UHRF1 epigenetically orchestrates smooth muscle cell plasticity in arterial disease
Leonardo Elia, Paolo Kunderfranco, Pierluigi Carullo, Marco Vacchiano, Floriana Maria Farina, Ignacio Fernando Hall, Stefano Mantero, Cristina Panico, Roberto Papait, Gianluigi Condorelli, Manuela Quintavalle
Leonardo Elia, Paolo Kunderfranco, Pierluigi Carullo, Marco Vacchiano, Floriana Maria Farina, Ignacio Fernando Hall, Stefano Mantero, Cristina Panico, Roberto Papait, Gianluigi Condorelli, Manuela Quintavalle
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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

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Figure 5

Uhrf1 regulates VSMC plasticity in vitro.

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Uhrf1 regulates VSMC plasticity in vitro. (A) Proliferation curve of VS...
(A) Proliferation curve of VSMCs stably expressing shSCR or shUHRF1. To determine growth curves, 2 × 104 cells/ml were plated in 6-well plates and cultured with 10% FBS. The number of viable cells was counted for 3 days. (B) Proliferation measured by BrdU incorporation in shSCR and shUHRF1 SMCs. (C) Cell-cycle analysis of shSCR- and shUHRF1-expressing SMCs. (D and E) RNA and protein analysis of different cyclins involved in VSMC phenotypic switch. (F) ChIP analysis showing UHRF1 enrichment on the Cdkn1b promoter in proliferating cells. Data are presented as mean relative enrichment over input ± SD of 3 biological repeats. (G) Results from methyl-ChIP experiments showing reduction of methylation at the promoter in the absence of Uhrf1. Data are presented as mean relative enrichment over input ± SD of 3 biological repeats. (H) ChIP showing the enrichment of H3K27me3 in control VSMCs compared with UHRF1-silenced cells. Data are presented as mean relative enrichment over input ± SD of 3 biological repeats. If not otherwise stated, the results are the average of at least 3 independent experiments and error bars indicate SD. To compare means, 1-way ANOVA with Tukey’s multiple comparisons test was used. #Adjusted P < 0.05.