Histone deacetylase inhibitors suppress TF-kappaB-dependent agonist-driven tissue factor expression in endothelial cells and monocytes.
Histone deacetylase inhibitors (HDACi), such as trichostatin A (TSA), can regulate gene expression by promoting acetylation of histones and transcription factors. Human tissue factor (TF) expression is partly governed by a unique, NF-kappaB-related "TF-kappaB" promoter binding site. We find that TSA and four other HDACi (apicidin, MS-275, sodium butyrate, and valproic acid) all inhibit by approximately 90% TF activity and protein level induction in human umbilical vein endothelial cells stimulated by the physiologic agonists tumor necrosis factor (TNF)-alpha, interleukin-1beta, lipopolysaccharide, and HOSCN without affecting expression of the NF-kappaB-regulated adhesion molecules ICAM-1 and E-selectin. TSA and butyrate also blunt TF induction approximately 50% in vitro in peripheral blood mononuclear cells and in vivo in thioglycolate-elicited murine peritoneal macrophages. In human umbilical vein endothelial cells, TSA attenuates by approximately 70% TNF-alpha stimulation of TF mRNA transcription without affecting that of ICAM-1. By electrophoretic mobility shift assay analyses, TNF-alpha and lipopolysaccharide induce strong p65/p50 and p65/c-Rel heterodimer binding to both NF-kappaB and TF-kappaB probes. TSA nearly abolishes TF-kappaB binding without affecting NF-kappaB binding. A chromatin immunoprecipitation assay and a promoter-luciferase reporter system confirm that TSA inhibits TF-kappaB but not NF-kappaB activation. Chromatin immunoprecipitation and small interfering RNA inhibitor studies demonstrate that HDAC3 plays a significant role in TNF-alpha-mediated TF induction. Thus, HDACi transcriptionally inhibit agonist-induced TF expression in endothelial cells and monocytes by a TF-kappaB- and HDAC3-dependent mechanism. We conclude that histone deacetylases, particularly HDAC3, play a hitherto unsuspected role in regulating TF expression and raise the possibility that HDACi might be a novel therapy for thrombotic disorders.
Sections of Hematology, Oncology, and Transplantation, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Medicine Department, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Vascular Biology Center, University of Minnesota, Minneapolis, Minnesota 55455.
Jianguo
J
Wang
Sections of Hematology, Oncology, and Transplantation, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Medicine Department, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Vascular Biology Center, University of Minnesota, Minneapolis, Minnesota 55455.
Shawn A
SA
Mahmud
Sections of Medicine Department, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Pulmonary and Critical Care, University of Minnesota, Minneapolis, Minnesota 55455.
Peter B
PB
Bitterman
Sections of Medicine Department, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Vascular Biology Center, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Cardiology, University of Minnesota, Minneapolis, Minnesota 55455.
Yuqing
Y
Huo
Sections of Hematology, Oncology, and Transplantation, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Medicine Department, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Vascular Biology Center, University of Minnesota, Minneapolis, Minnesota 55455. Electronic address: slung001@umn.edu.
Arne
A
Slungaard
D000107:Acetylation / Q000187:drug effects; D002478:Cells, Cultured; D019040:E-Selectin / Q000096:biosynthesis / Q000235:genetics; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D004790:Enzyme Induction / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis / Q000235:genetics; D053583:Interleukin-1beta / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D009000:Monocytes / Q000166:cytology / Q000378:metabolism*; D016328:NF-kappa B / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology; D020218:Response Elements / Q000502:physiology*; D013925:Thromboplastin / Q000096:biosynthesis* / Q000235:genetics; D013927:Thrombosis / Q000188:drug therapy / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D000099186:Histone Deacetylase 3
D019040:E-Selectin; D004791:Enzyme Inhibitors; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases; D006657:Histones; D018799:Intercellular Adhesion Molecule-1; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D016328:NF-kappa B; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D013925:Thromboplastin; D014409:Tumor Necrosis Factor-alpha; D000099186:Histone Deacetylase 3