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: Suzhao Li, Gianluca Fossati, Carlo Marchetti, Daniela Modena, Pietro Pozzi, Leonid L Reznikov, Maria Luisa Moras, Tania Azam, Antonio Abbate, Paolo Mascagni, Charles A Dinarello
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 290 Issue 4 Pg. 2368-78 (Jan 23 2015) ISSN: 1083-351X [Electronic] United States
• PMID: 25451941 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Chemical References

• Cytokines
• Histone Deacetylase Inhibitors
• Interleukin-1alpha
• Interleukin-1beta
• Interleukin-6
• Lipopolysaccharides
• RNA, Messenger
• Repressor Proteins
• Tumor Necrosis Factor-alpha
• Caspase 3
• Caspase 7
• Caspase 1
• HDAC8 protein, human
• Histone Deacetylases

Topics

• Animals
• Apoptosis
• Candida (metabolism)
• Caspase 1 (metabolism)
• Caspase 3 (metabolism)
• Caspase 7 (metabolism)
• Cells, Cultured
• Cytokines (metabolism)
• Gene Expression Regulation, Enzymologic
• Histone Deacetylase Inhibitors (chemistry)
• Histone Deacetylases (metabolism)
• Humans
• Inflammation
• Inhibitory Concentration 50
• Interleukin-1alpha (metabolism)
• Interleukin-1beta (metabolism)
• Interleukin-6 (metabolism)
• Leukocytes, Mononuclear (cytology, metabolism)
• Lipopolysaccharides (chemistry)
• Mice
• Mice, Inbred C57BL
• Monocytes (cytology)
• RNA, Messenger (metabolism)
• Real-Time Polymerase Chain Reaction
• Repressor Proteins (antagonists & inhibitors, metabolism)
• Tumor Necrosis Factor-alpha (metabolism)