ABSTRACT
BACKGROUND: An imbalance of antiproliferative BMP (bone morphogenetic protein) signaling and proliferative TGF-ß (transforming growth factor-ß) signaling is implicated in the development of pulmonary arterial hypertension (PAH). The posttranslational modification (eg, phosphorylation and ubiquitination) of TGF-ß family receptors, including BMPR2 (bone morphogenetic protein type 2 receptor)/ALK2 (activin receptor-like kinase-2) and TGF-ßR2/R1, and receptor-regulated Smads significantly affects their activity and thus regulates the target cell fate. BRCC3 modifies the activity and stability of its substrate proteins through K63-dependent deubiquitination. By modulating the posttranslational modifications of the BMP/TGF-ß-PPARγ pathway, BRCC3 may play a role in pulmonary vascular remodeling, hence the pathogenesis of PAH. METHODS: Bioinformatic analyses were used to explore the mechanism by which BRCC3 deubiquitinates ALK2. Cultured pulmonary artery smooth muscle cells (PASMCs), mouse models, and specimens from patients with idiopathic PAH were used to investigate the rebalance between BMP and TGF-ß signaling in regulating ALK2 phosphorylation and ubiquitination in the context of pulmonary hypertension. RESULTS: BRCC3 was significantly downregulated in PASMCs from patients with PAH and animals with experimental pulmonary hypertension. BRCC3, by de-ubiquitinating ALK2 at Lys-472 and Lys-475, activated receptor-regulated Smad1/5/9, which resulted in transcriptional activation of BMP-regulated PPARγ, p53, and Id1. Overexpression of BRCC3 also attenuated TGF-ß signaling by downregulating TGF-ß expression and inhibiting phosphorylation of Smad3. Experiments in vitro indicated that overexpression of BRCC3 or the de-ubiquitin-mimetic ALK2-K472/475R attenuated PASMC proliferation and migration and enhanced PASMC apoptosis. In SM22α-BRCC3-Tg mice, pulmonary hypertension was ameliorated because of activation of the ALK2-Smad1/5-PPARγ axis in PASMCs. In contrast, Brcc3-/- mice showed increased susceptibility of experimental pulmonary hypertension because of inhibition of the ALK2-Smad1/5 signaling. CONCLUSIONS: These results suggest a pivotal role of BRCC3 in sustaining pulmonary vascular homeostasis by maintaining the integrity of the BMP signaling (ie, the ALK2-Smad1/5-PPARγ axis) while suppressing TGF-ß signaling in PASMCs. Such rebalance of BMP/TGF-ß pathways is translationally important for PAH alleviation.
Subject(s)
Hypertension, Pulmonary , Muscle, Smooth, Vascular , Myocytes, Smooth Muscle , Animals , Humans , Male , Mice , Activin Receptors, Type II/metabolism , Activin Receptors, Type II/genetics , Bone Morphogenetic Protein Receptors, Type II/metabolism , Bone Morphogenetic Protein Receptors, Type II/genetics , Cell Proliferation , Cells, Cultured , Disease Models, Animal , Hypertension, Pulmonary/metabolism , Hypertension, Pulmonary/genetics , Hypertension, Pulmonary/pathology , Mice, Inbred C57BL , Mice, Knockout , Muscle, Smooth, Vascular/metabolism , Muscle, Smooth, Vascular/pathology , Myocytes, Smooth Muscle/metabolism , Myocytes, Smooth Muscle/pathology , PPAR gamma/metabolism , PPAR gamma/genetics , Pulmonary Arterial Hypertension/metabolism , Pulmonary Arterial Hypertension/pathology , Pulmonary Arterial Hypertension/genetics , Pulmonary Artery/metabolism , Pulmonary Artery/pathology , Signal Transduction , Ubiquitination , Vascular RemodelingABSTRACT
PURPOSE: Glycolysis and immune metabolism play important roles in acute myocardial infarction (AMI). Therefore, this study aimed to identify and experimentally validate the glycolysis-related hub genes in AMI as diagnostic biomarkers, and further explore the association between hub genes and immune infiltration. METHODS: Differentially expressed genes (DEGs) from AMI peripheral blood mononuclear cells (PBMCs) were analyzed using R software. Glycolysis-related DEGs (GRDEGs) were identified and analyzed using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) for functional enrichment. A protein-protein interaction network was constructed using the STRING database and visualized using Cytoscape software. Immune infiltration analysis between patients with AMI and stable coronary artery disease (SCAD) controls was performed using CIBERSORT, and correlation analysis between GRDEGs and immune cell infiltration was performed. We also plotted nomograms and receiver operating characteristic (ROC) curves to assess the predictive accuracy of GRDEGs for AMI occurrence. Finally, key genes were experimentally validated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting using PBMCs. RESULTS: A total of 132 GRDEGs and 56 GRDEGs were identified on the first day and 4-6 days after AMI, respectively. Enrichment analysis indicated that these GRDEGs were mainly clustered in the glycolysis/gluconeogenesis and metabolic pathways. Five hub genes (HK2, PFKL, PKM, G6PD, and ALDOA) were selected using the cytoHubba plugin. The link between immune cells and hub genes indicated that HK2, PFKL, PKM, and ALDOA were significantly positively correlated with monocytes and neutrophils, whereas G6PD was significantly positively correlated with neutrophils. The calibration curve, decision curve analysis, and ROC curves indicated that the five hub GRDEGs exhibited high predictive value for AMI. Furthermore, the five hub GRDEGs were validated by RT-qPCR and western blotting. CONCLUSION: We concluded that HK2, PFKL, PKM, G6PD, and ALDOA are hub GRDEGs in AMI and play important roles in AMI progression. This study provides a novel potential immunotherapeutic method for the treatment of AMI.
Subject(s)
Computational Biology , Gene Regulatory Networks , Glycolysis , Myocardial Infarction , Protein Interaction Maps , Humans , Glycolysis/genetics , Myocardial Infarction/genetics , Myocardial Infarction/immunology , Myocardial Infarction/diagnosis , Gene Expression Profiling , Databases, Genetic , Transcriptome , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/metabolism , Predictive Value of Tests , Male , Middle Aged , Hexokinase/genetics , Female , Case-Control Studies , Nomograms , Reproducibility of ResultsABSTRACT
All-solid-state high-performance asymmetric supercapacitors (ASCs) are fabricated using γ-MnS as positive electrode and porous eggplant derived activated carbon (EDAC) as negative electrode with saturated potassium hydroxide agar gel as the solid electrolyte. The laminar wurtzite nanostructure of γ-MnS facilitates the insertion of hydroxyl ions into the interlayer space, and the manganese sulfide nanowire offers electronic transportation channels. The size-uniform porous nanostructure of EDAC provides a continuous electron pathway as well as facilitates short ionic transportation pathways. Due to these special nanostructures of both the MnS and the EDAC, they exhibited a specific capacitance of 573.9 and 396 F g(-1) at 0.5 A g(-1), respectively. The optimized MnS//EDAC asymmetric supercapacitor shows a superior performance with specific capacitance of 110.4 F g(-1) and 89.87% capacitance retention after 5000 cycles, a high energy density of 37.6 Wh kg(-1) at a power density of 181.2 W kg(-1) and remains 24.9 Wh kg(-1) even at 5976 W kg(-1). Impressively, such two assembled all-solid-state cells in series can light up a red LED indicator for 15 minutes after fully charged. These impressive results make these pollution-free materials promising for practical applications in solid aqueous electrolyte-based ASCs.