Journal Article

Quantifying single-cell ERK dynamics in colorectal cancer organoids reveals EGFR as an amplifier of oncogenic MAPK pathway signalling

Sassi, Francesco, Vries, Rob G. J., van Ineveld, Ravian L., Rehmann, Holger, Kersten, Simone, Cappe, Benjamin, Bos, Johannes L., Sipieter, François, Snippert, Hugo J. G., Mertens, Sander, Laskaris, Dimitrios, Post, Jasmin B., Boj, Sylvia F., Ponsioen, Bas, Bertotti, Andrea, Vandenabeele, Peter, Riquet, Franck B., Trusolino, Livio, Buissant des Amorie, Julian R., Verlaan-Klink, Ingrid

2021

Overview

Direct targeting of the downstream mitogen-activated protein kinase (MAPK) pathway to suppress extracellular-regulated kinase (ERK) activation in KRAS and BRAF mutant colorectal cancer (CRC) has proven clinically unsuccessful, but promising results have been obtained with combination therapies including epidermal growth factor receptor (EGFR) inhibition. To elucidate the interplay between EGF signalling and ERK activation in tumours, we used patient-derived organoids (PDOs) from KRAS and BRAF mutant CRCs. PDOs resemble in vivo tumours, model treatment response and are compatible with live-cell microscopy. We established real-time, quantitative drug response assessment in PDOs with single-cell resolution, using our improved fluorescence resonance energy transfer (FRET)-based ERK biosensor EKAREN5. We show that oncogene-driven signalling is strikingly limited without EGFR activity and insufficient to sustain full proliferative potential. In PDOs and in vivo, upstream EGFR activity rigorously amplifies signal transduction efficiency in KRAS or BRAF mutant MAPK pathways. Our data provide a mechanistic understanding of the effectivity of EGFR inhibitors within combination therapies against KRAS and BRAF mutant CRC. Ponsioen et al. use a FRET‐based ERK biosensor EKAREN5 in patient‐derived organoids to show that EGFR activity amplifies signal transduction efficiency in KRAS or BRAF mutant MAPK pathways.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 13/100 / 14/33 / 14/63 / 631/80/86/2368 / 692/699/67/1504/1885 / Biomedical and Life Sciences / Biosensors / Cancer / Cancer Research / Cell Biology / Cell Line, Tumor / Cellular signal transduction / Colorectal cancer / Colorectal Neoplasms - drug therapy / Colorectal Neoplasms - genetics / Colorectal Neoplasms - pathology / Development and progression / Developmental Biology / Energy transfer / Epidermal growth factor / Epidermal growth factor receptors / ErbB Receptors - genetics / Extracellular signal-regulated kinase / Fluorescence / Fluorescence resonance energy transfer / Gene Expression Regulation, Neoplastic - drug effects / Genetic aspects / Growth factors / Health aspects / Humans / K-Ras protein / Kinases / Life Sciences / MAP kinase / MAP Kinase Signaling System - drug effects / Mitogen-Activated Protein Kinase Kinases - genetics / Mitogen-activated protein kinases / Mutants / Mutation / Oncogenes / Oncology, Experimental / Organoids / Organoids - metabolism / Organoids - pathology / Organs, Culture of / Protein kinase / Protein Kinase Inhibitors - pharmacology / Proto-Oncogene Proteins B-raf - genetics / Proto-Oncogene Proteins p21(ras) - genetics / Signal transduction / Signaling / Single-Cell Analysis / Stem Cells / Tumors