Inhibiting epigenetic enzymes to improve atherogenic macrophage functions

Bossche, Jan Van den; Neele, Annette E.; Hoeksema, Marten A.; Heij, Femke De; Boshuizen, Margit; Velden, Saskia van der; Boer, Vincent C.J. de; Reedquist, Kris A.; Winther, Menno P.J. de

doi:10.1016/j.bbrc.2014.11.029

Cited by 59 publications

(76 citation statements)

References 33 publications

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“…The HDAC3/6i used in these studies effectively targets both HDAC3 and HDAC6 23. HDAC3 has previously been identified as a key epigenetic modulator of inflammatory activation of murine macrophages and human PBMCs 8…”

Section: Resultsmentioning

confidence: 99%

“…25 26 Also, HDAC6 inhibition was shown to inhibit proinflammatory TNF-α and IL-6 cytokines in lipopolysaccharide (LPS)-stimulated THP-1 cells 27. To determine whether HDAC3 or HDAC6 might be responsible for the transcriptional changes observed with HDAC3/6i, we made use of an additional inhibitor specific for HDAC6 23. In initial experiments, we assessed the concentration of HDAC6i (1 µM) which induced a similar degree of tubulin acetylation as to HDAC3/6i (figure 3A) and had no effect on FLS viability (see online supplementary figure S2).…”

Section: Resultsmentioning

confidence: 99%

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Histone deacetylase 3 regulates the inflammatory gene expression programme of rheumatoid arthritis fibroblast-like synoviocytes

et al. 2016

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ObjectivesNon-selective histone deacetylase (HDAC) inhibitors (HDACi) have demonstrated anti-inflammatory properties in both in vitro and in vivo models of rheumatoid arthritis (RA). Here, we investigated the potential contribution of specific class I and class IIb HDACs to inflammatory gene expression in RA fibroblast-like synoviocytes (FLS).MethodsRA FLS were incubated with pan-HDACi (ITF2357, givinostat) or selective HDAC1/2i, HDAC3/6i, HDAC6i and HDAC8i. Alternatively, FLS were transfected with HDAC3, HDAC6 or interferon (IFN)-α/β receptor alpha chain (IFNAR1) siRNA. mRNA expression of interleukin (IL)-1β-inducible genes was measured by quantitative PCR (qPCR) array and signalling pathway activation by immunoblotting and DNA-binding assays.ResultsHDAC3/6i, but not HDAC1/2i and HDAC8i, significantly suppressed the majority of IL-1β-inducible genes targeted by pan-HDACi in RA FLS. Silencing of HDAC3 expression reproduced the effects of HDAC3/6i on gene regulation, contrary to HDAC6-specific inhibition and HDAC6 silencing. Screening of the candidate signal transducers and activators of transcription (STAT)1 transcription factor revealed that HDAC3/6i abrogated STAT1 Tyr701 phosphorylation and DNA binding, but did not affect STAT1 acetylation. HDAC3 activity was required for type I IFN production and subsequent STAT1 activation in FLS. Suppression of type I IFN release by HDAC3/6i resulted in reduced expression of a subset of IFN-dependent genes, including the chemokines CXCL9 and CXCL11.ConclusionsInhibition of HDAC3 in RA FLS largely recapitulates the effects of pan-HDACi in suppressing inflammatory gene expression, including type I IFN production in RA FLS. Our results identify HDAC3 as a potential therapeutic target in the treatment of RA and type I IFN-driven autoimmune diseases.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Histone deacetylase 3 regulates the inflammatory gene expression programme of rheumatoid arthritis fibroblast-like synoviocytes

et al. 2016

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“…Additionally, apoptosis assays (by Annexin-V/PI staining), phagocytosis assays (with fluorescent latex beads and/or pHrodo E. coli) and foam cell formation assays (by DiI-oxLDL uptake, LipidTOX neutral lipid staining) can be performed to evaluate macrophage functionality 14 . Additionally, extracellular flux analysis can be performed to measure bioenergetics profiles of polarized macrophages as a new and alternative functional readout.…”

Section: Discussionmentioning

confidence: 99%

Metabolic Characterization of Polarized M1 and M2 Bone Marrow-derived Macrophages Using Real-time Extracellular Flux Analysis

Bossche

Baardman

Winther

2015

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Citation: Van den Bossche, J., Baardman, J., de Winther, M.P. Metabolic Characterization of Polarized M1 and M2 Bone Marrow-derived Macrophages Using Real-time Extracellular Flux Analysis. J. Vis. Exp. (105), e53424, doi:10.3791/53424 (2015). AbstractSpecific metabolic pathways are increasingly being recognized as critical hallmarks of macrophage subsets. While LPS-induced classically activated M1 or M (LPS) macrophages are pro-inflammatory, IL-4 induces alternative macrophage activation and these so-called M2 or M support resolution of inflammation and wound healing. Recent evidence shows the crucial role of metabolic reprogramming in the regulation of M1 and M2 macrophage polarization.In this manuscript, an extracellular flux analyzer is applied to assess the metabolic characteristics of naive, M1 and M2 polarized mouse bone marrow-derived macrophages. This instrument uses pH and oxygen sensors to measure the extracellular acidification rate (ECAR) and oxygen consumption rate (OCR), which can be related to glycolytic and mitochondrial oxidative metabolism. As such, both glycolysis and mitochondrial oxidative metabolism can be measured in real-time in one single assay.Using this technique, we demonstrate here that inflammatory M1 macrophages display enhanced glycolytic metabolism and reduced mitochondrial activity. Conversely, anti-inflammatory M2 macrophages show high mitochondrial oxidative phosphorylation (OXPHOS) and are characterized by an enhanced spare respiratory capacity (SRC).The presented functional assay serves as a framework to investigate how particular cytokines, pharmacological compounds, gene knock outs or other interventions affect the macrophage's metabolic phenotype and inflammatory status.

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“…In primary macrophages however, the expression of the oxLDL-receptors CD36 and Msr1 is not increased upon Hdac inhibition (Van den Bossche et al, 2014a). Conversely, we and others observed increased histone acetylation and gene expression of the efflux mediators ATPbinding cassette transporter (Abca1) and ATP-binding cassette sub-family G member 1 (Abcg11) in macrophages (Ku et al, 2012;Van den Bossche et al, 2014a;Xu et al, 2011) So in contrast to what happens in macrophage cell lines, Hdac inhibition appears to have no detrimental effects on foam cell formation in primary macrophages and could even be beneficial. Hence, the observed increased atherosclerotic lesion formation upon whole-body Hdac inhibition is probably not due to increased lipid loading by macrophages, but might be explained by effects of TSA on other (non-immune) cells.…”

Section: Hdac Inhibition As Atherosclerosis Therapy?mentioning

confidence: 62%

“…We recently demonstrated that broad-spectrum Hdac inhibition has beneficial anti atherogenic effects as it partly inhibits inflammatory macrophage activation and blunts apoptosis, without augmenting foam cell formation in primary macrophages (Van den Bossche et al, 2014a). Yet, broad-spectrum Hdac inhibitors have contra-indications that prevent their direct use in atherosclerosis therapy.…”

Section: Hdac Inhibition As Atherosclerosis Therapy?mentioning

confidence: 97%