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

Michael J Thomenius; Jennifer Totman; Darren Harvey; Lorna H Mitchell; Thomas V Riera; Kat Cosmopoulos; Alexandra R Grassian; Christine Klaus; Megan Foley; Elizabeth A Admirand; Haris Jahic; Christina Majer; Tim Wigle; Suzanne L Jacques; Jodi Gureasko; Dorothy Brach; Trupti Lingaraj; Kip West; Sherri Smith; Nathalie Rioux; Nigel J Waters; Cuyue Tang; Alejandra Raimondi; Michael Munchhof; James E Mills; Scott Ribich; Margaret Porter Scott; Kevin W Kuntz; William P Janzen; Mikel Moyer; Jesse J Smith; Richard Chesworth; Robert A Copeland; P Ann Boriack-Sjodin

doi:10.1371/journal.pone.0197372

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.

Authors

Michael J Thomenius¹, Jennifer Totman¹, Darren Harvey¹, Lorna H Mitchell¹, Thomas V Riera¹, Kat Cosmopoulos¹, Alexandra R Grassian¹, Christine Klaus¹, Megan Foley¹, Elizabeth A Admirand¹, Haris Jahic¹, Christina Majer¹, Tim Wigle¹, Suzanne L Jacques¹, Jodi Gureasko¹, Dorothy Brach¹, Trupti Lingaraj¹, Kip West¹, Sherri Smith¹, Nathalie Rioux¹, Nigel J Waters¹, Cuyue Tang¹, Alejandra Raimondi¹, Michael Munchhof¹, James E Mills¹, Scott Ribich¹, Margaret Porter Scott¹, Kevin W Kuntz¹, William P Janzen¹, Mikel Moyer¹, Jesse J Smith¹, Richard Chesworth¹, Robert A Copeland¹, P Ann Boriack-Sjodin¹

Affiliation

¹ Epizyme, Inc., Cambridge, Massachusetts, United States of America.

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.