ADAMTS8 Promotes the Development of Pulmonary Arterial Hypertension and Right Ventricular Failure

Abstract

Rationale: Pulmonary arterial hypertension (PAH) is characterized by pulmonary vascular remodeling with aberrant pulmonary artery smooth muscle cells (PASMCs) proliferation, endothelial dysfunction, and extracellular matrix remodeling. Objective: Right ventricular (RV) failure is an important prognostic factor in PAH. Thus, we need to elucidate a novel therapeutic target in both PAH and RV failure. Methods and Results: We performed microarray analysis in PASMCs from patients with PAH (PAH-PASMCs) and controls. We found a ADAMTS8 (disintegrin and metalloproteinase with thrombospondin motifs 8), a secreted protein specifically expressed in the lung and the heart, was upregulated in PAH-PASMCs and the lung in hypoxia-induced pulmonary hypertension (PH) in mice. To elucidate the role of ADAMTS8 in PH, we used vascular smooth muscle cell-specific ADAMTS8-knockout mice (ADAMTS ΔSM22 ). Hypoxia-induced PH was attenuated in ADAMTS ΔSM22 mice compared with controls. ADAMTS8 overexpression increased PASMC proliferation with downregulation of AMPK (AMP-activated protein kinase). In contrast, deletion of ADAMTS8 reduced PASMC proliferation with AMPK upregulation. Moreover, deletion of ADAMTS8 reduced mitochondrial fragmentation under hypoxia in vivo and in vitro. Indeed, PASMCs harvested from ADAMTS ΔSM22 mice demonstrated that phosphorylated DRP-1 (dynamin-related protein 1) at Ser637 was significantly upregulated with higher expression of profusion genes (Mfn1 and Mfn2) and improved mitochondrial function. Moreover, recombinant ADAMTS8 induced endothelial dysfunction and matrix metalloproteinase activation in an autocrine/paracrine manner. Next, to elucidate the role of ADAMTS8 in RV function, we developed a cardiomyocyte-specific ADAMTS8 knockout mice (ADAMTS8 ΔαMHC ). ADAMTS8 ΔαMHC mice showed ameliorated RV failure in response to chronic hypoxia. In addition, ADAMTS8 ΔαMHC mice showed enhanced angiogenesis and reduced RV ischemia and fibrosis. Finally, high-throughput screening revealed that mebendazole, which is used for treatment of parasite infections, reduced ADAMTS8 expression and cell proliferation in PAH-PASMCs and ameliorated PH and RV failure in PH rodent models. Conclusions: These results indicate that ADAMTS8 is a novel therapeutic target in PAH.

Implication of EZH2 in the Pro-Proliferative and Apoptosis-Resistant Phenotype of Pulmonary Artery Smooth Muscle Cells in PAH: A Transcriptomic and Proteomic Approach

“…In this regard, Survivin and E2F1, found to be significantly downregulated by RNA-Seq, were significantly decreased by Western blot but not by LC-MS/MS proteomics. Importantly, we found that numerous transcripts of genes already known to be implicated in the pro-proliferative and apoptosisresistant phenotype of PAH-PASMCs, such as Survivin [21], ADAMTS8 [35], PBK [20], TK1 [36], and E2F1 [37], were significantly repressed under EZH2 inhibition, which is consistent with the hypothesis that EZH2 plays a critical role in the abnormal phenotype of disease cells. Furthermore, several factors implicated in the protection of DNA damage or in DNA repair itself were diminished.…”

Identification of candidate biomarkers and pathways associated with type 1 diabetes mellitus using bioinformatics analysis

“…A previous study reported that FAP (fibroblast activation protein alpha) [205], EYA4 [206], BCL9 [207], IRF2BP2 [208], EGR3 [209], GADD45B [210], DMD (dystrophin) [211], LSR (lipolysis stimulated lipoprotein receptor) [212], DLL4 [213], SUN2 [214], SOS1 [215], PIK3CA [216], GAMT (guanidinoacetate N-methyltransferase) [217], RBM47 [218], HSP90AA1 [219], GAB1 [220], S1PR1 [221], EDNRB (endothelin receptor type B) [222], NFKBIA (NFKB inhibitor alpha) [223], GJA1 [224], GADD45G [225], PHLDA1 [226], CMPK2 [227], FIGN (fidgetin, microtubule severing factor) [228], KCNJ2 [229], ABCC9 [230], DIRAS3 [231], EPHX1 [232], RAB4A [233], UBIAD1 [234], CASQ2 [235], TTN (titin) [236], KCNH1 [237], JPH2 [238], OXGR1 [239], UCHL1 [240], SERPINA3 [241], MMP28 [242], ADAMTS2 [243], P2RY1 [244], CSF2RA [245], MYO1F [246], SELPLG (selectin P ligand) [247] and SAMHD1 [248] are expressed in cardiovascular disease, but these genes might be novel target for T1DM. MAOB (monoamine oxidase B) [249], VEGFC (vascular endothelial growth factor C) [250], DBP (D-box binding PAR bZIP transcription factor) [251], MYADM (myeloid associated differentiation marker) [252], NES (nestin) [253], SMURF1 [254], EDNRB (endothelin receptor type B) [255], MUC6 [256], TOR2A [257], TNKS (tankyrase) [258], NEDD9 [259], ASIC1 [260], ADAMTS8 [261], DYSF (dysferlin) [262], SLC26A9 [263], SLC45A3 [264] and KCNQ2 [265] contributes to the progression of hypertension, but these genes might be novel target for T1DM. Yang et al [266], Zhang et al [267] and Wang et al [268] demonstrated that SYVN1, BTG1 and CFB (complement factor B) were associated with diabetic retinopathy, but these genes mi...…”

ADAMTS proteases in cardiovascular physiology and disease

“…ADAMTS-8 expression was increased in the lungs of patients with pulmonary arterial hypertension (PAH) and in mouse/rat models of PAH [119]. In a model of hypoxia-induced PAH, mice bearing a targeted deletion of Adamts8 in pulmonary arterial smooth muscle cells (Adamts8 ΔSM22α ) showed decreased right ventricular systolic pressure and right ventricular hypertrophy compared with wild-type mice, suggesting a crucial role for ADAMTS-8 in the development of PAH [119]. Addition of recombinant ADAMTS-8 to pulmonary artery ECs seemed to exert a pro-inflammatory and pro-apoptotic role [119], similar ADAMTS-19 --Adamts19 −/− aortic regurgitation, aortic stenosis, BAV [122] royalsocietypublishing.org/journal/rsob Open Biol.…”

“…ADAMTS8 has been identified as a tumour suppressor gene in several types of cancers [164] and single nucleotide polymorphisms (SNPs) in the ADAMTS8 locus have been associated with hypertension in a genome-wide association study (GWAS) [165]. At the protein level, ADAMTS-8 is highly expressed in the lung and the heart [119,166] and, together with ADAMTS-1, -4 and -5, has been detected within human carotid lesions and advanced coronary atherosclerotic plaques [106,107]. ADAMTS-8 expression was increased in the lungs of patients with pulmonary arterial hypertension (PAH) and in mouse/rat models of PAH [119].…”

“…At the protein level, ADAMTS-8 is highly expressed in the lung and the heart [119,166] and, together with ADAMTS-1, -4 and -5, has been detected within human carotid lesions and advanced coronary atherosclerotic plaques [106,107]. ADAMTS-8 expression was increased in the lungs of patients with pulmonary arterial hypertension (PAH) and in mouse/rat models of PAH [119]. In a model of hypoxia-induced PAH, mice bearing a targeted deletion of Adamts8 in pulmonary arterial smooth muscle cells ( Adamts8 ΔSM22α ) showed decreased right ventricular systolic pressure and right ventricular hypertrophy compared with wild-type mice, suggesting a crucial role for ADAMTS-8 in the development of PAH [119].…”