Small molecule inhibitors and CRISPR/Cas9 mutagenesis demonstrate that SMYD2 and SMYD3 activity are dispensable for autonomous cancer cell proliferation

PLoS One. 2018 Jun 1;13(6):e0197372. doi: 10.1371/journal.pone.0197372. eCollection 2018.

Abstract

A key challenge in the development of precision medicine is defining the phenotypic consequences of pharmacological modulation of specific target macromolecules. To address this issue, a variety of genetic, molecular and chemical tools can be used. All of these approaches can produce misleading results if the specificity of the tools is not well understood and the proper controls are not performed. In this paper we illustrate these general themes by providing detailed studies of small molecule inhibitors of the enzymatic activity of two members of the SMYD branch of the protein lysine methyltransferases, SMYD2 and SMYD3. We show that tool compounds as well as CRISPR/Cas9 fail to reproduce many of the cell proliferation findings associated with SMYD2 and SMYD3 inhibition previously obtained with RNAi based approaches and with early stage chemical probes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • A549 Cells
  • Adenocarcinoma of Lung / drug therapy*
  • Adenocarcinoma of Lung / genetics
  • Adenocarcinoma of Lung / pathology
  • CRISPR-Cas Systems
  • Carcinogenesis / drug effects
  • Carcinogenesis / genetics*
  • Cell Proliferation / drug effects
  • Enzyme Inhibitors / pharmacology
  • Histone-Lysine N-Methyltransferase / antagonists & inhibitors
  • Histone-Lysine N-Methyltransferase / chemistry
  • Histone-Lysine N-Methyltransferase / genetics*
  • Humans
  • Methylation / drug effects
  • Methyltransferases / antagonists & inhibitors
  • RNA Interference
  • Small Molecule Libraries / pharmacology

Substances

  • Enzyme Inhibitors
  • Small Molecule Libraries
  • Methyltransferases
  • Histone-Lysine N-Methyltransferase
  • SMYD2 protein, human
  • SMYD3 protein, human

Grants and funding

All work described in this manuscript was funded by Epizyme Inc. (www.epizyme.com) who provided support in the form of salaries for all authors and played a role in the study design, data collection and analysis, decision to publish and preparation of the manuscript. The specific roles of these authors are articulated in the author contributions’ section.