Pro-chemotherapeutic effects of antibody against extracellular domain of claudin-4 in bladder cancer.
Bladder cancer displays an aggressive phenotype in the muscle-invasive phase, and is associated with a high mortality rate. Therefore, novel molecular therapeutic targets are needed to improve patient survival. A monoclonal antibody against the extracellular domain of the claudin-4 (CLDN4) tight junction protein was established by immunizing rats with a plasmid vector encoding human CLDN4. A hybridoma clone, producing a rat monoclonal antibody recognizing CLDN4 (clone 4D3), was obtained. Immunohistochemistry by using the 4D3 antibody showed that CLDN4 expression was associated with local invasion, nodal metastasis, distant metastasis, and advanced stage in 86 cases of bladder cancer. The 4D3 antibody inhibited growth, invasion, and survival, associated with abrogation of the intratumoral microenvironment; lowered concentrations of epidermal growth factor and vascular endothelial growth factor were found in three-dimensional cultures of T24 and RT4 cells. In combination with cisplatin therapy, 4D3 enhanced cisplatin cytotoxicity by increasing cellular permeability, leading to increased intracellular cisplatin concentrations. In mouse models of subcutaneous tumors and lung metastasis, 4D3 enhanced tumor growth inhibition, alone and with concurrent cisplatin treatment. The anti-tumor activity of the newly established 4D3 antibody suggests that it may be a powerful tool in CLDN4-targeting therapy, and in combination with chemotherapy.
Department of Molecular Pathology, Nara Medical University, Nara 634-8521, Japan; Department of Urology, Nara Medical University, Nara 634-8521, Japan.
Masaomi
M
Kuwada
Department of Urology, Nara Medical University, Nara 634-8521, Japan.
Yoshitomo
Y
Chihara
Department of Molecular Pathology, Nara Medical University, Nara 634-8521, Japan.
Yi
Y
Luo
Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
Xiangru
X
Li
Department of Molecular Pathology, Nara Medical University, Nara 634-8521, Japan.
Yukiko
Y
Nishiguchi
Department of Molecular Pathology, Nara Medical University, Nara 634-8521, Japan.
Rina
R
Fujiwara
Department of Gastrointestinal Surgery, Fukuoka University School of Medicine, Fukuoka 7-45-1, Japan.
Takamistu
T
Sasaki
Department of Molecular Pathology, Nara Medical University, Nara 634-8521, Japan.
Kiyomu
K
Fujii
Department of Molecular Pathology, Nara Medical University, Nara 634-8521, Japan.
Hitoshi
H
Ohmori
Department of Urology, Nara Medical University, Nara 634-8521, Japan.
Kiyohide
K
Fujimoto
Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
Masuo
M
Kondoh
Department of Molecular Pathology, Nara Medical University, Nara 634-8521, Japan. Electronic address: cooninh@zb4.so-net.ne.jp.
Hiroki
H
Kuniyasu
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology / Q000627:therapeutic use*; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000008:administration & dosage; D062506:Claudin-4 / Q000276:immunology*; D004357:Drug Synergism; D005260:Female; D006801:Humans; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008875:Middle Aged; D017208:Rats, Wistar; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy* / Q000473:pathology; D023041:Xenograft Model Antitumor Assays
D000911:Antibodies, Monoclonal; C568157:CLDN4 protein, human; D062506:Claudin-4; D002945:Cisplatin
Antibody therapy; Claudin; Microenvironment; Tight junction