Protein kinase D drives the secretion of invasion mediators in triple-negative breast cancer cell lines

Alexia Gali; Irene V Bijnsdorp; Sander R Piersma; Thang V Pham; Elena Gutiérrez-Galindo; Fiona Kühnel; Nikos Tsolakos; Connie R Jimenez; Angelika Hausser; Leonidas G Alexopoulos

doi:10.1016/j.isci.2024.108958

Protein kinase D drives the secretion of invasion mediators in triple-negative breast cancer cell lines

iScience. 2024 Jan 17;27(2):108958. doi: 10.1016/j.isci.2024.108958. eCollection 2024 Feb 16.

Authors

Alexia Gali^{1

2}, Irene V Bijnsdorp^{3

4}, Sander R Piersma⁴, Thang V Pham⁴, Elena Gutiérrez-Galindo⁵, Fiona Kühnel⁵, Nikos Tsolakos², Connie R Jimenez⁴, Angelika Hausser^{5

6}, Leonidas G Alexopoulos^{1

2}

Affiliations

¹ Biomedical Systems Laboratory, National Technical University of Athens, 15780 Athens, Greece.
² Protavio Ltd, Demokritos Science Park, 15341 Athens, Greece.
³ Department of Urology, Cancer Center Amsterdam, Cancer Center Amsterdam, Amsterdam UMC, de Boelelaan 1117, Amsterdam 1081 HV, the Netherlands.
⁴ Department of Medical Oncology, Cancer Center Amsterdam, Amsterdam UMC, OncoProteomics Laboratory, de Boelelaan 1117, , Amsterdam 1081 HV, the Netherlands.
⁵ Institute of Cell Biology and Immunology, University of Stuttgart, 70569 Stuttgart, Germany.
⁶ Stuttgart Research Center for Systems Biology, University of Stuttgart, 70569 Stuttgart, Germany.

Abstract

The protein kinase D (PKD) family members regulate the fission of cargo vesicles at the Golgi complex and play a pro-oncogenic role in triple-negative breast cancer (TNBC). Whether PKD facilitates the secretion of tumor-promoting factors in TNBC, however, is still unknown. Using the pharmacological inhibition of PKD activity and siRNA-mediated depletion of PKD2 and PKD3, we identified the PKD-dependent secretome of the TNBC cell lines MDA-MB-231 and MDA-MB-468. Mass spectrometry-based proteomics and antibody-based assays revealed a significant downregulation of extracellular matrix related proteins and pro-invasive factors such as LIF, MMP-1, MMP-13, IL-11, M-CSF and GM-CSF in PKD-perturbed cells. Notably, secretion of these proteins in MDA-MB-231 cells was predominantly controlled by PKD2 and enhanced spheroid invasion. Consistently, PKD-dependent secretion of pro-invasive factors was more pronounced in metastatic TNBC cell lines. Our study thus uncovers a novel role of PKD2 in releasing a pro-invasive secretome.

Keywords: Cancer; Cell biology.