title: Specific inhibition of histone deacetylase 8 reduces gene expression and production of proinflammatory cytokines in vitro and in vivo.
abstract: ITF2357 (generic givinostat) is an orally active, hydroxamic-containing histone deacetylase (HDAC) inhibitor with broad anti-inflammatory properties, which has been used to treat children with systemic juvenile idiopathic arthritis. ITF2357 inhibits both Class I and II HDACs and reduces caspase-1 activity in human peripheral blood mononuclear cells and the secretion of IL-1β and other cytokines at 25-100 nm; at concentrations >200 nm, ITF2357 is toxic in vitro. ITF3056, an analog of ITF2357, inhibits only HDAC8 (IC50 of 285 nm). Here we compared the production of IL-1β, IL-1α, TNFα, and IL-6 by ITF2357 with that of ITF3056 in peripheral blood mononuclear cells stimulated with lipopolysaccharide (LPS), heat-killed Candida albicans, or anti-CD3/anti-CD28 antibodies. ITF3056 reduced LPS-induced cytokines from 100 to 1000 nm; at 1000 nm, the secretion of IL-1β was reduced by 76%, secretion of TNFα was reduced by 88%, and secretion of IL-6 was reduced by 61%. The intracellular levels of IL-1α were 30% lower. There was no evidence of cell toxicity at ITF3056 concentrations of 100-1000 nm. Gene expression of TNFα was markedly reduced (80%), whereas IL-6 gene expression was 40% lower. Although anti-CD3/28 and Candida stimulation of IL-1β and TNFα was modestly reduced, IFNγ production was 75% lower. Mechanistically, ITF3056 reduced the secretion of processed IL-1β independent of inhibition of caspase-1 activity; however, synthesis of the IL-1β precursor was reduced by 40% without significant decrease in IL-1β mRNA levels. In mice, ITF3056 reduced LPS-induced serum TNFα by 85% and reduced IL-1β by 88%. These data suggest that specific inhibition of HDAC8 results in reduced inflammation without cell toxicity.
authors.affiliation: From the Department of Medicine, University of Colorado Denver, Aurora, Colorado 80045.
authors.forename: Suzhao
authors.initials: S
authors.lastname: Li
authors.affiliation: Italfarmaco, S.p.A., Cinisello Balsamo 20092, Italy.
authors.forename: Gianluca
authors.initials: G
authors.lastname: Fossati
authors.affiliation: the Department of Medicine, Virginia Commonwealth University, Richmond, Virginia 23298, and.
authors.forename: Carlo
authors.initials: C
authors.lastname: Marchetti
authors.affiliation: Italfarmaco, S.p.A., Cinisello Balsamo 20092, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Modena
authors.affiliation: Italfarmaco, S.p.A., Cinisello Balsamo 20092, Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Pozzi
authors.affiliation: From the Department of Medicine, University of Colorado Denver, Aurora, Colorado 80045.
authors.forename: Leonid L
authors.initials: LL
authors.lastname: Reznikov
authors.affiliation: Italfarmaco, S.p.A., Cinisello Balsamo 20092, Italy.
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: Moras
authors.affiliation: From the Department of Medicine, University of Colorado Denver, Aurora, Colorado 80045.
authors.forename: Tania
authors.initials: T
authors.lastname: Azam
authors.affiliation: the Department of Medicine, Virginia Commonwealth University, Richmond, Virginia 23298, and.
authors.forename: Antonio
authors.initials: A
authors.lastname: Abbate
authors.affiliation: Italfarmaco, S.p.A., Cinisello Balsamo 20092, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Mascagni
authors.affiliation: From the Department of Medicine, University of Colorado Denver, Aurora, Colorado 80045, the Department of Medicine, Radboud University Medical Centre, 6525 HP Nijmegen, The Netherlands cdinare333@aol.com.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Dinarello
mesh_terms: D000818:Animals; D017209:Apoptosis; D002175:Candida / Q000378:metabolism; D020170:Caspase 1 / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D056572:Histone Deacetylase Inhibitors / Q000737:chemistry*; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D007249:Inflammation; D020128:Inhibitory Concentration 50; D053582:Interleukin-1alpha / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism; D008070:Lipopolysaccharides / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D009000:Monocytes / Q000166:cytology; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012097:Repressor Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D016207:Cytokines; D056572:Histone Deacetylase Inhibitors; D053582:Interleukin-1alpha; D053583:Interleukin-1beta; D015850:Interleukin-6; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D012097:Repressor Proteins; D014409:Tumor Necrosis Factor-alpha; D053148:Caspase 3; D053179:Caspase 7; D020170:Caspase 1; C409470:HDAC8 protein, human; D006655:Histone Deacetylases
keywords: Anti-inflammatory; Cell Death; Cytokine; Histone Deacetylase Inhibitor (HDAC Inhibitor) (HDI); Human Peripheral Blood Mononuclear Cells; ITF2357; ITF3056; Inflammation; Lipopolysaccharide (LPS)