Activating KRAS, NRAS, and BRAF mutants enhance proteasome capacity and reduce endoplasmic reticulum stress in multiple myeloma.
KRAS, NRAS, and BRAF mutations which activate p44/42 mitogen-activated protein kinase (MAPK) signaling are found in half of myeloma patients and contribute to proteasome inhibitor (PI) resistance, but the underlying mechanisms are not fully understood. We established myeloma cell lines expressing wild-type (WT), constitutively active (CA) (G12V/G13D/Q61H), or dominant-negative (DN) (S17N)-KRAS and -NRAS, or BRAF-V600E. Cells expressing CA mutants showed increased proteasome maturation protein (POMP) and nuclear factor (erythroid-derived 2)-like 2 (NRF2) expression. This correlated with an increase in catalytically active proteasome subunit β (PSMB)-8, PSMB9, and PSMB10, which occurred in an ETS transcription factor-dependent manner. Proteasome chymotrypsin-like, trypsin-like, and caspase-like activities were increased, and this enhanced capacity reduced PI sensitivity, while DN-KRAS and DN-NRAS did the opposite. Pharmacologic RAF or MAPK kinase (MEK) inhibitors decreased proteasome activity, and sensitized myeloma cells to PIs. CA-KRAS, CA-NRAS, and CA-BRAF down-regulated expression of endoplasmic reticulum (ER) stress proteins, and reduced unfolded protein response activation, while DN mutations increased both. Finally, a bortezomib (BTZ)/MEK inhibitor combination showed enhanced activity in vivo specifically in CA-NRAS models. Taken together, the data support the hypothesis that activating MAPK pathway mutations enhance PI resistance by increasing proteasome capacity, and provide a rationale for targeting such patients with PI/RAF or PI/MEK inhibitor combinations. Moreover, they argue these mutations promote myeloma survival by reducing cellular stress, thereby distancing plasma cells from the apoptotic threshold, potentially explaining their high frequency in myeloma.
['Shirazi|Fazal|F|', 'Jones|Richard J|RJ|', 'Singh|Ram K|RK|0000-0002-7654-1212', 'Zou|Jianxuan|J|', 'Kuiatse|Isere|I|', 'Berkova|Zuzana|Z|0000-0001-8196-6517', 'Wang|Hua|H|', 'Lee|Hans C|HC|', 'Hong|Samuel|S|', 'Dick|Larry|L|', 'Chattopadhyay|Nibedita|N|', 'Orlowski|Robert Z|RZ|']
D017209:Apoptosis D000069286:Bortezomib D059865:Endoplasmic Reticulum Stress D020558:GTP Phosphohydrolases D006801:Humans D008565:Membrane Proteins D009101:Multiple Myeloma D009154:Mutation D046988:Proteasome Endopeptidase Complex D047428:Protein Kinase Inhibitors D048493:Proto-Oncogene Proteins B-raf D016283:Proto-Oncogene Proteins p21(ras)
KRAS BRAF proteasome capacity NRAF proteasome inhibitor sensitivity