SB202190 Predicts BRAF-Activating Mutations in Primary Colorectal Cancer Organoids via Erk1-2 Modulation

• DOI: 10.3390/cells12040664
• Publication Date: 2023-02-20T10:27:01Z
• Authors: Delfina Costa; Roberta Venè; Simona Coco; Luca Longo; Francesca Tosetti; Stefano Scabini; Luca Mastracci; Federica Grillo; Alessandro Poggi; Roberto Benelli
• Keywords: Proto-Oncogene Proteins B-raf; 0301 basic medicine; QH573-671; organoid; colorectal cancer; Erk1-2; Article; BRAF; 3. Good health; SB202190; ErbB Receptors; Organoids; 03 medical and health sciences; BRAF; colorectal cancer; Erk1-2; organoid; SB202190; Mutation; Humans; Cytology; Colorectal Neoplasms; Protein Kinase Inhibitors

The p38 inhibitor SB202190 is a necessary component of the medium used for normal colorectal mucosa cultures. Sato et al. suggested that the primary activity of SB202190 may be EGFR signaling stabilization, causing an increased phosphorylation of Erk1-2 sustaining organoid proliferation. However, the growth of some colorectal cancer (CRC)-derived organoid cultures is inhibited by this molecule via an unknown mechanism. We biochemically investigated SB202190 activity on a collection of 25 primary human CRC organoids, evaluating EGFR, Akt and Erk1-2 activation using Western blot. We found that Erk1-2 phosphorylation was induced by SB202190 in 20 organoid cultures and inhibited in 5 organoid cultures. A next-generation sequencing (NGS) analysis revealed that the inhibition of p-Erk1-2 signaling corresponded to the cultures with BRAF mutations (with four different hits, one being undescribed), while p-Erk1-2 induction was apparently unrelated to other mutations involving the EGFR pathway (Her2, KRAS and NRAS). We found that SB202190 mirrored the biochemical activity of the BRAF inhibitor Dabrafenib, known to induce the paradoxical activation of p-Erk1-2 signaling in BRAF wild-type cells. SB202190 was a more effective inhibitor of BRAF-mutated organoid growth in the long term than the specific BRAF inhibitors Dabrafenib and PLX8394. Overall, SB202190 can predict BRAF-activating mutations in patient-derived organoids, as well as allowing for the identification of new BRAF variants, preceding and enforcing NGS data.