A novel approach to detect resistance mechanisms reveals FGR as a factor mediating HDAC inhibitor SAHA resistance in B-cell lymphoma

Mol Oncol. 2016 Oct;10(8):1232-44. doi: 10.1016/j.molonc.2016.06.001. Epub 2016 Jun 9.

Abstract

Histone deacetylase (HDAC) inhibitors such as suberoylanilide hydroxamic acid (SAHA) are not commonly used in clinical practice for treatment of B-cell lymphomas, although a subset of patients with refractory or relapsed B-cell lymphoma achieved partial or complete remissions. Therefore, the purpose of this study was to identify molecular features that predict the response of B-cell lymphomas to SAHA treatment. We designed an integrative approach combining drug efficacy testing with exome and captured target analysis (DETECT). In this study, we tested SAHA sensitivity in 26 B-cell lymphoma cell lines and determined SAHA-interacting proteins in SAHA resistant and sensitive cell lines employing a SAHA capture compound (CC) and mass spectrometry (CCMS). In addition, we performed exome mutation analysis. Candidate validation was done by expression analysis and knock-out experiments. An integrated network analysis revealed that the Src tyrosine kinase Gardner-Rasheed feline sarcoma viral (v-fgr) oncogene homolog (FGR) is associated with SAHA resistance. FGR was specifically captured by the SAHA-CC in resistant cells. In line with this observation, we found that FGR expression was significantly higher in SAHA resistant cell lines. As functional proof, CRISPR/Cas9 mediated FGR knock-out in resistant cells increased SAHA sensitivity. In silico analysis of B-cell lymphoma samples (n = 1200) showed a wide range of FGR expression indicating that FGR expression might help to stratify patients, which clinically benefit from SAHA therapy. In conclusion, our comprehensive analysis of SAHA-interacting proteins highlights FGR as a factor involved in SAHA resistance in B-cell lymphoma.

Keywords: B-cell lymphoma; Drug resistance; FGR; HDAC inhibitor; SAHA.

MeSH terms

  • Cell Line, Tumor
  • Drug Resistance, Neoplasm / drug effects*
  • Gene Knockout Techniques
  • Gene Regulatory Networks / drug effects
  • Histone Deacetylase Inhibitors / pharmacology*
  • Humans
  • Hydroxamic Acids / pharmacology*
  • Lymphoma, B-Cell / pathology*
  • Mass Spectrometry
  • Mutation / genetics
  • Proto-Oncogene Proteins / metabolism*
  • Reproducibility of Results
  • Vorinostat
  • src-Family Kinases / metabolism*

Substances

  • Histone Deacetylase Inhibitors
  • Hydroxamic Acids
  • Proto-Oncogene Proteins
  • Vorinostat
  • proto-oncogene proteins c-fgr
  • src-Family Kinases