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A novel histone deacetylase inhibitor prevents IL-1beta induced metabolic dysfunction in pancreatic beta-cells.

Susick L, Senanayake T, Veluthakal R, Woster PM, Kowluru A - J. Cell. Mol. Med. (2009)

Bottom Line: Further, these compounds exerted no toxic effects on metabolic cell viability in these cells.Together, we provide evidence for a novel HDAC inhibitor with a significant potential to prevent IL-1beta-mediated effects on isolated beta-cells.Potential implications of these findings in the development of novel therapeutics to prevent deleterious effects of cytokines and the onset of autoimmune diabetes are discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, Eugene Applebaum College of Pharmacy and Health Sciences, Wayne State University, Detroit, MI 48201, USA.

ABSTRACT
The histone deacetylase (HDAC) inhibitor trichostatin A (TSA) has recently been shown to inhibit deleterious effects of cytokines on beta-cells, but it is unable to protect beta-cells from death due to its own cytotoxicity. Herein, we investigated novel HDAC inhibitors for their cytoprotective effects against IL-1beta-induced damage to isolated beta-cells. We report that three novel compounds (THS-73-44, THS-72-5 and THS-78-5) significantly inhibited HDAC activity and increased the acetylation of histone H4 in isolated beta-cells. Further, these compounds exerted no toxic effects on metabolic cell viability in these cells. However, among the three compounds tested, only THS-78-5 protected against IL-1beta-mediated loss in beta-cell viability. THS-78-5 was also able to attenuate IL-1beta-induced inducible nitric oxide synthase expression and subsequent NO release. Our data also indicate that the cytoprotective properties of THS-78-5 against IL-1beta-mediated effects may, in part, be due to inhibition of IL-1beta-induced transactivation of nuclear factor kappaB (NF-kappaB) in these cells. Together, we provide evidence for a novel HDAC inhibitor with a significant potential to prevent IL-1beta-mediated effects on isolated beta-cells. Potential implications of these findings in the development of novel therapeutics to prevent deleterious effects of cytokines and the onset of autoimmune diabetes are discussed.

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Novel HDAC inhibitors increase the acetylation of histone H4 in INS cells: (A) INS cells were incubated in the presence or absence of THS‐73–44 (10 μM), THS‐72–5 (10 μM) or THS‐78–5 (10 μM) for 24 hrs. Lysates were separated by SDS‐PAGE, transferred onto nitrocellulose membranes and subjected to immunoblot analysis with an antibody directed against acetylated H4, total H4 or actin, followed by incubation with an HRP conjugated secondary antibody for 1 hr. Immune complexes were detected using an ECL kit. A representative blot from three experiments yielding similar results is shown. (B) Relative intensities of the acetylated H4 bands were quantified by densitometry and plotted as percent of acetylated H4 in the presence of diluent alone. Data are mean ± SEM from three independent experiments. *P < 0.05 versus control cells.
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f2: Novel HDAC inhibitors increase the acetylation of histone H4 in INS cells: (A) INS cells were incubated in the presence or absence of THS‐73–44 (10 μM), THS‐72–5 (10 μM) or THS‐78–5 (10 μM) for 24 hrs. Lysates were separated by SDS‐PAGE, transferred onto nitrocellulose membranes and subjected to immunoblot analysis with an antibody directed against acetylated H4, total H4 or actin, followed by incubation with an HRP conjugated secondary antibody for 1 hr. Immune complexes were detected using an ECL kit. A representative blot from three experiments yielding similar results is shown. (B) Relative intensities of the acetylated H4 bands were quantified by densitometry and plotted as percent of acetylated H4 in the presence of diluent alone. Data are mean ± SEM from three independent experiments. *P < 0.05 versus control cells.

Mentions: It is well established that chronic exposure of pancreatic β‐cells to cytokines (e.g. IL‐1β) culminates in β‐cell death by either necrosis or apoptosis [23, 24, 25]. As stated in the Introduction, recent studies have clearly demonstrated that hypoacetylation of specific histones (e.g. histone H4) favours the induction of iNOS and subsequent NO release in isolated β‐cells in the presence of cytokines (see introduction). In the present study, we investigated the potential beneficial effects of three novel HDAC inhibitors (Fig. 1) against IL‐1β‐mediated effects in pancreatic β‐cells. At the outset, we investigated regulatory effects of these three inhibitors (Fig. 1) on histone H4 acetylation in INS cells. Data in Figure 2A and B indicate that of the three compounds tested, THS‐78–5 exerted maximal effects on histone H4 acetylation (610% of control). THS‐73–44 and THS‐72–5 demonstrated increases in acetylation of histone H4 of 160% and 440% of control, respectively.


A novel histone deacetylase inhibitor prevents IL-1beta induced metabolic dysfunction in pancreatic beta-cells.

Susick L, Senanayake T, Veluthakal R, Woster PM, Kowluru A - J. Cell. Mol. Med. (2009)

Novel HDAC inhibitors increase the acetylation of histone H4 in INS cells: (A) INS cells were incubated in the presence or absence of THS‐73–44 (10 μM), THS‐72–5 (10 μM) or THS‐78–5 (10 μM) for 24 hrs. Lysates were separated by SDS‐PAGE, transferred onto nitrocellulose membranes and subjected to immunoblot analysis with an antibody directed against acetylated H4, total H4 or actin, followed by incubation with an HRP conjugated secondary antibody for 1 hr. Immune complexes were detected using an ECL kit. A representative blot from three experiments yielding similar results is shown. (B) Relative intensities of the acetylated H4 bands were quantified by densitometry and plotted as percent of acetylated H4 in the presence of diluent alone. Data are mean ± SEM from three independent experiments. *P < 0.05 versus control cells.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3852426&req=5

f2: Novel HDAC inhibitors increase the acetylation of histone H4 in INS cells: (A) INS cells were incubated in the presence or absence of THS‐73–44 (10 μM), THS‐72–5 (10 μM) or THS‐78–5 (10 μM) for 24 hrs. Lysates were separated by SDS‐PAGE, transferred onto nitrocellulose membranes and subjected to immunoblot analysis with an antibody directed against acetylated H4, total H4 or actin, followed by incubation with an HRP conjugated secondary antibody for 1 hr. Immune complexes were detected using an ECL kit. A representative blot from three experiments yielding similar results is shown. (B) Relative intensities of the acetylated H4 bands were quantified by densitometry and plotted as percent of acetylated H4 in the presence of diluent alone. Data are mean ± SEM from three independent experiments. *P < 0.05 versus control cells.
Mentions: It is well established that chronic exposure of pancreatic β‐cells to cytokines (e.g. IL‐1β) culminates in β‐cell death by either necrosis or apoptosis [23, 24, 25]. As stated in the Introduction, recent studies have clearly demonstrated that hypoacetylation of specific histones (e.g. histone H4) favours the induction of iNOS and subsequent NO release in isolated β‐cells in the presence of cytokines (see introduction). In the present study, we investigated the potential beneficial effects of three novel HDAC inhibitors (Fig. 1) against IL‐1β‐mediated effects in pancreatic β‐cells. At the outset, we investigated regulatory effects of these three inhibitors (Fig. 1) on histone H4 acetylation in INS cells. Data in Figure 2A and B indicate that of the three compounds tested, THS‐78–5 exerted maximal effects on histone H4 acetylation (610% of control). THS‐73–44 and THS‐72–5 demonstrated increases in acetylation of histone H4 of 160% and 440% of control, respectively.

Bottom Line: Further, these compounds exerted no toxic effects on metabolic cell viability in these cells.Together, we provide evidence for a novel HDAC inhibitor with a significant potential to prevent IL-1beta-mediated effects on isolated beta-cells.Potential implications of these findings in the development of novel therapeutics to prevent deleterious effects of cytokines and the onset of autoimmune diabetes are discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, Eugene Applebaum College of Pharmacy and Health Sciences, Wayne State University, Detroit, MI 48201, USA.

ABSTRACT
The histone deacetylase (HDAC) inhibitor trichostatin A (TSA) has recently been shown to inhibit deleterious effects of cytokines on beta-cells, but it is unable to protect beta-cells from death due to its own cytotoxicity. Herein, we investigated novel HDAC inhibitors for their cytoprotective effects against IL-1beta-induced damage to isolated beta-cells. We report that three novel compounds (THS-73-44, THS-72-5 and THS-78-5) significantly inhibited HDAC activity and increased the acetylation of histone H4 in isolated beta-cells. Further, these compounds exerted no toxic effects on metabolic cell viability in these cells. However, among the three compounds tested, only THS-78-5 protected against IL-1beta-mediated loss in beta-cell viability. THS-78-5 was also able to attenuate IL-1beta-induced inducible nitric oxide synthase expression and subsequent NO release. Our data also indicate that the cytoprotective properties of THS-78-5 against IL-1beta-mediated effects may, in part, be due to inhibition of IL-1beta-induced transactivation of nuclear factor kappaB (NF-kappaB) in these cells. Together, we provide evidence for a novel HDAC inhibitor with a significant potential to prevent IL-1beta-mediated effects on isolated beta-cells. Potential implications of these findings in the development of novel therapeutics to prevent deleterious effects of cytokines and the onset of autoimmune diabetes are discussed.

Show MeSH
Related in: MedlinePlus