Novel kinase regulators of extracellular matrix internalisation identified by high-content screening modulate invasive carcinoma cell migration

• DOI: 10.1371/journal.pbio.3002930
• Publication Date: 2024-12-12T18:38:36Z
• Authors: Montserrat Llanse...; Keqian Nan; Zhe Bao; Rachele Bacchetti; Shengnan Yuan; Joe Tyler; Xavier Le Guezennec; Frederic A. Bard; Elena Rainero
• Keywords: [SDV] Life Sciences [q-bio]; 0303 health sciences; 03 medical and health sciences; QH301-705.5; Methods and Resources; Biology (General)

The interaction between cancer cells and the extracellular matrix (ECM) plays a pivotal role in tumour progression. While degradation of ECM proteins has been well characterised, endocytosis its impact on cell progression, migration, metastasis is poorly understood. internalisation increased invasive breast cells, suggesting it may support invasiveness. However, current high-throughput approaches mainly focus grown plastic 2D, making difficult to apply these study dynamics. Here, we developed high-content screening assay uptake, based use automated coating for generation highly homogeneous pH-sensitive dye image trafficking live cells. We identified that mitogen-activated protein kinase (MAPK) family members, MAP3K1 MAPK11 (p38β), phosphatase 2 (PP2) subunit PPP2R1A were required ECM-bound α2β1 integrin. Mechanistically, show down-regulation sodium/proton exchanger 1 (NHE1), an established macropinocytosis regulator target p38, mediated macropinocytosis. Moreover, disruption α2 integrin, MAP3K1, MAPK11, PPP2R1A, NHE1-mediated significantly impaired migration invasion 2D 3D culture systems. Of note, integrin-bound was targeted lysosomal degradation, which cell-derived matrices. Finally, integrin expression up-regulated pancreatic tumours correlated with poor prognosis patients. Strikingly, higher chemotherapy-resistant Our results integrin/p38 signalling axis as novel endocytosis, drives demonstrating our approach capability identifying regulators invasion.