Spatially resolved phosphoproteomics reveals fibroblast growth factor receptor recycling-driven regulation of autophagy and survival

• Published in: Nature communications 2022-11, Vol.13 (1), p.6589-6589, Article 6589
• Main Authors: Watson, Joanne, Ferguson, Harriet R., Brady, Rosie M., Ferguson, Jennifer, Fullwood, Paul, Mo, Hanyi, Bexley, Katherine H., Knight, David, Howell, Gareth, Schwartz, Jean-Marc, Smith, Michael P., Francavilla, Chiara
• Format: Article
• Language: English
• Subjects: 13; 13/106; 13/31; 14/19; 631/337/458/1733; 631/45/475; 631/80/313/1461; 631/80/39; 82/58; 96/95; Autophagy; Bioinformatics; Biotinylation; Cancer; Cell fate; Cell proliferation; Cell survival; Endocytosis; Endocytosis - physiology; Endosomes; Endosomes - metabolism; Fibroblast growth factor 10; Fibroblast Growth Factor 10 - metabolism; Fibroblast growth factor receptors; Fibroblasts; Growth factors; Homeostasis; Humanities and Social Sciences; Kinases; Ligands; Membranes; multidisciplinary; Peptides; Protein turnover; Protein-tyrosine kinase receptors; Receptor, Fibroblast Growth Factor, Type 2 - metabolism; Receptors; Recruitment; Science; Science (multidisciplinary); Signaling; Survival; TOR protein; Tyrosine
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-022-34298-2

Receptor Tyrosine Kinase (RTK) endocytosis-dependent signalling drives cell proliferation and motility during development and adult homeostasis, but is dysregulated in diseases, including cancer. The recruitment of RTK signalling partners during endocytosis, specifically during recycling to the plasma membrane, is still unknown. Focusing on Fibroblast Growth Factor Receptor 2b (FGFR2b) recycling, we reveal FGFR signalling partners proximal to recycling endosomes by developing a Spatially Resolved Phosphoproteomics (SRP) approach based on APEX2-driven biotinylation followed by phosphorylated peptides enrichment. Combining this with traditional phosphoproteomics, bioinformatics, and targeted assays, we uncover that FGFR2b stimulated by its recycling ligand FGF10 activates mTOR-dependent signalling and ULK1 at the recycling endosomes, leading to autophagy suppression and cell survival. This adds to the growing importance of RTK recycling in orchestrating cell fate and suggests a therapeutically targetable vulnerability in ligand-responsive cancer cells. Integrating SRP with other systems biology approaches provides a powerful tool to spatially resolve cellular signalling. Recruitment of Receptor Tyrosine Kinase signalling partners during endocytosis, specifically during recycling to the plasma membrane, is crucial to signal propagation and regulation. Here, the authors reveal FGFR signalling partners proximal to recycling endosomes with a spatially resolved phosphoproteomics approach.