Novel human single-domain antibodies exert potent anti-tumor activity by targeting EGF-like repeat epitope of EpCAM

IntroductionEpCAM (Epithelial cell adhesion molecule) is a key cancer stem cell marker involved in cancer progression, making it an important target for both diagnosis and therapy. Despite efforts using anti-EpCAM monoclonal antibodies (mAbs), their anti-tumor effects have been limited. Single-domai...

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Main Authors: Xiaofeng Zhou, Zhifang Liu, Weixiong Zhang, Lin Dai, Tao Chen, Zexiong Lin, Hong Pan, Qi Qi, Henry Wei
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-02-01
Series:Frontiers in Pharmacology
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Online Access:https://www.frontiersin.org/articles/10.3389/fphar.2025.1530268/full
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author Xiaofeng Zhou
Zhifang Liu
Weixiong Zhang
Lin Dai
Tao Chen
Zexiong Lin
Hong Pan
Qi Qi
Henry Wei
author_facet Xiaofeng Zhou
Zhifang Liu
Weixiong Zhang
Lin Dai
Tao Chen
Zexiong Lin
Hong Pan
Qi Qi
Henry Wei
author_sort Xiaofeng Zhou
collection DOAJ
description IntroductionEpCAM (Epithelial cell adhesion molecule) is a key cancer stem cell marker involved in cancer progression, making it an important target for both diagnosis and therapy. Despite efforts using anti-EpCAM monoclonal antibodies (mAbs), their anti-tumor effects have been limited. Single-domain antibodies (sdAbs), in contrast, offer advantages such as efficient tumor penetration and reduced immunogenicity. This study aims to screen and explore novel sdAbs targeting EpCAM for cancer therapy.MethodsA critical EGF-like repeat epitope on the EpCAM extracellular domain was selected for screening a human sdAb library via phage display. The selected sdAbs were purified and their anti-cancer activity was validated through specific binding with the EpCAM peptide. The effects of these sdAbs on cell proliferation, migration, invasion, and apoptosis were tested in vitro, and their anti-tumor activity was assessed in a xenograft model.ResultsFive fully human anti-EpCAM sdAbs were isolated, all of which specifically bound to the EpCAM peptide and showed selective binding to various cancer cell lines, but not to 293T and 3T3 cells. Functional assays demonstrated that these sdAbs significantly inhibited cancer cell proliferation, migration, and invasion, and induced apoptosis. Notably, two sdAbs (aEP3D4 and aEP4G2) exhibited potent anti-tumor effects in vivo, significantly reducing tumor volume and weight in a mouse xenograft model.DiscussionThis study provides compelling evidence that targeting EpCAM with sdAbs is a promising approach for cancer treatment. The identified anti-EpCAM sdAbs exhibit substantial anti-tumor activity both in vitro and in vivo, suggesting they are strong candidates for future therapeutic applications in cancer therapy.
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spelling doaj-art-b623e5459fb442279c695cc249663bbc2025-08-20T03:12:43ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122025-02-011610.3389/fphar.2025.15302681530268Novel human single-domain antibodies exert potent anti-tumor activity by targeting EGF-like repeat epitope of EpCAMXiaofeng Zhou0Zhifang Liu1Weixiong Zhang2Lin Dai3Tao Chen4Zexiong Lin5Hong Pan6Qi Qi7Henry Wei8State Key Laboratory of Bioactive Molecules and Druggability Assessment, MOE Key Laboratory of Tumor Molecular Biology, Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, Department of Pharmacology, School of Medicine, Jinan University, Guangzhou, ChinaDepartment of Cell Biology and Institute of Biomedicine, National Engineering Research Center of Genetic Medicine, Guangdong Provincial Key Laboratory of Bioengineering Medicine, College of Life Science and Technology, Jinan University, Guangzhou, Guangdong, ChinaDepartment of Cell Biology and Institute of Biomedicine, National Engineering Research Center of Genetic Medicine, Guangdong Provincial Key Laboratory of Bioengineering Medicine, College of Life Science and Technology, Jinan University, Guangzhou, Guangdong, ChinaDepartment of Cell Biology and Institute of Biomedicine, National Engineering Research Center of Genetic Medicine, Guangdong Provincial Key Laboratory of Bioengineering Medicine, College of Life Science and Technology, Jinan University, Guangzhou, Guangdong, ChinaDepartment of Cell Biology and Institute of Biomedicine, National Engineering Research Center of Genetic Medicine, Guangdong Provincial Key Laboratory of Bioengineering Medicine, College of Life Science and Technology, Jinan University, Guangzhou, Guangdong, ChinaDepartment of Cell Biology and Institute of Biomedicine, National Engineering Research Center of Genetic Medicine, Guangdong Provincial Key Laboratory of Bioengineering Medicine, College of Life Science and Technology, Jinan University, Guangzhou, Guangdong, ChinaDepartment of Ophthalmology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, ChinaState Key Laboratory of Bioactive Molecules and Druggability Assessment, MOE Key Laboratory of Tumor Molecular Biology, Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, Department of Pharmacology, School of Medicine, Jinan University, Guangzhou, ChinaDepartment of Cell Biology and Institute of Biomedicine, National Engineering Research Center of Genetic Medicine, Guangdong Provincial Key Laboratory of Bioengineering Medicine, College of Life Science and Technology, Jinan University, Guangzhou, Guangdong, ChinaIntroductionEpCAM (Epithelial cell adhesion molecule) is a key cancer stem cell marker involved in cancer progression, making it an important target for both diagnosis and therapy. Despite efforts using anti-EpCAM monoclonal antibodies (mAbs), their anti-tumor effects have been limited. Single-domain antibodies (sdAbs), in contrast, offer advantages such as efficient tumor penetration and reduced immunogenicity. This study aims to screen and explore novel sdAbs targeting EpCAM for cancer therapy.MethodsA critical EGF-like repeat epitope on the EpCAM extracellular domain was selected for screening a human sdAb library via phage display. The selected sdAbs were purified and their anti-cancer activity was validated through specific binding with the EpCAM peptide. The effects of these sdAbs on cell proliferation, migration, invasion, and apoptosis were tested in vitro, and their anti-tumor activity was assessed in a xenograft model.ResultsFive fully human anti-EpCAM sdAbs were isolated, all of which specifically bound to the EpCAM peptide and showed selective binding to various cancer cell lines, but not to 293T and 3T3 cells. Functional assays demonstrated that these sdAbs significantly inhibited cancer cell proliferation, migration, and invasion, and induced apoptosis. Notably, two sdAbs (aEP3D4 and aEP4G2) exhibited potent anti-tumor effects in vivo, significantly reducing tumor volume and weight in a mouse xenograft model.DiscussionThis study provides compelling evidence that targeting EpCAM with sdAbs is a promising approach for cancer treatment. The identified anti-EpCAM sdAbs exhibit substantial anti-tumor activity both in vitro and in vivo, suggesting they are strong candidates for future therapeutic applications in cancer therapy.https://www.frontiersin.org/articles/10.3389/fphar.2025.1530268/fullEpCAMantibodysingle-domain antibodycancer stem cellscancer therapy
spellingShingle Xiaofeng Zhou
Zhifang Liu
Weixiong Zhang
Lin Dai
Tao Chen
Zexiong Lin
Hong Pan
Qi Qi
Henry Wei
Novel human single-domain antibodies exert potent anti-tumor activity by targeting EGF-like repeat epitope of EpCAM
Frontiers in Pharmacology
EpCAM
antibody
single-domain antibody
cancer stem cells
cancer therapy
title Novel human single-domain antibodies exert potent anti-tumor activity by targeting EGF-like repeat epitope of EpCAM
title_full Novel human single-domain antibodies exert potent anti-tumor activity by targeting EGF-like repeat epitope of EpCAM
title_fullStr Novel human single-domain antibodies exert potent anti-tumor activity by targeting EGF-like repeat epitope of EpCAM
title_full_unstemmed Novel human single-domain antibodies exert potent anti-tumor activity by targeting EGF-like repeat epitope of EpCAM
title_short Novel human single-domain antibodies exert potent anti-tumor activity by targeting EGF-like repeat epitope of EpCAM
title_sort novel human single domain antibodies exert potent anti tumor activity by targeting egf like repeat epitope of epcam
topic EpCAM
antibody
single-domain antibody
cancer stem cells
cancer therapy
url https://www.frontiersin.org/articles/10.3389/fphar.2025.1530268/full
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