Trichostatin A, a histone deacetylase inhibitor suppresses NADPH Oxidase 4-Derived Redox Signalling and Angiogenesis.
Histone deacetylase (HDAC) inhibitors are known to suppress abnormal development of blood vessels. Angiogenic activity in endothelial cells depends upon NADPH oxidase 4 (Nox4)-dependent redox signalling. We set out to study whether the HDAC inhibitor trichostatin A (TSA) affects Nox4 expression and angiogenesis. Nox4 expression was measured by real time PCR and Western blot analysis in endothelial cells. Hydrogen peroxide (H2 O2 ) was measured by amplex(®) red assay in endothelial cells. Nox4 was knocked down by Nox4 shRNA. In vitro angiogenic activities such migration and tubulogenesis were assessed using wound healing and Matrigel assays, respectively. In vivo angiogenic activity was assessed using subcutaneous sponge assay in C57Bl/6 and Nox4-deficient mice. Trichostatin A reduced Nox4 expression in a time- and concentration-dependent manner. Both TSA and Nox4 silencing decreased Nox4 protein and H2 O2 . Mechanistically, TSA reduced expression of Nox4 via ubiquitination of p300- histone acetyltransferase (p300-HAT). Thus, blocking of the ubiquitination pathway using an inhibitor of ubiquitin-activating enzyme E1 (PYR-41) prevented TSA inhibition of Nox4 expression. Trichostatin A also reduced migration and tube formation, and these effects were not observed in Nox4-deficient endothelial cells. Finally, transforming growth factor beta1 (TGFβ1) enhanced angiogenesis in sponge model in C57BL/6 mice. This response to TGFβ1 was substantially reduced in Nox4-deficient mice. Similarly intraperitoneal infusion of TSA (1 mg/kg) also suppressed TGFβ1-induced angiogenesis in C57BL/6 mice. Trichostatin A reduces Nox4 expression and angiogenesis via inhibition of the p300-HAT-dependent pathway. This mechanism might be exploited to prevent aberrant angiogenesis in diabetic retinopathy, complicated vascular tumours and malformations.
Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital, East Melbourne, VIC, Australia.|Ophthalmology, University of Melbourne, Department of Surgery, East Melbourne, VIC, Australia.|Department of Pharmacology and Therapeutics, University of Melbourne, Melbourne, VIC, Australia.|Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.
Nora Y
NY
Hakami
Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital, East Melbourne, VIC, Australia.|Ophthalmology, University of Melbourne, Department of Surgery, East Melbourne, VIC, Australia.
Gregory J
GJ
Dusting
Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital, East Melbourne, VIC, Australia. hitesh.peshavariya@unimelb.edu.au.|Ophthalmology, University of Melbourne, Department of Surgery, East Melbourne, VIC, Australia. hitesh.peshavariya@unimelb.edu.au.
Hitesh M
HM
Peshavariya
D000818:Animals; D002196:Capillaries / Q000187:drug effects / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D006877:Hydroxamic Acids / Q000494:pharmacology*; D051379:Mice; D008954:Models, Biological; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010084:Oxidation-Reduction / Q000187:drug effects; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D059748:Proteolysis / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects*; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D054875:Ubiquitination / Q000187:drug effects; D050880:p300-CBP Transcription Factors / Q000378:metabolism; D000099224:p300-CBP-Associated Factor
D056572:Histone Deacetylase Inhibitors; D006861:Hydrogen Peroxide; D006877:Hydroxamic Acids; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; D046988:Proteasome Endopeptidase Complex; D053773:Transforming Growth Factor beta1; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor; C012589:trichostatin A; C498910:NOX4 protein, human
Nox4; TGFβ1; histone acetyltransferases; histone deacetylase inhibitor; trichostatin A