Fig. 5: CLU-NRG1 fusion gene is targetable by anti ERBB2/ERBB3 pharmacological agents.

a Schematic representation of FMOC02 patient’s CA125 levels, measured after relapse during standard chemotherapy treatments. b Picture of the original FMOC02_1 ascites sample, the tumour spheroids and established FMOC02_1 PDC. Scale bar, 100 µM. c Schematic representation of CLU-NRG1 rearrangement showing genomic structure of the fusion gene and some of the fusion-point spanning sequences. d Waterfall plot showing the most effective approved chemotherapy (grey) and targeted (purple) drugs for FMOC02_1 PDC based on drug sensitivity score (DSS). e Phosphorylation of ERBB receptors and downstream signalling transducers of FMOC02_1 PDC in dose-response experiments. FMOC02_1 PDCs were treated with afatinib or erlotinib (200 nM) for different time points as indicated, followed by cell lysing and immunoblotting with indicated antibodies. β-actin was included as a loading control. f Long-term CFA with FMOC02_1 PDCs cultured for 14 days in the absence or presence of afatinib and erlotinib (20 or 200 nM). Colonies were counted with Image J using the ColonyArea plugin and shown as percentage inhibition by comparison with untreated cells. Shown results are representative of at least two independent experiments. Data are presented as means ± SEMs. p values from one-way ANOVA is shown. g Dose-response curves of afatinib and erlotinib for the three FMOC02 PDCs. h Phosphorylation of ERBB3, ERK and AKT in FMOC02_3 PDCs treated with trastuzumab and pertuzumab for 12 and 24 h (10 μg/ml), followed by protein extraction and immunoblotting. β-tubulin was included as a loading control. i Long-term CFA with FMOC02_3 PDCs cultured for 14 days in the absence or presence of trastuzumab (10 μg/ml), pertuzumab (10 μg/ml) and their combination (both 10 μg/ml). Colonies were counted with Image J using the ColonyArea plugin and shown as percentage inhibition by comparison with untreated. Shown results are representative of at least two independent experiments. Data are presented as means ± SEMs. p values from one-way ANOVA.