Engineering and biological characterization of VB6-845, an anti-EpCAM immunotoxin containing a T-cell epitope-depleted variant of the plant toxin bouganin.
The clinical development of immunotoxins in the treatment of solid tumors has been impeded in part, by the induction of an immune response directed primarily against the toxin moiety. Bouganin, a type I ribosome inactivating protein isolated from the leaf of Bougainvillea spectabilis Willd, was mutated to remove the T-cell epitopes while preserving the biological activity of the wild-type molecule. The T-cell epitope-depleted variant of bouganin (de-bouganin) was genetically linked to an anti-epithelial cell adhesion molecule (EpCAM) Fab moiety via a peptidic linker containing a furin proteolytic site to create the fusion construct VB6-845. To determine the optimal construct design for VB6-845, several dicistronic units where de-bouganin was genetically linked to either the N-terminal or C-terminal of either the heavy or light chain were engineered. Only the C-terminal variants expressed the full-length molecule. An in vitro assessment of the biological activity of VB6-845 showed that it bound and selectively killed EpCAM-positive cell lines with a greater potency than many commonly used chemotherapeutic agents. In vivo efficacy was demonstrated using an EpCAM-positive human tumor xenograft model in SCID mice with the majority of the mice treated being tumor free at the end of the study.
Cizeau
Jeannick
J
Viventia Biotechnologies Inc, Manitoba, Canada.
Grenkow
Danielle M
DM
Brown
Jennifer G
JG
Entwistle
Joycelyn
J
MacDonald
Glen C
GC
D000595:Amino Acid Sequence; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000276:immunology / Q000627:therapeutic use*; D015815:Cell Adhesion Molecules / Q000276:immunology* / Q000378:metabolism; D045744:Cell Line, Tumor; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology; D005260:Female; D006801:Humans; D000922:Immunotoxins / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000188:drug therapy*; D039281:Nyctaginaceae / Q000235:genetics / Q000276:immunology; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D016415:Sequence Alignment; D023041:Xenograft Model Antitumor Assays
D000972:Antineoplastic Agents, Phytogenic; D015815:Cell Adhesion Molecules; D018984:Epitopes, T-Lymphocyte; D000922:Immunotoxins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C544011:VB6-845 fusion protein