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Targeting the p300/NONO axis sensitizes melanoma cells to BRAF inhibitors

Abstract

BRAF inhibitors (BRAFi) that target BRAF V600E kinase, a driver mutation found in 50% of melanomas, show a significant antitumor response, but the common emergence of acquired resistance remains a challenge. Abnormal expression of RAF isoforms CRAF and ARAF reactivates pERK1/2, which plays crucial roles in the acquisition of resistance of melanoma cells. However, the mechanisms of dysregulation of RAF isoforms in resistant melanoma cells remain unknown. Here, we identified NONO interacted with and stabilized both CRAF and ARAF in melanoma cells, and that NONO was acetylated at 198K by p300 acetyltransferase, which stabilized NONO via antagonizing its ubiquitination/degradation mediated by RNF8. The upregulation of both p300 and NONO promoted the rebound of pERK1/2 and the subsequent resistance of melanoma cells to BRAFi, and the activation of ERK1/2 in turn induced p300 to form a positive feedback loop in resistant melanoma cells. There was a positive correlation between p300 and NONO in resistant melanoma cells and clinical samples, and p300 inhibitor C646 overcame the resistance of resistant melanoma cells to BRAF inhibitors in vitro and in vivo. Our findings reveal that targeting the positive feedback loop of p300-NONO-CRAF/ARAF-pERK1/2 may be excellent strategies to overcome the resistance of BRAF inhibitors for melanoma patients.

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Fig. 1: CRAF and ARAF are up-regulated in resistant melanoma cells and is responsible for the resistance of melanoma cells to BRAFi.
Fig. 2: NONO is up-regulated in resistant melanoma cells and is responsible for the resistance of melanoma cells to BRAFi.
Fig. 3: NONO improves the expression of CRAF and ARAF.
Fig. 4: NONO interacts with CRAF and ARAF.
Fig. 5: NONO promotes the stability of CRAF and ARAF.
Fig. 6: p300 mediated-acetylation modification of NONO at 198K promotes its stability and upregulation in BRAF inhibitor-resistant melanoma cells.
Fig. 7: p300 is up-regulated in resistant melanoma cells and improves the resistance of melanoma cells BRAFi in vitro and in vivo.
Fig. 8: The resistant melanoma creates a positive feedback loop of p300-NONO-CRAF/ARAF-pERK1/2.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (Nos. 81672866, 81960501 and 81560452 to XBL; No. 81760552 to XT); the Natural Science Foundation of Jiangxi Province (No. 20202BAB206041; Nos. 20161BAB205192 and 20171ACB21073 to XBL).

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XBL, GH and TK designed this research; FZ, XT, SF and XL performed experiments; FZ and XT performed coIP, western blotting and the drug treatment experiments; XL and SF performed the in vivo assay, the histopathological analysis and immunohistochemical staining; JS, CJ, YL, RL, RZ, CZ, BY and ZZ established the resistant melanoma cells and assist with data analysis; XBL, TK and GH wrote the manuscript.

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Correspondence to Guofu Huang or Xiao-Bin Lv.

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This study was approved by the institutional ethical review boards of the Third Affiliated Hospital of Nanchang University (ZKY2020025).

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Zhang, F., Tang, X., Fan, S. et al. Targeting the p300/NONO axis sensitizes melanoma cells to BRAF inhibitors. Oncogene 40, 4137–4150 (2021). https://doi.org/10.1038/s41388-021-01834-1

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