Reversible dual inhibitor against G9a and DNMT1 improves human iPSC derivation enhancing MET and facilitating transcription factor engagement to the genome

PLoS One. 2017 Dec 27;12(12):e0190275. doi: 10.1371/journal.pone.0190275. eCollection 2017.

Abstract

The combination of defined factors with small molecules targeting epigenetic factors is a strategy that has been shown to enhance optimal derivation of iPSCs and could be used for disease modelling, high throughput screenings and/or regenerative medicine applications. In this study, we showed that a new first-in-class reversible dual G9a/DNMT1 inhibitor compound (CM272) improves the efficiency of human cell reprogramming and iPSC generation from primary cells of healthy donors and patient samples, using both integrative and non-integrative methods. Moreover, CM272 facilitates the generation of human iPSC with only two factors allowing the removal of the most potent oncogenic factor cMYC. Furthermore, we demonstrated that mechanistically, treatment with CM272 induces heterochromatin relaxation, facilitates the engagement of OCT4 and SOX2 transcription factors to OSKM refractory binding regions that are required for iPSC establishment, and enhances mesenchymal to epithelial transition during the early phase of cell reprogramming. Thus, the use of this new G9a/DNMT reversible dual inhibitor compound may represent an interesting alternative for improving cell reprogramming and human iPSC derivation for many different applications while providing interesting insights into reprogramming mechanisms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Cellular Reprogramming*
  • Genome, Human*
  • Histocompatibility Antigens
  • Histone-Lysine N-Methyltransferase / antagonists & inhibitors*
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Real-Time Polymerase Chain Reaction
  • Repressor Proteins / antagonists & inhibitors*
  • Transcription Factors / metabolism*

Substances

  • DMAP1 protein, human
  • Histocompatibility Antigens
  • Repressor Proteins
  • Transcription Factors
  • EHMT2 protein, human
  • Histone-Lysine N-Methyltransferase

Grants and funding

This work was funded by grants from Instituto de Salud Carlos III (ISCIII) PI13/00862 to JRRM, PI13/01469 to XA, PI14/01867 to FP, CIBERONC (Co-finance with FEDER funds) CB16/12/00489 to FP, TERCEL (RD12/0019/0031) to FP, ERA-NET programs TRANSCAN-2 JTC EPICA to FP and EuroNanoMed JTC NanoGene to FP, Ministry of Economy and Competitiveness “Torres Quevedo” Subprogram (PTQ-11-04777) to JRRM and by the Fundación Bancaria Caja Navarra (70270) to JRRM. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.