Pan-Raf co-operates with PI3K-dependent signalling and critically contributes to myeloma cell survival independently of mutated RAS

Leukemia. 2017 Apr;31(4):922-933. doi: 10.1038/leu.2016.264. Epub 2016 Sep 30.

Abstract

Direct therapeutic targeting of oncogenic RAS is currently still impossible due to lack of suitable pharmacological inhibitors. Because specific blockade of druggable RAS effectors might represent an alternative treatment approach, we evaluated the role of the Raf complex for multiple myeloma (MM) pathobiology. We found frequent overexpression of the Raf isoforms (A-, B- and C-Raf) and downstream activation of MEK1,2/ERK1,2 in MM cells. Concomitant inhibition of all Raf isoforms (pan-Raf inhibition) by RNAi or pharmacological inhibitors was required to strongly induce apoptosis in human MM cell lines (HMCLs), in primary MM cells in vitro, and in a syngeneic MM mouse model in vivo. The anti-MM effect of pan-Raf inhibition did not correlate with the RAS mutation status, and functionally appeared to involve both MEK-dependent and -independent mechanisms. Furthermore, transcriptome analyses revealed that pan-Raf activity affects PI3K-dependent signalling, thus highlighting a functional link between the RAS/Raf and PI3K/mTOR/Akt pro-survival pathways. Accordingly, pharmacological inhibition of PI3K strongly enhanced the anti-MM effect of pan-Raf inhibition in MM cell lines and in primary MM cells in vitro and in vivo. Concomitant pan-Raf/PI3K inhibition was also effective in carfilzomib- and lenalidomide-resistant MM models underscoring that this attractive therapeutic anti-MM strategy is suitable for immediate clinical translation.

MeSH terms

  • Apoptosis / genetics
  • Cell Line, Tumor
  • Cell Survival / genetics
  • Drug Resistance, Neoplasm
  • Enzyme Activation
  • Gene Expression
  • Gene Knockdown Techniques
  • Humans
  • Isoenzymes
  • Lenalidomide
  • MAP Kinase Signaling System / drug effects
  • Multiple Myeloma / genetics*
  • Multiple Myeloma / metabolism*
  • Mutation*
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphorylation / drug effects
  • Protein Binding
  • Protein Kinase Inhibitors / pharmacology
  • Proto-Oncogene Proteins B-raf / antagonists & inhibitors
  • Proto-Oncogene Proteins B-raf / genetics
  • Proto-Oncogene Proteins B-raf / metabolism*
  • RNA Interference
  • RNA, Small Interfering / genetics
  • Signal Transduction*
  • TOR Serine-Threonine Kinases / metabolism
  • Thalidomide / analogs & derivatives
  • Thalidomide / pharmacology
  • ras Proteins / genetics*

Substances

  • Isoenzymes
  • Protein Kinase Inhibitors
  • RNA, Small Interfering
  • Thalidomide
  • Proto-Oncogene Proteins B-raf
  • TOR Serine-Threonine Kinases
  • ras Proteins
  • Lenalidomide