Abstract
Integrins provide an essential bridge between cancer cells and the extracellular matrix, playing a central role in every stage of disease progression. Despite the recognized importance of integrin phosphorylation in several biological processes, the regulatory mechanisms and their relevance remained elusive. Here we engineer a fluorescence resonance energy transfer biosensor for integrin β1 phosphorylation, screening 96 protein tyrosine phosphatases and identifying Shp2 and PTP-PEST as negative regulators to address this gap. Mutation of the integrin NPxY(783/795) sites revealed the importance of integrin phosphorylation for efficient cancer cell invasion, further supported by inhibition of the identified integrin phosphorylation regulators Shp2 and Src kinase. Using proteomics approaches, we uncovered Cofilin as a component of the phosphorylated integrin-Dok1 complex and linked this axis to effective invadopodia formation, a process supporting breast cancer invasion. These data further implicate dynamic modulation of integrin β1 phosphorylation at NPxY sites at different stages of metastatic dissemination.
| Original language | English |
|---|---|
| Pages (from-to) | 1021-1034 |
| Number of pages | 14 |
| Journal | Nature Cell Biology |
| Volume | 27 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - Jun 2025 |
| MoE publication type | A1 Journal article-refereed |
Funding
Open Access funding provided by University of Turku (including Turku University Central Hospital). We thank P. Laasola, J. Siivonen and R. Mahran for technical assistance and the Ivaska lab for scientific discussion and feedback on the manuscript. We also thank N. Pasquier, A. Isomursu and M. Mathieu for providing necessary reagents for the completion of the study, and for critical reading, we thank H. Hamidi. For support with applying the basement membrane invasion assay, we thank R. Staneva and D. Vignjevic (Institut Curie, Paris, France). For services, instrumentation and expertise at Turku Bioscience (University of Turku, Turku, Finland), we thank the Cell Imaging and Cytometry Core, the Turku Proteomics Facility and the Genome Editing Core, all supported by Biocenter Finland. We also thank the Turku Screening Unit lab automation site, part of the DDCB platform supported by Biocenter Finland, for services, instrumentation and expertise. The clone pENTR223-SH3PXD2A was from the ORFeome library at the Genome Biology Unit core facility, supported by HiLIFE and the Faculty of Medicine, University of Helsinki, and Biocenter Finland. The histological methods were performed by the Histology core facility of the Institute of Biomedicine, University of Turku, Finland. This work was supported by the Finnish Cancer Institute (K. Albin Johansson Professorship to J.I.); a Research Council of Finland research project (grant no. 325464 to J.I.) and Centre of Excellence programme (grant nos. 346131 and 364182 to J.I.); the Cancer Foundation Finland (to J.I.); the Sigrid Juselius Foundation (to J.I.); and the Research Council of Finland InFLAMES Flagship Programme (grant nos. 337530 and 357910). J.R.W.C. was supported by the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement (grant no. 841973), a Research Council of Finland postdoctoral research grant (grant no. 338585) and Research Fellowship (grant no. 360775). G.F. was supported by a Research Council of Finland postdoctoral research grant (grant no. 332402) and the Turku Collegium for Science, Medicine and Technology. M.G. was supported by a Worldwide Cancer Research grant (no. 23-0123 to J.I.).