Factors
  • ADP Ribose Transferases
  • FOLH1 protein, human
  • Glutamate Carboxypeptidase II