Cellular plasticity is fundamental in biology and disease. Vascular smooth muscle cell (SMC) dedifferentiation (loss of contractile proteins) initiates and perpetrates vascular pathologies such as restenosis. Contractile gene expression is governed by the master transcription factor, serum response factor (SRF). Unlike other histone deacetylases, histone deacetylase 6 (HDAC6) primarily resides in the cytosol. Whether HDAC6 regulates SRF nuclear activity was previously unknown in any cell type. This study found that selective inhibition of HDAC6 with tubastatin A preserved the contractile protein (alpha-smooth muscle actin) that was otherwise diminished by platelet-derived growth factor-BB. Tubastatin A also enhanced SRF transcriptional (luciferase) activity, and this effect was confirmed by HDAC6 knockdown. Interestingly, HDAC6 inhibition increased acetylation and total protein of myocardin-related transcription factor A (MRTF-A), a transcription co-activator known to translocate from the cytosol to the nucleus, thereby activating SRF. Consistently, HDAC6 co-immunoprecipitated with MRTF-A. In vivo studies showed that tubastatin A treatment of injured rat carotid arteries mitigated neointimal lesion, which is known to be formed largely by dedifferentiated SMCs. This report is the first to show HDAC6 regulation of the MRTF-A/SRF axis and SMC plasticity, thus opening a new perspective for interventions of vascular pathologies.
Keywords: DMEM, Dulbecco’s modified Eagle’s medium; DNA, deoxyribonucleic acid; EEL, external elastic lamina; FBS, fetal bovine serum; HDAC, histone deacetylase; HDAC6; IEL, internal elastic lamina; IH, intimal hyperplasia; IgG, immunoglobulin G; MMP, matrix metalloproteinase; MRTF-A; MRTF-A, myocardin-related transcription factor A; PDGF-BB, platelet-derived growth factor-BB; SMA, smooth muscle actin; SMC, vascular smooth muscle cell; SMHC, smooth muscle myosin heavy chain; SRF; SRF, serum response factor; TNF, tumor necrosis factor; TSA, trichostatin A; dedifferentiation; siRNA, small interfering ribonucleic acid; vascular smooth muscle cell.