Functional Validation of the Putative Oncogenic Activity of <i>PLAU</i>
Federica Sarno,
Désirée Goubert,
Emilie Logie,
Martijn G. S. Rutten,
Mihaly Koncz,
Christophe Deben,
Anita E. Niemarkt,
Lucia Altucci,
Pernette J. Verschure,
Antal Kiss,
Wim Vanden Berghe,
Marianne G. Rots
Affiliations
Federica Sarno
Epigenetic Editing, Department of Pathology and Medical Biology, University Medical Centre Groningen, University of Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands
Désirée Goubert
Epigenetic Editing, Department of Pathology and Medical Biology, University Medical Centre Groningen, University of Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands
Emilie Logie
Center for Oncological Research (CORE), Integrated Personalized & Precision Oncology Network (IPPON), Laboratory of Protein Chemistry, Proteomics and Epigenetic Signalling (PPES), Department of Biomedical Sciences, University of Antwerp, 2610 Wilrijk, Belgium
Martijn G. S. Rutten
Epigenetic Editing, Department of Pathology and Medical Biology, University Medical Centre Groningen, University of Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands
Mihaly Koncz
Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands
Christophe Deben
Center for Oncological Research (CORE), Integrated Personalized & Precision Oncology Network (IPPON), University of Antwerp, 2610 Wilrijk, Belgium
Anita E. Niemarkt
Epigenetic Editing, Department of Pathology and Medical Biology, University Medical Centre Groningen, University of Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands
Lucia Altucci
Department of Precision Medicine, Universita’ degli Studi della Campania “Luigi Vanvitelli”, 80138 Naples, Italy
Pernette J. Verschure
Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands
Antal Kiss
Institute of Biochemistry, Biological Research Centre, H-6726 Szeged, Hungary
Wim Vanden Berghe
Center for Oncological Research (CORE), Integrated Personalized & Precision Oncology Network (IPPON), Laboratory of Protein Chemistry, Proteomics and Epigenetic Signalling (PPES), Department of Biomedical Sciences, University of Antwerp, 2610 Wilrijk, Belgium
Marianne G. Rots
Epigenetic Editing, Department of Pathology and Medical Biology, University Medical Centre Groningen, University of Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands
Plasminogen activator, urokinase (PLAU) is involved in cell migration, proliferation and tissue remodeling. PLAU upregulation is associated with an increase in aggressiveness, metastasis, and invasion of several cancer types, including breast cancer. In patients, this translates into decreased sensitivity to hormonal treatment, and poor prognosis. These clinical findings have led to the examination of PLAU as a biomarker for predicting breast cancer prognosis and therapy responses. In this study, we investigated the functional ability of PLAU to act as an oncogene in breast cancers by modulating its expression using CRISPR-deactivated Cas9 (CRISPR-dCas9) tools. Different effector domains (e.g., transcription modulators (VP64, KRAB)) alone or in combination with epigenetic writers (DNMT3A/3L, MSssI) were fused to dCas9 and targeted to the PLAU promoter. In MDA-MB-231 cells characterized by high PLAU expression downregulation of PLAU expression by CRISPR-dCas9-DNMT3A/3L-KRAB, resulted in decreased cell proliferation. Conversely, CRISPR-dCas9-VP64 induced PLAU upregulation in low PLAU expressing MCF-7 cells and significantly increased aggressiveness and invasion. In conclusion, modulation of PLAU expression affected metastatic related properties of breast cancer cells, thus further validating its oncogenic activity in breast cancer cells.
