CYLD mediates human pulmonary artery smooth muscle cell dysfunction in congenital heart disease‐associated pulmonary arterial hypertension

Abstract

Pulmonary arterial hypertension (PAH) is a common complication of congenital heart disease (CHD). Deubiquitinase cylindromatosis (CYLD) has been reported to significantly aggravate vascular smooth muscle cell (VSMC) phenotypic transformation, proliferation, and migration. Here, we aimed to further investigate its roles and underlying mechanisms in the CHD-PAH development. The expression of CYLD in the lung tissues from CHD-PAH patients and monocrotaline (MCT) plus aortocaval (AV)-induced PAH rats, pulmonary artery smooth muscle cells (PASMCs) from MCT-AV-induced PAH rats, and human PASMCs (HPASMCs) was evaluated. After infection with CYLD siRNA or pcNDA3.1-CYLD, the proliferation, migration, and apoptosis of HPASMCs were measured using an EdU assay, transwell and scratch wound healing assays, and flow cytometric assay, respectively. An adeno-associated virus (AAV) vector encoding CYLD was used to suppress CYLD expression by being intratracheally instilled in rats 7 days before MCT-AV treatment. The results showed that CYLD was increased in the lung tissues from CHD-PAH patients and MCT-AV-induced PAH rats, and in PASMCs from MCT-AV-induced PAH rats. The contractile-type HPASMCs expressed low levels of CYLD, while the proliferative synthetic-type HPASMCs expressed high levels of CYLD. In addition, CYLD could mediate HPASMC dysfunction, which regulated HPASMC phenotypic transformation and proliferation via the modulation of p38 and ERK activation, while CYLD regulated HPASMC migration via the modulation of p38 activation. In vivo results demonstrated that the local suppression of CYLD expression could attenuate the increased levels of PAH and its associated pulmonary vascular remodeling in MCT-AV-induced PAH rats. Collectively, these results indicated that CYLD might be a potential novel therapeutic target for the prevention of PAH and pulmonary vascular remodeling in CHD-PAH through the modulation of HPASMC dysfunction.

Protective effects of calcyclin-binding protein against pulmonary vascular remodeling in flow-associated pulmonary arterial hypertension

“…The procedures performed adhered to the tenets of the Declaration of Helsinki and were approved by the Ethics Committee of Fuwai Hospital. We enrolled 22 patients with CHD-PAH who underwent repair surgery, and classified them into reversible and irreversible groups after 1 year follow-up, as described in our previous study [17]. Lung tissues from CHD-PAH patients were collected to explore expression of CacyBP/SIP.…”

“…After a 1-week adaptation period, rats were randomly divided into three groups: (1) control (n = 6), ( 2) sham (n = 6), and (3) flow-associated PAH (n = 8). Monocrotaline (MCT, Sigma-Aldrich, USA, 60 mg/kg) injection followed by abdominal aortocaval shunt (AV) was used to generate flow-associated PAH in rats, as described previously [17,18]. Rats underwent right heart catheterization to obtain hemodynamic parameters including right ventricular systolic pressure (RVSP), pulmonary arterial systolic pressure (PASP), and mean pulmonary arterial pressure (mPAP) after 4 weeks feeding [19].…”

Mechanisms orchestrating the enzymatic activity and cellular functions of deubiquitinases

“…Deregulation of DUB function is therefore expected to profoundly perturb cellular processes. Indeed, mutation of DUB genes or dysfunction of their protein homeostasis are increasingly associated with major pathologies such as neurodegenerative diseases (USP25, UCHL1, OTUB1, and AT3) ( 176 , 177 , 178 , 179 , 180 , 181 ), cancers (BAP1, CYLD, USP46, USP28, and PSMD14) ( 32 , 182 , 183 , 184 , 185 , 186 ) and inflammation (A20, CYLD, USP7, and USP47) ( 187 , 188 , 189 , 190 , 191 ). To properly target these enzymes in the clinic, several wide-ranging questions remain to be addressed: (i) What is the full spectrum of substrates modified by each DUB?…”

Chronic Sigma 1 receptor activation alleviates right ventricular dysfunction secondary to pulmonary arterial hypertension

“…Pulmonary vascular remodeling was evaluated by determining the wall thickness percentage (WT%) = [(external diameter − internal diameter)/external diameter×100 and wall area percentage (WA%) = [(total area − luminal area)/total area] ×100 of 10 arterioles per rat. [ 33 ] All analyses were performed in a blinded manner.…”