BH3 mimetics potentiate pro-apoptotic activity of encorafenib in BRAFV600E melanoma cells

Cancer Lett. 2021 Feb 28:499:122-136. doi: 10.1016/j.canlet.2020.11.036. Epub 2020 Nov 28.

Abstract

BRAFV600- and MEK1/2-targeting therapies rarely produce durable response in melanoma patients. We investigated five BRAFV600E melanoma cell lines derived from drug-naïve tumor specimens to assess cell death response to encorafenib (Braftovi), a recently FDA-approved BRAFV600 inhibitor. Drug-naïve cell lines (i) did not harbor damaging alterations in genes encoding core apoptotic machinery, but they differed in (ii) mitochondrial priming as demonstrated by whole-cell BH3 profiling, and (iii) levels of selected anti-apoptotic proteins. Encorafenib modulated the balance between apoptosis-regulating proteins as it upregulated BIM and BMF, and attenuated NOXA, but did not affect the levels of pro-survival proteins except for MCL-1 and BCL-XL in selected cell lines. Induction of apoptosis could be predicted using Dynamic BH3 profiling. The extent of apoptosis was dependent on both (i) cell-intrinsic proximity to the apoptotic threshold (initial mitochondrial priming) and (ii) the abundance of encorafenib-induced BIM (iBIM; drug-induced change in priming). While co-inhibition of MCL-1 and BCL-XL/BCL-2 was indispensable for apoptosis in drug-naïve cells, encorafenib altered cell dependence to MCL-1, and reliance on BCL-XL/BCL-2 was additionally found in cell lines that were highly primed to apoptosis by encorafenib. This translated into robust apoptosis when encorafenib was combined with selective BH3 mimetics. Our study provides a mechanistic insight into the role of proteins from the BCL-2 family in melanoma cell response to targeted therapy, and presents preclinical evidence that (i) MCL-1 is a druggable target to potentiate encorafenib activity, whereas (ii) pharmacological inhibition of BCL-XL/BCL-2 might be relevant but only for a narrow group of encorafenib-treated patients.

Keywords: BH3 profiling; BRAF inhibitor; MCL-1 inhibitor; Melanoma; Targeted therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Combined Chemotherapy Protocols / pharmacology*
  • Antineoplastic Combined Chemotherapy Protocols / therapeutic use
  • Apoptosis / drug effects
  • Carbamates / pharmacology*
  • Carbamates / therapeutic use
  • Cell Line, Tumor
  • Drug Screening Assays, Antitumor
  • Drug Synergism
  • Humans
  • Melanoma / drug therapy*
  • Melanoma / genetics
  • Melanoma / pathology
  • Mutation
  • Myeloid Cell Leukemia Sequence 1 Protein / antagonists & inhibitors
  • Myeloid Cell Leukemia Sequence 1 Protein / metabolism
  • Peptide Fragments / genetics
  • Peptide Fragments / pharmacology*
  • Peptide Fragments / therapeutic use
  • Protein Interaction Domains and Motifs / genetics
  • Proto-Oncogene Proteins B-raf / antagonists & inhibitors
  • Proto-Oncogene Proteins B-raf / genetics
  • Proto-Oncogene Proteins c-bcl-2 / antagonists & inhibitors
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Skin Neoplasms / drug therapy*
  • Skin Neoplasms / genetics
  • Skin Neoplasms / pathology
  • Sulfonamides / pharmacology*
  • Sulfonamides / therapeutic use
  • bcl-2-Associated X Protein / genetics
  • bcl-X Protein / antagonists & inhibitors
  • bcl-X Protein / metabolism

Substances

  • BAX protein, human
  • BCL2 protein, human
  • BCL2L1 protein, human
  • Carbamates
  • MCL1 protein, human
  • Myeloid Cell Leukemia Sequence 1 Protein
  • Peptide Fragments
  • Proto-Oncogene Proteins c-bcl-2
  • Sulfonamides
  • bcl-2-Associated X Protein
  • bcl-X Protein
  • encorafenib
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf