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Anti?EpCAM monoclonal antibody exerts antitumor activity against oral squamous cell carcinomas.

ABSTRACT: The epithelial cell adhesion molecule (EpCAM) is a calcium?independent, homophilic, intercellular adhesion factor classified as a transmembrane glycoprotein. In addition to cell adhesion, EpCAM also contributes to cell signaling, differentiation, proliferation, and migration. EpCAM is an essential factor in the carcinogenesis of numerous human cancers. In the present study, we developed and validated an anti?EpCAM monoclonal antibody (mAb), EpMab?16 (IgG2a, kappa), by immunizing mice with EpCAM?overexpressing CHO?K1 cells. EpMab?16 specifically reacted with endogenous EpCAM in oral squamous cell carcinoma (OSCC) cell lines in flow cytometry and Western blot analyses. It exhibited a plasma membrane?like stain pattern in OSCC tissues upon immunohistochemical analysis. The KD for EpMab?16 in SAS and HSC?2 OSCC cells were assessed via flow cytometry at 1.1x10?8 and 1.9x10?8 M, respectively, suggesting moderate binding affinity of EpMab?16 for EpCAM. We then assessed whether the EpMab?16 induced antibody?dependent cellular cytotoxicity (ADCC) and complement?dependent cytotoxicity (CDC) against OSCC cell lines, and antitumor capacity in a murine xenograft model. In vitro experiments revealed strong ADCC and CDC inducement against OSCC cells treated with EpMab?16. In vivo experiments on OSCC xenografts revealed that EpMab?16 treatment significantly reduced tumor growth compared with the control mouse IgG. These data indicated that EpMab?16 could be a promising treatment option for EpCAM?expressing OSCCs.

SUBMITTER: Kaneko MK

PROVIDER: S-EPMC7640354 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Anti‑EpCAM monoclonal antibody exerts antitumor activity against oral squamous cell carcinomas.

Kaneko Mika K MK Ohishi Tomokazu T Takei Junko J Sano Masato M Nakamura Takuro T Hosono Hideki H Yanaka Miyuki M Asano Teizo T Sayama Yusuke Y Harada Hiroyuki H Kawada Manabu M Kato Yukinari Y

Oncology reports 20201013 6

The epithelial cell adhesion molecule (EpCAM) is a calcium‑independent, homophilic, intercellular adhesion factor classified as a transmembrane glycoprotein. In addition to cell adhesion, EpCAM also contributes to cell signaling, differentiation, proliferation, and migration. EpCAM is an essential factor in the carcinogenesis of numerous human cancers. In the present study, we developed and validated an anti‑EpCAM monoclonal antibody (mAb), EpMab‑16 (IgG2a, kappa), by immunizing mice with EpCAM‑ ...[more]

PMID: 33125138

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Project description:CD44 is widely expressed on the surface of most tissues and all hematopoietic cells, and regulates many genes associated with cell adhesion, migration, proliferation, differentiation, and survival. CD44 has also been studied as a therapeutic target in several cancers. Previously, an anti‑CD44 monoclonal antibody (mAb), C44Mab‑5 (IgG1, kappa) was established by immunizing mice with CD44‑overexpressing Chinese hamster ovary (CHO)-K1 cells. C44Mab‑5 recognized all CD44 isoforms, and showed high sensitivity for flow cytometry and immunohistochemical analysis in oral cancers. However, as the IgG1 subclass of C44Mab‑5 lacks antibody‑dependent cellular cytotoxicity (ADCC) and complement‑dependent cytotoxicity (CDC), the antitumor activity of C44Mab‑5 could not be determined. In the present study, we converted the mouse IgG1 subclass antibody C44Mab‑5 into an IgG2a subclass antibody, 5‑mG2a, and further produced a defucosylated version, 5‑mG2a‑f, using FUT8‑deficient ExpiCHO‑S (BINDS‑09) cells. Defucosylation of 5‑mG2a‑f was confirmed using fucose‑binding lectins, such as AAL and PhoSL. The dissociation constants (KD) for 5‑mG2a‑f against SAS and HSC‑2 oral cancer cells were determined through flow cytometry to be 2.8x10‑10 M and 2.6x10‑9 M, respectively, indicating that 5‑mG2a‑f possesses extremely high binding affinity. Furthermore, immunohistochemical staining using 5‑mG2a‑f specifically stained the membranes of oral cancer cells. In vitro analysis demonstrated that 5‑mG2a‑f showed moderate ADCC and CDC activities against SAS and HSC‑2 oral cancer cells. In vivo analysis revealed that 5‑mG2a‑f significantly reduced tumor development in SAS and HSC‑2 xenografts in comparison to control mouse IgG, even after injection seven days post‑tumor inoculation. Collectively, these results suggest that treatment with 5‑mG2a‑f may represent a useful therapy for patients with CD44‑expressing oral cancers.

Dataset Information

Anti?EpCAM monoclonal antibody exerts antitumor activity against oral squamous cell carcinomas.

Publications

Anti‑EpCAM monoclonal antibody exerts antitumor activity against oral squamous cell carcinomas.

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