Essential Role of DNA Methyltransferase 1-mediated Transcription of Insulin-like Growth Factor 2 in Resistance to Histone Deacetylase Inhibitors

Clin Cancer Res. 2017 Mar 1;23(5):1299-1311. doi: 10.1158/1078-0432.CCR-16-0534. Epub 2016 Aug 31.

Abstract

Purpose: Histone deacetylase inhibitors (HDI) are promising anticancer therapies; however, drug resistance limits their efficacy. Here, we investigated the molecular mechanisms underlying HDI resistance, focusing on the mechanism of HDI-mediated induction of insulin-like growth factor 2 (IGF2) based on our previous study.Experimental Design: The methylation status of CCCTC-binding factor (CTCF)-binding sites in the IGF2/H19 imprinting control region (ICR) were determined by methylation-specific PCR and bisulfite sequencing. The effectiveness of single or combinatorial blockade of DNA methyltransferase 1 (DNMT1) and histone deacetylase (HDAC) was evaluated using cell viability assay and patient-derived tumor xenograft (PDX) model.Results: HDAC inhibition by vorinostat increased acetylated STAT3 (K685), resulting in transcriptional upregulation of DNMT1 DNMT1-mediated hypermethylation of CTCF-binding sites in the IGF2/H19 ICR decreased CTCF insulator activity, leading to a transcriptional upregulation of IGF2 and activation of the insulin-like growth factor 1 receptor (IGF-1R) pathway in cells with acquired or de novo vorinostat resistance. Strategies targeting DNMT1 diminished the IGF2 expression and potentiated vorinostat sensitivity in preclinical models of lung cancer with hypermethylation in the H19/IGF2 ICR. The degree of ICR hypermethylation correlated with vorinostat resistance in patient-derived lung tumors and in patients with hematologic malignancies.Conclusions: DNMT1-mediated transcriptional upregulation of IGF2 is a novel mechanism of resistance to HDIs, highlighting the role of epigenetic deregulation of IGF2 in HDI resistance and the potential value of the H19/IGF2 ICR hypermethylation and DNMT1 expression as predictive biomarkers in HDI-based anticancer therapies. Clin Cancer Res; 23(5); 1299-311. ©2016 AACR.

MeSH terms

  • Animals
  • CCCTC-Binding Factor / genetics
  • DNA (Cytosine-5-)-Methyltransferase 1 / genetics*
  • DNA Methylation / genetics
  • Drug Resistance, Neoplasm / genetics
  • Gene Expression Regulation, Neoplastic / drug effects
  • Hematologic Neoplasms / drug therapy*
  • Hematologic Neoplasms / genetics
  • Hematologic Neoplasms / pathology
  • Histone Deacetylase Inhibitors / adverse effects
  • Histone Deacetylases / genetics
  • Humans
  • Hydroxamic Acids / administration & dosage
  • Insulin-Like Growth Factor II / genetics*
  • Lung Neoplasms / drug therapy
  • Lung Neoplasms / genetics*
  • Lung Neoplasms / pathology
  • Lung Neoplasms / secondary
  • Mice
  • RNA, Long Noncoding / genetics*
  • STAT3 Transcription Factor / genetics
  • Vorinostat
  • Xenograft Model Antitumor Assays

Substances

  • CCCTC-Binding Factor
  • CTCF protein, human
  • H19 long non-coding RNA
  • Histone Deacetylase Inhibitors
  • Hydroxamic Acids
  • IGF2 protein, human
  • RNA, Long Noncoding
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • Vorinostat
  • Insulin-Like Growth Factor II
  • DNA (Cytosine-5-)-Methyltransferase 1
  • DNMT1 protein, human
  • Histone Deacetylases