1. Academic Validation
  2. Specific inhibition of histone deacetylase 8 reduces gene expression and production of proinflammatory cytokines in vitro and in vivo

Specific inhibition of histone deacetylase 8 reduces gene expression and production of proinflammatory cytokines in vitro and in vivo

  • J Biol Chem. 2015 Jan 23;290(4):2368-78. doi: 10.1074/jbc.M114.618454.
Suzhao Li 1 Gianluca Fossati 2 Carlo Marchetti 3 Daniela Modena 2 Pietro Pozzi 2 Leonid L Reznikov 1 Maria Luisa Moras 2 Tania Azam 1 Antonio Abbate 3 Paolo Mascagni 2 Charles A Dinarello 4
Affiliations

Affiliations

  • 1 From the Department of Medicine, University of Colorado Denver, Aurora, Colorado 80045.
  • 2 Italfarmaco, S.p.A., Cinisello Balsamo 20092, Italy.
  • 3 the Department of Medicine, Virginia Commonwealth University, Richmond, Virginia 23298, and.
  • 4 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 [email protected].
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.

Keywords

Anti-inflammatory; Cell Death; Cytokine; Histone Deacetylase Inhibitor (HDAC Inhibitor) (HDI); Human Peripheral Blood Mononuclear Cells; ITF2357; ITF3056; Inflammation; Lipopolysaccharide (LPS).

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