title: EpCAM- and EGFR-targeted selective gene therapy for biliary cancers using Z33-fiber-modified adenovirus.
abstract: A critical issue in adenovirus (Ad)-based cancer gene therapy is to improve the specificity of gene delivery to cancer cells for better efficacy and safety. We explored methods of retargeting Ad vectors for selective gene therapy of human biliary cancers using the Ad incorporating an IgG Fc-binding motif (Z33) from the Staphylococcus protein A (Ad-FZ33) combined with tumor-specific antibodies. Flow cytometry analysis revealed high-expression levels of epithelial cell adhesion molecule (EpCAM) and epidermal growth factor receptor (EGFR) on human biliary cancer cells. Ad-FZ33 expressing LacZ combined with antibodies against EpCAM or EGFR, followed by β-gal assay, demonstrated highly efficient gene transduction in these biliary cancer cells, compared to the treatment with control antibody or without antibody. Ad-FZ33 expressing uracil phosphoribosyl transferase (UPRT), an enzyme which greatly enhances the toxicity of 5-fluorouracil (FU), combined with antibodies against EpCAM or EGFR, remarkably enhanced the sensitivity of biliary cancer cells to 5-FU. By contrast, the treatment did not affect the 5-FU sensitivity of the cells not expressing EpCAM or EGFR including normal hepatocytes. Finally, treatments with the UPRT-expressing Ad-FZ33 with antibodies against EpCAM or EGFR, followed by 5-FU administration, significantly suppressed the growth of biliary cancer xenografts in nude mice. These results indicate that the gene therapy mediated by the Z33 fiber modified Ad with anti-EpCAM or anti-EGFR antibodies offers a potentially effective therapeutic modality against biliary cancers.
authors.affiliation: Division of Gastroenterology, University of Tsukuba Graduate School of Comprehensive Human Sciences, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Rei
authors.initials: R
authors.lastname: Kawashima
authors.forename: Masato
authors.initials: M
authors.lastname: Abei
authors.forename: Kuniaki
authors.initials: K
authors.lastname: Fukuda
authors.forename: Kiminori
authors.initials: K
authors.lastname: Nakamura
authors.forename: Takehide
authors.initials: T
authors.lastname: Murata
authors.forename: Mariko
authors.initials: M
authors.lastname: Wakayama
authors.forename: Emiko
authors.initials: E
authors.lastname: Seo
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Hasegawa
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Mizuguchi
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Obata
authors.forename: Ichinosuke
authors.initials: I
authors.lastname: Hyodo
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Hamada
authors.forename: Kazunari K
authors.initials: KK
authors.lastname: Yokoyama
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000627:therapeutic use; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000276:immunology; D000964:Antimetabolites, Antineoplastic / Q000627:therapeutic use; D001661:Biliary Tract Neoplasms / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D015153:Blotting, Western; D015815:Cell Adhesion Molecules / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000276:immunology; D003131:Combined Modality Therapy; D000071858:Epithelial Cell Adhesion Molecule; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D005472:Fluorouracil / Q000627:therapeutic use; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000627:therapeutic use; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D007074:Immunoglobulin G / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013205:Staphylococcal Protein A / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000964:Antimetabolites, Antineoplastic; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D012333:RNA, Messenger; D013205:Staphylococcal Protein A; C512478:EGFR protein, human; D066246:ErbB Receptors; D005472:Fluorouracil