Development and validation of a high-throughput screening pipeline of compound libraries to target EMT

Sven Jonckheere^{1

2

3}, Joachim Taminau^{1

2

3}, Jamie Adams⁴, Jef Haerinck^{1

2

3}, Jordy De Coninck^{1

2

3}, Jeroen Verstappe^{1

2

3}, Kato De Clercq^{1

2

3}, Evelien Peeters^{1

2

3

5}, Alexander Gheldof^{2

3

6}, Eva De Smedt^{1

2

3}, Vera Goossens⁷, Dominique Audenaert⁷, Aurélie Candi⁸, Matthias Versele⁸, Dominic De Groote¹, Hanne Verschuere^{1

2

3

9}, Marc Stemmler¹⁰, Thomas Brabletz¹⁰, Peter Vandenabeele^{1

2

9}, Andreu Casali¹¹, Kyra Campbell⁴, Steven Goossens^#^{1

5}, Geert Berx^#^{12

13

14}

Affiliations

¹ Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium.
² Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
³ Molecular and Cellular Oncology Laboratory, Ghent University, Ghent, Belgium.
⁴ Department of Biomedical Science, The University of Sheffield, Sheffield, UK.
⁵ Department of Diagnostic Sciences, Ghent University, Ghent, Belgium.
⁶ Centre for Medical Genetics, Vrije Universiteit Brussel (VUB), Universitair Ziekenhuis Brussel (UZ Brussel), Brussels, Belgium.
⁷ VIB Screening Core & UGent Expertise Centre for Bioassay Development and Screening (C-BIOS), Ghent University, Ghent, Belgium.
⁸ Cistim Leuven vzw & Centre for Drug Design and Discovery (CD3), KU Leuven, Leuven, Belgium.
⁹ Cell Death and Inflammation Unit, VIB Center for Inflammation Research, Ghent, Belgium.
¹⁰ Department of Experimental Medicine 1, Nikolaus-Fiebiger Center for Molecular Medicine, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany.
¹¹ Department de Ciènces Mèdiques Bàsiques, Universitat de Llieda and IRBLleida, Lleida, Spain.
¹² Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium. Geert.Berx@ugent.be.
¹³ Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium. Geert.Berx@ugent.be.
¹⁴ Molecular and Cellular Oncology Laboratory, Ghent University, Ghent, Belgium. Geert.Berx@ugent.be.

^# Contributed equally.

PMID: 40341726
DOI: 10.1038/s41418-025-01515-6

Development and validation of a high-throughput screening pipeline of compound libraries to target EMT

Sven Jonckheere et al. Cell Death Differ. 2025.

. 2025 May 8.

doi: 10.1038/s41418-025-01515-6. Online ahead of print.

Authors

Affiliations

¹ Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium.
² Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
³ Molecular and Cellular Oncology Laboratory, Ghent University, Ghent, Belgium.
⁴ Department of Biomedical Science, The University of Sheffield, Sheffield, UK.
⁵ Department of Diagnostic Sciences, Ghent University, Ghent, Belgium.
⁶ Centre for Medical Genetics, Vrije Universiteit Brussel (VUB), Universitair Ziekenhuis Brussel (UZ Brussel), Brussels, Belgium.
⁷ VIB Screening Core & UGent Expertise Centre for Bioassay Development and Screening (C-BIOS), Ghent University, Ghent, Belgium.
⁸ Cistim Leuven vzw & Centre for Drug Design and Discovery (CD3), KU Leuven, Leuven, Belgium.
⁹ Cell Death and Inflammation Unit, VIB Center for Inflammation Research, Ghent, Belgium.
¹⁰ Department of Experimental Medicine 1, Nikolaus-Fiebiger Center for Molecular Medicine, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany.
¹¹ Department de Ciènces Mèdiques Bàsiques, Universitat de Llieda and IRBLleida, Lleida, Spain.
¹² Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium. Geert.Berx@ugent.be.
¹³ Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium. Geert.Berx@ugent.be.
¹⁴ Molecular and Cellular Oncology Laboratory, Ghent University, Ghent, Belgium. Geert.Berx@ugent.be.

^# Contributed equally.

PMID: 40341726
DOI: 10.1038/s41418-025-01515-6

Abstract

Epithelial to Mesenchymal transitions (EMT) drive cell plasticity and are associated with cell features such as invasiveness, migration and stemness. They are orchestrated by select families of EMT-associated transcription factors, which exhibit pleiotropic roles in the malignant progression of various cancer types, such as breast and colorectal cancer (CRC). This has spurred interest in EMT as a promising target for the development of novel therapeutic strategies. In this study, we developed a phenotypic dual EMT Sensor screening assay, amendable to efficient high-throughput identification of small molecules interfering with EMT. In a proof-of-concept screening we identified anti-EMT repurposing drugs. From these, we validated RepSox, a selective inhibitor of the TGF-β type I receptor ALK5, and demonstrated that it is potently blocking EMT in both breast and colorectal cancer cell lines in vitro. In addition, utilizing a Drosophila melanogaster metastatic CRC model we confirmed the ability of the identified anti-EMT hits to suppress metastatic behavior in vivo.

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Conflict of interest statement

Competing interests: The authors declare no competing interests. Ethics approval: All methods were performed in accordance with the relevant guidelines and regulations. The manuscript reports studies that did not involve human participants, human data, or human tissue. All animal experiments were conducted in compliance with ethical standards. Mice were housed according to institutional guidelines for the care and use of laboratory animals. All breeding and experimental procedures were approved by the local ethics committee (EC) before the start of the experiment under the following application numbers: EC2018-027 and EC2019-069. This research involved the use of the invertebrate insect Drosophila melanogaster in the UK. According to the UK’s Animals (Scientific Procedures) Act 1986, as amended in 2012, Drosophila melanogaster is not classified as a ‘protected animal.’

References

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1. Yang J, Antin P, Berx G, Blanpain C, Brabletz T, Bronner M, et al. Guidelines and definitions for research on epithelial-mesenchymal transition. Nat Rev Mol Cell Biol. 2020;21:341–52. - PubMed - PMC - DOI
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Development and validation of a high-throughput screening pipeline of compound libraries to target EMT

Affiliations

Development and validation of a high-throughput screening pipeline of compound libraries to target EMT

Authors

Affiliations

Abstract

Conflict of interest statement

References

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