UHRF1 epigenetically orchestrates smooth muscle cell plasticity in arterial disease
Leonardo Elia, … , Gianluigi Condorelli, Manuela Quintavalle
Leonardo Elia, … , Gianluigi Condorelli, Manuela Quintavalle
Published March 20, 2018
Citation Information: J Clin Invest. 2018;128(6):2473-2486. https://doi.org/10.1172/JCI96121.
View: Text | PDF
Research Article Cell biology Vascular biology

UHRF1 epigenetically orchestrates smooth muscle cell plasticity in arterial disease

Abstract

Adult vascular smooth muscle cells (VSMCs) dedifferentiate in response to extracellular cues such as vascular damage and inflammation. Dedifferentiated VSMCs are proliferative, migratory, less contractile, and can contribute to vascular repair as well as to cardiovascular pathologies such as intimal hyperplasia/restenosis in coronary artery and arterial aneurysm. We here demonstrate the role of ubiquitin-like containing PHD and RING finger domains 1 (UHRF1) as an epigenetic master regulator of VSMC plasticity. UHRF1 expression correlated with the development of vascular pathologies associated with modulation of noncoding RNAs, such as microRNAs. miR-145 — pivotal in regulating VSMC plasticity, which is reduced in vascular diseases — was found to control Uhrf1 mRNA translation. In turn, UHRF1 triggered VSMC proliferation, directly repressing promoters of cell-cycle inhibitor genes (including p21 and p27) and key prodifferentiation genes via the methylation of DNA and histones. Local vascular viral delivery of Uhrf1 shRNAs or Uhrf1 VSMC-specific deletion prevented intimal hyperplasia in mouse carotid artery and decreased vessel damage in a mouse model of aortic aneurysm. Our study demonstrates the fundamental role of Uhrf1 in regulating VSMC phenotype by promoting proliferation and dedifferentiation. UHRF1 targeting may hold therapeutic potential in vascular pathologies.

Authors

Leonardo Elia, Paolo Kunderfranco, Pierluigi Carullo, Marco Vacchiano, Floriana Maria Farina, Ignacio Fernando Hall, Stefano Mantero, Cristina Panico, Roberto Papait, Gianluigi Condorelli, Manuela Quintavalle

×

Figure 2

miR-145–mediated regulation of Uhrf1.

Options: View larger image (or click on image) Download as PowerPoint
miR-145–mediated regulation of Uhrf1. (A) Putative binding sites for miR...
(A) Putative binding sites for miR-143 and miR-145 on the 3′ UTR of Uhrf1. (B) Uhrf1 expression in aortas of miR-143– and miR-145–KO mice, measured by RT-qPCR. (C) Uhrf1 expression measured by RT-qPCR in 3 different preparations of VSMCs isolated from miR-143– and miR-145–KO mice, cultured in medium with 10% FBS. (D and E) Representative RT-qPCR RNA analysis and immunoblot of the target gene UHRF1 in miR-143– and miR-145–KO VSMCs transduced with adenovirus expressing miR-208, miR-145, or miR-143 and grown in medium with 10% FBS. (F) Luciferase reporter assay on A7r5 cells stably expressing miR-145 (Ctr, cells transduced with the empty vector) with a Renilla reporter gene linked to the WT or mutated (mt) Uhrf1 3′ UTR. (G) Uhrf1 expression in WT VSMCs transduced with a lentivirus expressing miR-145 (Ctr, cells transduced with the empty vector and cells cultured in medium with 10% FBS). (H) Uhrf1 expression in WT VSMCs transduced with lentivirus expressing sponge sequences (Decoys) targeting miR-143 or miR-145 and empty vector, as measured by RT-qPCR (cells cultured in medium with 10% FBS). (I) Uhrf1 expression in WT VSMCs transduced with a lentivirus expressing miR-145 and treated with PDGF-BB (25 ng/ml) (Ctr, cells treated with vehicle). (J) Uhrf1 expression in WT VSMCs transfected with an anti–miR-145 LNA (i145) oligo and treated with TGF-β (10 ng/ml) (SCR, cells treated with scrambled oligo). All results are the average of at least 3 independent experiments and error bars indicate SD. To compare means, an unpaired 2-tailed Student’s t test was used for B, C, D, G, and J, whereas 1-way ANOVA with Tukey’s multiple comparisons t test was used for F, H, and I. #P < 0.05. Adjusted P value is shown in F, H, and I.