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

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-1beta 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-1beta, IL-1alpha, TNFalpha, 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-1beta was reduced by 76%, secretion of TNFalpha was reduced by 88%, and secretion of IL-6 was reduced by 61%. The intracellular levels of IL-1alpha were 30% lower. There was no evidence of cell toxicity at ITF3056 concentrations of 100-1000 nm. Gene expression of TNFalpha was markedly reduced (80%), whereas IL-6 gene expression was 40% lower. Although anti-CD3/28 and Candida stimulation of IL-1beta and TNFalpha was modestly reduced, IFNgamma production was 75% lower. Mechanistically, ITF3056 reduced the secretion of processed IL-1beta independent of inhibition of caspase-1 activity; however, synthesis of the IL-1beta precursor was reduced by 40% without significant decrease in IL-1beta mRNA levels. In mice, ITF3056 reduced LPS-induced serum TNFalpha by 85% and reduced IL-1beta by 88%. These data suggest that specific inhibition of HDAC8 results in reduced inflammation without cell toxicity