Cezanne is a critical regulator of pathological arterial remodelling by targeting β-catenin signalling

Abstract

Aim Pathological arterial remodelling including neointimal hyperplasia and atherosclerosis is the main underlying cause for occluding arterial diseases. Cezanne is a novel deubiquitinating enzyme, functioning as a NF-кB negative regulator, and plays a key role in renal inflammatory response and kidney injury induced by ischemia. Here we attempted to examine its pathological role in vascular smooth muscle cell (VSMC) pathology and arterial remodelling. Methods and Results Cezanne expression levels were consistently induced by various atherogenic stimuli in VSMCs, and in remodelled arteries upon injury. Functionally, VSMCs over-expressing wild-type Cezanne, but not the mutated catalytically-inactive Cezanne (C209S), had an increased proliferative ability and mobility, while the opposite was observed in VSMCs with Cezanne knockdown. Surprisingly, we observed no significant effects of Cezanne on VSMC apoptosis, NF-κB signalling, or inflammation. RNA-sequencing and biochemical studies showed that Cezanne drives VSMC proliferation by regulating CCN family member 1 (CCN1) by targeting β-catenin for deubiquitination. Importantly, local correction of Cezanne expression in the injured arteries greatly decreased VSMC proliferation, and prevented arterial inward remodelling. Interestingly, global Cezanne gene deletion in mice led to smaller atherosclerotic plaques, but with a lower level of plaque stability. Translating, we observed a similar role for Cezanne in human VSMCs, and higher expression levels of Cezanne in human atherosclerotic lesions. Conclusion Cezanne is a key regulator of VSMC proliferation and migration in pathological arterial remodelling. Our findings have important implications for therapeutic targeting Cezanne signalling and VSMC pathology in vascular diseases. Translational Perspective In this study, we have identified the deubiquitinating enzyme Cezanne as a novel regulator in governing VSMC phenotype, injury-induced neointimal hyperplasia, and hyperlipidaemia-induced atherosclerosis. Since accumulating evidence highlights an important role for VSMC dysfunctions in many cardiovascular pathological conditions including atherosclerosis, arterial remodelling, hypertension, and stroke, local modulation of this newly identified signal axis (Cezanne/β-catenin/CCN1) could represent as a novel therapy for post-angioplasty restenosis and aforementioned diseases.

Targeted Inhibition of Matrix Metalloproteinase-8 Prevents Aortic Dissection in a Murine Model

“…As described previously [ 32 , 33 ], cell apoptosis was detected in cultured SMCs with the indicated treatments, as shown in the respective figure legend, using a commercial Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) Assay Kit (11684795910, Sigma, Dorset, UK), according to the manufacturer’s instructions. Cells were co-stained with DAPI.…”

“…C166 cells, a widely used mouse endothelial cell line, were purchased from ATCC (CRL-2581) and maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10 % FBS and 1 % penicillin/streptomycin, as per the manufacturer’s instructions. Control scramble esiRNA or Vcam1-specific esiRNA (50 nM, final concentration) were transfected into C166 cells using TransIT-X2 Transfection Reagent (Geneflow Limited, Lichfield, UK), according to the manufacturer’s instructions, and as previously described [ 33 ]. MISSION esiRNA is a heterogeneous mixture of siRNAs that all target the same mRNA sequence, resulting in highly specific and effective gene silencing.…”

“…Primary murine aortic SMCs were isolated from eight-week-old WT or MMP8_KO mice with both sexes, and routinely maintained in DMEM supplemented with 10% FBS, as described in our previous studies [ 32 , 33 , 35 , 36 , 37 ]. Aortic SMCs between Passages 3 (P3) and 8 (P8) were used in our study, as described previously [ 38 ].…”

Ferulic Acid Alleviates Atherosclerotic Plaques by Inhibiting VSMC Proliferation Through the NO/p21 Signaling pathway

“…Therefore, by inhibiting the proliferation and migration of VSMCs, the number of foam cells generated from smooth muscle cells can be reduced, which is conducive to preventing the occurrence and development of AS atherosclerotic plaques and is beneficial for the treatment of AS [8]. However, it has also been proposed that VSMC proliferation and migration play an important role in maintaining the stability of plaques and are important compensatory mechanisms of plaque injury [9]. Therefore, the advantages and disadvantages of therapeutic inhibition of VSMC proliferation and migration in AS are still very controversial, and more experimental evidence is needed.…”

Inhibition of VRK1 suppresses proliferation and migration of vascular smooth muscle cells and intima hyperplasia after injury via mTORC1/β-catenin axis

“…β-catenin is a critical regulator of the proliferation and migration of VSMCs for vascular remodeling ( 11 ). A previous study has demonstrated that VRK1 can induce the proliferation and migration of gastric carcinoma cells by targeting β-catenin ( 12 ).…”