Reciprocal priming between receptor tyrosine kinases at recycling endosomes orchestrates cellular signalling outputs

Integration of signalling downstream of individual receptor tyrosine kinases (RTKs) is crucial to fine tune cellular homeostasis during development and in pathological conditions, including breast cancer. However, how signalling integration is regulated and whether the endocytic fate of single receptors controls such signalling integration remains poorly elucidated. Combining quantitative phosphoproteomics and targeted assays, we generated a detailed picture of recycling-dependent fibroblast growth factor (FGF) signalling in breast cancer cells, with a focus on distinct FGF receptors (FGFRs). We discovered reciprocal priming between FGFRs and epidermal growth factor (EGF) receptor (EGFR) that is coordinated at recycling endosomes. FGFR recycling ligands induce EGFR phosphorylation on threonine 693. This phosphorylation event alters both FGFR and EGFR trafficking and primes FGFR-mediated proliferation but not cell invasion. In turn, FGFR signalling primes EGF-mediated outputs via EGFR threonine 693 phosphorylation. This reciprocal priming between distinct families of RTKs from recycling endosomes exemplifies a novel signalling integration hub where recycling endosomes orchestrate cellular behaviour. Therefore, targeting reciprocal priming over individual receptors may improve personalized therapies in breast and other cancers.

[1]  R. Aebersold,et al.  Proteomic and interactomic insights into the molecular basis of cell functional diversity , 2020, Nature Reviews Molecular Cell Biology.

[2]  D. Generali,et al.  The Fibroblast Growth Factor Receptors in Breast Cancer: from Oncogenesis to Better Treatments , 2020, International journal of molecular sciences.

[3]  Seung-Oe Lim,et al.  Oncogenic signaling pathways associated with immune evasion and the resistance to immune checkpoint inhibitors in cancer. , 2019, Seminars in cancer biology.

[4]  I. Meinhold-Heerlein,et al.  Molecular Targeting Therapy against EGFR Family in Breast Cancer: Progress and Future Potentials , 2019, Cancers.

[5]  L. Mulligan,et al.  GGA3-mediated recycling of the RET receptor tyrosine kinase contributes to cell migration and invasion , 2019, Oncogene.

[6]  Jan C. Refsgaard,et al.  Oncogenic Mutations Rewire Signaling Pathways by Switching Protein Recruitment to Phosphotyrosine Sites , 2019, Cell.

[7]  M. Gnant,et al.  Breast cancer , 2019, Nature Reviews Disease Primers.

[8]  Jing Wang,et al.  WebGestalt 2019: gene set analysis toolkit with revamped UIs and APIs , 2019, Nucleic Acids Res..

[9]  Florian Gnad,et al.  The Case for Proteomics and Phospho‐Proteomics in Personalized Cancer Medicine , 2019, Proteomics. Clinical applications.

[10]  S. Munro,et al.  In vivo identification of GTPase interactors by mitochondrial relocalization and proximity biotinylation , 2019, bioRxiv.

[11]  M. Mohammadi,et al.  Structural Biology of the FGF7 Subfamily , 2019, Front. Genet..

[12]  M. Yaffe.  Why geneticists stole cancer research even though cancer is primarily a signaling disease , 2019, Science Signaling.

[13]  J. Otlewski,et al.  Targeting Cellular Trafficking of Fibroblast Growth Factor Receptors as a Strategy for Selective Cancer Treatment , 2018, Journal of clinical medicine.

[14]  Damian Szklarczyk,et al.  STRING v11: protein–protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets , 2018, Nucleic Acids Res..

[15]  Martin Eisenacher,et al.  The PRIDE database and related tools and resources in 2019: improving support for quantification data , 2018, Nucleic Acids Res..

[16]  R. Grose,et al.  Emerging Roles of Fibroblast Growth Factor 10 in Cancer , 2018, Front. Genet..

[17]  C. Francavilla,et al.  Regulation of FGF10 Signaling in Development and Disease , 2018, Front. Genet..

[18]  Lucas Pelkmans,et al.  Multiplexed protein maps link subcellular organization to cellular states , 2018, Science.

[19]  P. Bastiaens,et al.  Contact inhibitory Eph signaling suppresses EGF-promoted cell migration by decoupling EGFR activity from vesicular recycling , 2018, Science Signaling.

[20]  Martin P. Seybold,et al.  Complex polymorphisms in endocytosis genes suggest alpha-cyclodextrin as a treatment for breast cancer , 2018, PloS one.

[21]  Anne E Carpenter,et al.  CellProfiler 3.0: Next-generation image processing for biology , 2018, PLoS biology.

[22]  Nandini A. Sahasrabuddhe,et al.  Quantitative phosphoproteomic analysis reveals reciprocal activation of receptor tyrosine kinases between cancer epithelial cells and stromal fibroblasts , 2018, Clinical Proteomics.

[23]  Dhiraj Kumar,et al.  Receptor tyrosine kinases (RTKs) in breast cancer: signaling, therapeutic implications and challenges , 2018, Molecular Cancer.

[24]  C. Lovly,et al.  Mechanisms of receptor tyrosine kinase activation in cancer , 2018, Molecular Cancer.

[25]  Hans Clevers,et al.  A Living Biobank of Breast Cancer Organoids Captures Disease Heterogeneity , 2018, Cell.

[26]  Ying Cheng,et al.  Current Development Status of MEK Inhibitors , 2017, Molecules.

[27]  S. Schmid.  Reciprocal regulation of signaling and endocytosis: Implications for the evolving cancer cell , 2017, The Journal of cell biology.

[28]  M. Landthaler,et al.  An immediate–late gene expression module decodes ERK signal duration , 2017, Molecular systems biology.

[29]  P. P. Di Fiore,et al.  Behind the Scenes: Endo/Exocytosis in the Acquisition of Metastatic Traits. , 2017, Cancer research.

[30]  Nevan J. Krogan,et al.  An Approach to Spatiotemporally Resolve Protein Interaction Networks in Living Cells , 2017, Cell.

[31]  C. Sotiriou,et al.  HER2-Overexpressing Breast Cancers Amplify FGFR Signaling upon Acquisition of Resistance to Dual Therapeutic Blockade of HER2 , 2017, Clinical Cancer Research.

[32]  N. Turner,et al.  Advances and challenges in targeting FGFR signalling in cancer , 2017, Nature Reviews Cancer.

[33]  Li Ding,et al.  Proteogenomic integration reveals therapeutic targets in breast cancer xenografts , 2017, Nature Communications.

[34]  A. Wardley,et al.  Patient-derived Mammosphere and Xenograft Tumour Initiation Correlates with Progression to Metastasis , 2016, Journal of Mammary Gland Biology and Neoplasia.

[35]  Sophie Dahan,et al.  Spatial and Temporal Regulation of Receptor Tyrosine Kinase Activation and Intracellular Signal Transduction. , 2016, Annual review of biochemistry.

[36]  Kristoffer T G Rigbolt,et al.  Multilayered proteomics reveals molecular switches dictating ligand-dependent EGFR trafficking , 2016, Nature Structural &Molecular Biology.

[37]  Michael L. Gatza,et al.  Proteogenomics connects somatic mutations to signaling in breast cancer , 2016, Nature.

[38]  Boris N. Kholodenko,et al.  Feedback regulation in cell signalling: Lessons for cancer therapeutics. , 2016, Seminars in cell & developmental biology.

[39]  Xuetao Cao,et al.  Self-regulation and cross-regulation of pattern-recognition receptor signalling in health and disease , 2015, Nature Reviews Immunology.

[40]  P. Bastiaens,et al.  EGF-dependent re-routing of vesicular recycling switches spontaneous phosphorylation suppression to EGFR signaling , 2015, eLife.

[41]  Tammy J. Toney-Butler.  Physiology, Tyrosine Kinase Receptors , 2015 .

[42]  D. Hochhauser,et al.  WASH and Tsg101/ALIX-dependent diversion of stress-internalized EGFR from the canonical endocytic pathway , 2015, Nature Communications.

[43]  Z. Fayad,et al.  The effect of BMS-582949, a P38 mitogen-activated protein kinase (P38 MAPK) inhibitor on arterial inflammation: a multicenter FDG-PET trial. , 2015, Atherosclerosis.

[44]  N. Itoh,et al.  The Fibroblast Growth Factor signaling pathway , 2015, Wiley interdisciplinary reviews. Developmental biology.

[45]  Janos X. Binder,et al.  DISEASES: Text mining and data integration of disease–gene associations , 2014, bioRxiv.

[46]  Marco Y. Hein,et al.  Accurate Proteome-wide Label-free Quantification by Delayed Normalization and Maximal Peptide Ratio Extraction, Termed MaxLFQ * , 2014, Molecular & Cellular Proteomics.

[47]  Lucas Pelkmans,et al.  A Hierarchical Map of Regulatory Genetic Interactions in Membrane Trafficking , 2014, Cell.

[48]  S. Kermorgant,et al.  Met endosomal signalling: in the right place, at the right time. , 2014, The international journal of biochemistry & cell biology.

[49]  Blagoy Blagoev,et al.  Functional proteomics defines the molecular switch underlying FGF receptor trafficking and cellular outputs. , 2013, Molecular cell.

[50]  A. Sorkin,et al.  Endocytosis of receptor tyrosine kinases. , 2013, Cold Spring Harbor perspectives in biology.

[51]  J. Otlewski,et al.  ERK-Mediated Phosphorylation of Fibroblast Growth Factor Receptor 1 on Ser777 Inhibits Signaling , 2013, Science Signaling.

[52]  S. Wiemann,et al.  Combinatorial targeting of FGF and ErbB receptors blocks growth and metastatic spread of breast cancer models , 2013, Breast Cancer Research.

[53]  A. Cardona,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[54]  Paul H. Huang.  Phosphoproteomic studies of receptor tyrosine kinases: future perspectives. , 2012, Molecular bioSystems.

[55]  K. Coulonval,et al.  Coupling of T161 and T14 phosphorylations protects cyclin B–CDK1 from premature activation , 2011, Molecular biology of the cell.

[56]  J. Taminau,et al.  InSilico DB: an online platform to collaboratively structure and export publicly available datasets from the Gene Expression Omnibus database , 2011, Genome Biology.

[57]  M. Mann,et al.  Andromeda: a peptide search engine integrated into the MaxQuant environment. , 2011, Journal of proteome research.

[58]  Y. Kalaidzidis,et al.  Systems survey of endocytosis by multiparametric image analysis , 2010, Nature.

[59]  Ario de Marco,et al.  The binding of NCAM to FGFR1 induces a specific cellular response mediated by receptor trafficking , 2009, The Journal of cell biology.

[60]  Eric O. Aboagye,et al.  The fibroblast growth factor receptor inhibitor PD173074 blocks small cell lung cancer growth in vitro and in vivo. , 2009, Cancer research.

[61]  M. Mann,et al.  MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification , 2008, Nature Biotechnology.

[62]  J. Norman,et al.  Endocytic transport of integrins during cell migration and invasion. , 2008, Trends in cell biology.

[63]  Hiroaki Sakurai,et al.  Transient Suppression of Ligand-mediated Activation of Epidermal Growth Factor Receptor by Tumor Necrosis Factor-α through the TAK1-p38 Signaling Pathway* , 2007, Journal of Biological Chemistry.

[64]  Wen-Lin Kuo,et al.  A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. , 2006, Cancer cell.

[65]  M. Mann,et al.  Global, In Vivo, and Site-Specific Phosphorylation Dynamics in Signaling Networks , 2006, Cell.

[66]  Y. Yarden,et al.  p38 MAP kinase mediates stress‐induced internalization of EGFR: implications for cancer chemotherapy , 2006, The EMBO journal.

[67]  A. Sharrocks.  Cell Cycle: Sustained ERK Signalling Represses the Inhibitors , 2006, Current Biology.

[68]  A. Brech,et al.  Different intracellular trafficking of FGF1 endocytosed by the four homologous FGF receptors , 2005, Journal of Cell Science.

[69]  C. Plass,et al.  Variability in organ-specific EGFR mutational spectra in tumour epithelium and stroma may be the biological basis for differential responses to tyrosine kinase inhibitors , 2005, British Journal of Cancer.

[70]  Sylvain V Costes,et al.  Automatic and quantitative measurement of protein-protein colocalization in live cells. , 2004, Biophysical journal.

[71]  Salvatore Pece,et al.  Endocytosis and cancer. , 2004, Current opinion in cell biology.

[72]  P. Shannon,et al.  Cytoscape: a software environment for integrated models of biomolecular interaction networks. , 2003, Genome research.

[73]  Peg Coughlin,et al.  Identification of XMAP215 as a microtubule-destabilizing factor in Xenopus egg extract by biochemical purification , 2003, The Journal of cell biology.

[74]  Terence P. Speed,et al.  A comparison of normalization methods for high density oligonucleotide array data based on variance and bias , 2003, Bioinform..

[75]  J. Schlessinger.  Cell Signaling by Receptor Tyrosine Kinases , 2000, Cell.

[76]  W. Cavenee,et al.  Tyrphostin AG 1478 preferentially inhibits human glioma cells expressing truncated rather than wild-type epidermal growth factor receptors. , 1996, Cancer research.

[77]  H. Wiley,et al.  Mutational removal of the Thr669 and Ser671 phosphorylation sites alters substrate specificity and ligand-induced internalization of the epidermal growth factor receptor. , 1990, The Journal of biological chemistry.

[78]  PhD Ernest Heimsath,et al.  Cell Migration and Invasion , 2022 .

[79]  I. Dragoni,et al.  PAK 4 regulates stemness and progression in endocrine resistant ER-positive metastatic breast cancer , 2019 .

[80]  Stefka Tyanova,et al.  Perseus: A Bioinformatics Platform for Integrative Analysis of Proteomics Data in Cancer Research. , 2018, Methods in molecular biology.

[81]  M. Miączyńska,et al.  Integration of the Endocytic System into the Network of Cellular Functions. , 2018, Progress in molecular and subcellular biology.

[82]  A. Tan,et al.  Exploiting receptor tyrosine kinase co-activation for cancer therapy. , 2017, Drug discovery today.

[83]  C. Sotiriou,et al.  HER 2-Overexpressing Breast Cancers Amplify FGFR Signaling upon Acquisition of Resistance to Dual Therapeutic Blockade of HER 2 , 2017 .

[84]  J. Peterson,et al.  Cell and molecular biology of epidermal growth factor receptor. , 2014, International review of cell and molecular biology.

[85]  B. Spencer‐Dene,et al.  The Fibroblast Growth Factor Receptor Inhibitor PD 173074 Blocks Small Cell Lung Cancer Growth In vitro and In vivo , 2009 .

[86]  S. Winograd-Katz,et al.  Cisplatin induces PKB/Akt activation and p38MAPK phosphorylation of the EGF receptor , 2007, Oncogene.

[87]  J Strackee,et al.  Largest contour segmentation: a tool for the localization of spots in confocal images. , 1996, Cytometry.

[88]  W. Fantl,et al.  Signalling by receptor tyrosine kinases. , 1993, Annual review of biochemistry.