In cancer, the activating transcription factor 2 (ATF2) has pleiotropic functions in cellular responses to growth stimuli, damage, or inflammation. Due to only limited studies, the significance of ATF2 in colorectal cancer (CRC) is not well understood. We report that low ATF2 levels correlated with worse prognosis and tumor aggressiveness in CRC patients. NanoString gene expression and ChIP analysis confirmed trophoblast cell surface antigen 2 (TROP2) as a novel inhibitory ATF2 target gene. This inverse correlation was further observed in primary human tumor tissues. Immunostainings revealed that high intratumoral heterogeneity for ATF2 and TROP2 expression was sustained also in liver metastasis. Mechanistically, our in vitro data of CRISPR/Cas9-generated ATF2 knockout (KO) clones revealed that high TROP2 levels were critical for cell de-adhesion and increased cell migration without triggering EMT. TROP2 was enriched in filopodia and displaced Paxillin from adherens junctions. In vivo imaging, micro-computer tomography, and immunostainings verified that an ATF2KO/TROP2high status triggered tumor invasiveness in in vivo mouse and chicken xenograft models. In silico analysis provided direct support that ATF2low/TROP2high expression status defined high-risk CRC patients. Finally, our data demonstrate that ATF2 acts as a tumor suppressor by inhibiting the cancer driver TROP2. Therapeutic TROP2 targeting might prevent particularly the first steps in metastasis, i.e., the de-adhesion and invasion of colon cancer cells.

Comprehensive Cancer Center Erlangen EMN University Hospital Erlangen Friedrich Alexander University Erlangen Nürnberg 91054 Erlangen Germany

Czech Center for Phenogenomics Institute of Molecular Genetics of the ASCR v v i 142 20 Prague Czech Republic

Department of Biological Sciences Middle East Technical University 06800 Ankara Turkey

Department of Bioscience and Bioengineering Indian Institute of Technology Jodhpur Jodhpur 342037 India

Department of Chemistry and Pharmacy Molecular and Clinical Pharmacy Friedrich Alexander University Erlangen Nürnberg 91058 Erlangen Germany

Department of Nephropathology Institute of Pathology University Hospital Erlangen Friedrich Alexander University Erlangen Nürnberg 91054 Erlangen Germany

Department of Pathology and Legal Medicine Federal University of Health Sciences of Porto Alegre Porto Alegre 90050 170 Brazil

Department of Surgery University Hospital Erlangen Friedrich Alexander University Erlangen Nürnberg 91054 Erlangen Germany

Experimental Tumor Pathology Institute of Pathology University Hospital Erlangen Friedrich Alexander University Erlangen Nürnberg Universitätsstraße 22 91054 Erlangen Germany

Institute of Bioinformatics and Applied Biotechnology Bangalore 560100 India

Institute of Pathology University Hospital Erlangen Friedrich Alexander University Erlangen Nürnberg 91054 Erlangen Germany

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