Overlapping and Divergent Actions of Structurally Distinct Histone Deacetylase Inhibitors in Cardiac Fibroblasts.
Inhibitors of zinc-dependent histone deacetylases (HDACs) profoundly affect cellular function by altering gene expression via changes in nucleosomal histone tail acetylation. Historically, investigators have employed pan-HDAC inhibitors, such as the hydroxamate trichostatin A (TSA), which simultaneously targets members of each of the three zinc-dependent HDAC classes (classes I, II, and IV). More recently, class- and isoform-selective HDAC inhibitors have been developed, providing invaluable chemical biology probes for dissecting the roles of distinct HDACs in the control of various physiologic and pathophysiological processes. For example, the benzamide class I HDAC-selective inhibitor, MGCD0103 [N-(2-aminophenyl)-4-[[(4-pyridin-3-ylpyrimidin-2-yl)amino]methyl] benzamide], was shown to block cardiac fibrosis, a process involving excess extracellular matrix deposition, which often results in heart dysfunction. Here, we compare the mechanisms of action of structurally distinct HDAC inhibitors in isolated primary cardiac fibroblasts, which are the major extracellular matrix-producing cells of the heart. TSA, MGCD0103, and the cyclic peptide class I HDAC inhibitor, apicidin, exhibited a common ability to enhance histone acetylation, and all potently blocked cardiac fibroblast cell cycle progression. In contrast, MGCD0103, but not TSA or apicidin, paradoxically increased expression of a subset of fibrosis-associated genes. Using the cellular thermal shift assay, we provide evidence that the divergent effects of HDAC inhibitors on cardiac fibroblast gene expression relate to differential engagement of HDAC1- and HDAC2-containing complexes. These findings illustrate the importance of employing multiple compounds when pharmacologically assessing HDAC function in a cellular context and during HDAC inhibitor drug development.
Division of Cardiology and Consortium for Fibrosis Research and Translation, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado (K.B.S., M.S.S., W.W.B., T.A.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical, Sciences, University of Colorado Denver, Aurora, Colorado (M.F.W.); Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts (F.F.W., E.B.H.); Department of Cancer Biology, Fox Chase Cancer Center, Philadelphia, Pennsylvania (Y.-M.K., A.J.A.); and Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts (T.M.G., J.M.H.).
Katherine B
KB
Schuetze
Division of Cardiology and Consortium for Fibrosis Research and Translation, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado (K.B.S., M.S.S., W.W.B., T.A.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical, Sciences, University of Colorado Denver, Aurora, Colorado (M.F.W.); Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts (F.F.W., E.B.H.); Department of Cancer Biology, Fox Chase Cancer Center, Philadelphia, Pennsylvania (Y.-M.K., A.J.A.); and Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts (T.M.G., J.M.H.).
Matthew S
MS
Stratton
Division of Cardiology and Consortium for Fibrosis Research and Translation, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado (K.B.S., M.S.S., W.W.B., T.A.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical, Sciences, University of Colorado Denver, Aurora, Colorado (M.F.W.); Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts (F.F.W., E.B.H.); Department of Cancer Biology, Fox Chase Cancer Center, Philadelphia, Pennsylvania (Y.-M.K., A.J.A.); and Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts (T.M.G., J.M.H.).
Weston W
WW
Blakeslee
Division of Cardiology and Consortium for Fibrosis Research and Translation, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado (K.B.S., M.S.S., W.W.B., T.A.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical, Sciences, University of Colorado Denver, Aurora, Colorado (M.F.W.); Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts (F.F.W., E.B.H.); Department of Cancer Biology, Fox Chase Cancer Center, Philadelphia, Pennsylvania (Y.-M.K., A.J.A.); and Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts (T.M.G., J.M.H.).
Michael F
MF
Wempe
Division of Cardiology and Consortium for Fibrosis Research and Translation, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado (K.B.S., M.S.S., W.W.B., T.A.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical, Sciences, University of Colorado Denver, Aurora, Colorado (M.F.W.); Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts (F.F.W., E.B.H.); Department of Cancer Biology, Fox Chase Cancer Center, Philadelphia, Pennsylvania (Y.-M.K., A.J.A.); and Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts (T.M.G., J.M.H.).
Florence F
FF
Wagner
Division of Cardiology and Consortium for Fibrosis Research and Translation, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado (K.B.S., M.S.S., W.W.B., T.A.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical, Sciences, University of Colorado Denver, Aurora, Colorado (M.F.W.); Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts (F.F.W., E.B.H.); Department of Cancer Biology, Fox Chase Cancer Center, Philadelphia, Pennsylvania (Y.-M.K., A.J.A.); and Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts (T.M.G., J.M.H.).
Edward B
EB
Holson
Division of Cardiology and Consortium for Fibrosis Research and Translation, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado (K.B.S., M.S.S., W.W.B., T.A.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical, Sciences, University of Colorado Denver, Aurora, Colorado (M.F.W.); Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts (F.F.W., E.B.H.); Department of Cancer Biology, Fox Chase Cancer Center, Philadelphia, Pennsylvania (Y.-M.K., A.J.A.); and Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts (T.M.G., J.M.H.).
Yin-Ming
YM
Kuo
Division of Cardiology and Consortium for Fibrosis Research and Translation, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado (K.B.S., M.S.S., W.W.B., T.A.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical, Sciences, University of Colorado Denver, Aurora, Colorado (M.F.W.); Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts (F.F.W., E.B.H.); Department of Cancer Biology, Fox Chase Cancer Center, Philadelphia, Pennsylvania (Y.-M.K., A.J.A.); and Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts (T.M.G., J.M.H.).
Andrew J
AJ
Andrews
Division of Cardiology and Consortium for Fibrosis Research and Translation, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado (K.B.S., M.S.S., W.W.B., T.A.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical, Sciences, University of Colorado Denver, Aurora, Colorado (M.F.W.); Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts (F.F.W., E.B.H.); Department of Cancer Biology, Fox Chase Cancer Center, Philadelphia, Pennsylvania (Y.-M.K., A.J.A.); and Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts (T.M.G., J.M.H.).
Tonya M
TM
Gilbert
Division of Cardiology and Consortium for Fibrosis Research and Translation, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado (K.B.S., M.S.S., W.W.B., T.A.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical, Sciences, University of Colorado Denver, Aurora, Colorado (M.F.W.); Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts (F.F.W., E.B.H.); Department of Cancer Biology, Fox Chase Cancer Center, Philadelphia, Pennsylvania (Y.-M.K., A.J.A.); and Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts (T.M.G., J.M.H.).
Jacob M
JM
Hooker
Division of Cardiology and Consortium for Fibrosis Research and Translation, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado (K.B.S., M.S.S., W.W.B., T.A.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical, Sciences, University of Colorado Denver, Aurora, Colorado (M.F.W.); Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts (F.F.W., E.B.H.); Department of Cancer Biology, Fox Chase Cancer Center, Philadelphia, Pennsylvania (Y.-M.K., A.J.A.); and Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts (T.M.G., J.M.H.) timothy.mckinsey@ucdenver.edu.
Timothy A
TA
McKinsey
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D005347:Fibroblasts / Q000187:drug effects* / Q000201:enzymology*; D056284:Histone Deacetylase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000201:enzymology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D056572:Histone Deacetylase Inhibitors; D056284:Histone Deacetylase 1