Biotechnological and gene therapeutic strategies in cancer treatment.
New anti-cancer agents are being developed which incorporate cancer-cell-specific recognition functions and are thus able to distinguish between normal and tumor cells. Recognition is dependent on the enhanced expression of antigenic determinants on the surface of tumor cells. The ErbB-2 receptor (ErbB-2R) is overproduced in a high percentage of adenocarcinomas arising in the breast, ovary, lung and stomach, when compared to normal cells. The tumor-enriched expression and extracellular accessibility make this receptor a suitable target for directed tumor therapy. A gene expressing the single-chain antibody molecule (scFv), specific for the extracellular domain of the ErbB-2R, was constructed by joining cDNAs encoding the light- and heavy-chain variable domains of the monoclonal antibody (mAb) FRP5. This scFv-encoding gene has been used as a targeting domain for two effectors: (i) A recombinant immunotoxin-encoding gene was constructed by adding sequences encoding a modified Pseudomonas aeroginosa exotoxin A (ETA) to the scFv-encoding DNA. (ii) Cytotoxic T-lymphocytes (CTL) with specificity for ErbB-2R-producing tumor cells were generated by retroviral transfer of a chimeric gene which encodes the scFv(FRP5), a hinge region and the zeta-chain of the T-cell receptor (TCR) complex. The bacterially produced recombinant immunotoxin scFv(FRP5)-ETA binds specifically to the ErbB-2R and displays both in vitro and in vivo cytotoxic effects selective for tumor cells producing high levels of the ErbB-2R. Target cells expressing the ErbB-2R gene were lysed in vitro with high specificity by the scFv::hinge::zeta-expressing T-cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Wels
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Institute for Experimental Cancer Research, Tumor Biology Center, Freiburg, Germany.
Moritz
D
D
Schmidt
M
M
Jeschke
M
M
Hynes
N E
NE
Groner
B
B
D016475:3T3 Cells; D036002:ADP Ribose Transferases; D000595:Amino Acid Sequence; D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics / Q000627:therapeutic use; D001483:Base Sequence; D002461:Cell Line, Transformed; D005098:Exotoxins / Q000235:genetics / Q000627:therapeutic use*; D018014:Gene Transfer Techniques; D005803:Genes, Immunoglobulin / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics; D000922:Immunotoxins / Q000235:genetics / Q000627:therapeutic use*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008969:Molecular Sequence Data; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000628:therapy*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics* / Q000276:immunology; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D001427:Bacterial Toxins; D005098:Exotoxins; D007135:Immunoglobulin Variable Region; D000922:Immunotoxins; D018719:Erb-b2 Receptor Tyrosine Kinases; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A