scholarly journals Phosphoproteomic Analysis Reveals Downstream PKA Effectors of AKAP Cypher/ZASP in the Pathogenesis of Dilated Cardiomyopathy

2021 ◽  
Vol 8 ◽  
Author(s):  
Jialan Lv ◽  
Zhicheng Pan ◽  
Jian Chen ◽  
Rui Xu ◽  
Dongfei Wang ◽  
...  
Keyword(s):  
Dilated Cardiomyopathy ◽  
Knockout Mice ◽  
Cardiac Tissue ◽  
Troponin I ◽  
Bioinformatic Analysis ◽  
Differentially Expressed ◽  
Cardiomyocyte Proliferation ◽  
Intercalated Discs ◽  
Anchoring Protein ◽  
Alternatively Spliced

Background: Dilated cardiomyopathy (DCM) is a major cause of heart failure worldwide. The Z-line protein Cypher/Z-band alternatively spliced PDZ-motif protein (ZASP) is closely associated with DCM, both clinically and in animal models. Our earlier work revealed Cypher/ZASP as a PKA-anchoring protein (AKAP) that tethers PKA to phosphorylate target substrates. However, the downstream PKA effectors regulated by AKAP Cypher/ZASP and their relevance to DCM remain largely unknown.Methods and Results: For the identification of candidate PKA substrates, global quantitative phosphoproteomics was performed on cardiac tissue from wild-type and Cypher-knockout mice with PKA activation. A total of 216 phosphopeptides were differentially expressed in the Cypher-knockout mice; 31 phosphorylation sites were selected as candidates using the PKA consensus motifs. Bioinformatic analysis indicated that differentially expressed proteins were enriched mostly in cell adhesion and mRNA processing. Furthermore, the phosphorylation of β-catenin Ser675 was verified to be facilitated by Cypher. This phosphorylation promoted the transcriptional activity of β-catenin, and also the proliferative capacity of cardiomyocytes. Immunofluorescence staining demonstrated that Cypher colocalised with β-catenin in the intercalated discs (ICD) and altered the cytoplasmic distribution of β-catenin. Moreover, the phosphorylation of two other PKA substrates, vimentin Ser72 and troponin I Ser23/24, was suppressed by Cypher deletion.Conclusions: Cypher/ZASP plays an essential role in β-catenin activation via Ser675 phosphorylation, which modulates cardiomyocyte proliferation. Additionally, Cypher/ZASP regulates other PKA effectors, such as vimentin Ser72 and troponin I Ser23/24. These findings establish the AKAP Cypher/ZASP as a signalling hub in the progression of DCM.

Abstract 1066: Cardiac-restricted Disruption of Supressor of Cytokine Signaling-3 Leads to Dilated Cardiomyopathy in a gp130 Independent Manner

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Toshitaka Yajima ◽  
Hideo Yasukawa ◽  
Hanbing Zhou ◽  
Tomoko Yajima ◽  
Byung-Kwan Lim ◽  
...  
Keyword(s):  
Dilated Cardiomyopathy ◽  
Knockout Mice ◽  
Cardiac Myocyte ◽  
Physiological Role ◽  
Calcium Sensitivity ◽  
Cytokine Signaling ◽  
Independent Manner ◽  
Downstream Signaling ◽  
Intercalated Discs

We recently reported that cardiac-restricted overexpression of suppressor of cytokine signaling-3 (SOCS3) markedly increases cardiac myocyte susceptibility to virus infection. This suggests that endogenous SOCS3 also has significant biological effect in the cardiac myocyte. To understand physiological role of SOCS3 in the cardiac myocyte, we generated cardiac-specific SOCS3 knockout mice. The mice were born at the expected Mendelian ratio, but developed cardiac dysfunction (%FS; wt: 45±2.6 vs. ko: 31±4.5, mean±SE, n=8, p=0.017, 5-month-old) estimated by echocardiogram from 5 months of age and died with signs of dyspnea by 7 months of age. Postmortem analysis revealed significant pleural effusions and ascites, consistent with the presence of heart failure. Histological analysis showed that the heart has a thin left and right ventricular walls with chamber dilatation; however, typical findings of a myopathic heart such as myocyte disarray, inflammation and fibrosis were only rarely observed. Ultrastructure analysis of the dilated heart samples showed intact myofibrils, mitochondria, intercalated discs and gap junctions, suggesting that the myocyte had non-structural abnormalities that led to functional impairment. Unexpectedly calcium transients in the isolated adult myocytes from the failing hearts were significantly increased as compared to those from age-matched normal hearts (Fura-2 intensity ratio 340/380 nm; wt: 0.44±0.01 vs. ko: 0.54±0.01, mean±SE, n=3, p<0.0002), suggesting the decreased calcium sensitivity of SOCS3 deficient myocytes. We also observed spontaneous ventricular tachycardia in the knockout mice by telemetry analysis. Since SOCS3 has been reported to be a crucial negative-feedback regulator of gp130 signaling in vivo, we sought to determine whether the phenotype is due to overactivation of cardiac gp130 signaling. While gp130 downstream signaling such as STAT3, ERK and AKT pathways were significantly activated in the knockout heart, cardiac-specific knockout of both gp130 and SOCS3 did not rescue the dilated cardiomyopathy phenotype. Conclusion: These results indicate that a gp130-independent mechanism is responsible for the dilated cardiomyopathy that occurs in SOCS3 deficient hearts.

294 Clinical significance of troponin I efflux and troponin autoantibodies in patients with dilated cardiomyopathy

2006 ◽  
Vol 5 (1) ◽  
pp. 68-68
Author(s):  
K MIETTINEN ◽  
S ERIKSSON ◽  
J MAGGA ◽  
P TUOMAINEN ◽  
E VANNINEN ◽  
...  
Keyword(s):  
Dilated Cardiomyopathy ◽  
Clinical Significance ◽  
Troponin I

scholarly journals Sex-Biased lncRNA Signature in Fetal Growth Restriction (FGR)

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 921
Author(s):  
Aleksandra Lipka ◽  
Jan Pawel Jastrzebski ◽  
Lukasz Paukszto ◽  
Karol Gustaw Makowczenko ◽  
Elzbieta Lopienska-Biernat ◽  
...  
Keyword(s):  
Fetal Growth ◽  
Fetal Growth Restriction ◽  
Target Genes ◽  
Active Regions ◽  
Bioinformatic Analysis ◽  
Growth Restriction ◽  
Differentially Expressed ◽  
Circular Rnas ◽  
Differential Analysis ◽  
Protein Coding

Impaired fetal growth is one of the most important causes of prematurity, stillbirth and infant mortality. The pathogenesis of idiopathic fetal growth restriction (FGR) is poorly understood but is thought to be multifactorial and comprise a range of genetic causes. This research aimed to investigate non-coding RNAs (lncRNAs) in the placentas of male and female fetuses affected by FGR. RNA-Seq data were analyzed to detect lncRNAs, their potential target genes and circular RNAs (circRNAs); a differential analysis was also performed. The multilevel bioinformatic analysis enabled the detection of 23,137 placental lncRNAs and 4263 of them were classified as novel. In FGR-affected female fetuses’ placentas (ff-FGR), among 19 transcriptionally active regions (TARs), five differentially expressed lncRNAs (DELs) and 12 differentially expressed protein-coding genes (DEGs) were identified. Within 232 differentially expressed TARs identified in male fetuses (mf-FGR), 33 encompassed novel and 176 known lncRNAs, and 52 DEGs were upregulated, while 180 revealed decreased expression. In ff-FGR ACTA2-AS1, lncRNA expression was significantly correlated with five DEGs, and in mf-FGR, 25 TARs were associated with DELs correlated with 157 unique DEGs. Backsplicing circRNA processes were detected in the range of H19 lncRNA, in both ff- and mf-FGR placentas. The performed global lncRNAs characteristics in terms of fetal sex showed dysregulation of DELs, DEGs and circRNAs that may affect fetus growth and pregnancy outcomes. In female placentas, DELs and DEGs were associated mainly with the vasculature, while in male placentas, disturbed expression predominantly affected immune processes.

Application potential of three-dimensional silk fibroin scaffold using mesenchymal stem cells for cardiac regeneration

2021 ◽  
pp. 088532822110185
Author(s):  
Yuksel Cetin ◽  
Merve G Sahin ◽  
Fatma N Kok
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Silk Fibroin ◽  
Cardiac Tissue ◽  
Troponin I ◽  
Cardiac Tissue Engineering ◽  
Swelling Ratio ◽  
Differentiation Potential ◽  
Cardiomyogenic Differentiation

Cardiac tissue engineering focusing on biomaterial scaffolds incorporating cells from different sources has been explored to regenerate or repair damaged area as a lifesaving approach.The aim of this study was to evaluate the cardiomyocyte differentiation potential of human adipose mesenchymal stem cells (hAD-MSCs) as an alternative cell source on silk fibroin (SF) scaffolds for cardiac tissue engineering. The change in surface morphology of SF scaffolds depending on SF concentration (1–6%, w/v) and increase in their porosity upon application of unidirectional freezing were visualized by scanning electron microscopy (SEM). Swelling ratio was found to increase 2.4 fold when SF amount was decreased from 4% to 2%. To avoid excessive swelling, 4% SF scaffold with swelling ratio of 10% (w/w) was chosen for further studies.Biodegradation rate of SF scaffolds depended on enzymatic activity was found to be 75% weight loss of SF scaffolds at the day 14. The phenotype of hAD-MSCs and their multi-linage potential into chondrocytes, osteocytes, and adipocytes were shown by flow cytometry and immunohistochemical staining, respectively.The viability of hAD-MSCs on 3D SF scaffolds was determined as 90%, 118%, and 138% after 1, 7, and 14 days, respectively. The use of 3D SF scaffolds was associated with increased production of cardiomyogenic biomarkers: α-actinin, troponin I, connexin 43, and myosin heavy chain. The fabricated 3D SF scaffolds were proved to sustain hAD-MSCs proliferation and cardiomyogenic differentiation therefore, hAD-MSCs on 3D SF scaffolds may useful tool to regenerate or repair damaged area using cardiac tissue engineering techniques.

scholarly journals Polyphenol Supplementation Reverses Age-Related Changes in Microglial Signaling Cascades

2021 ◽  
Vol 22 (12) ◽  
pp. 6373
Author(s):  
Ahmad Jalloh ◽  
Antwoine Flowers ◽  
Charles Hudson ◽  
Dale Chaput ◽  
Jennifer Guergues ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Control Diet ◽  
Bioinformatic Analysis ◽  
Diet Group ◽  
Differentially Expressed ◽  
Signaling Cascades ◽  
Complex Nature ◽  
Differentially Expressed Proteins ◽  
Ms Analysis ◽  
Age Related

Microglial activity in the aging neuroimmune system is a central player in aging-related dysfunction. Aging alters microglial function via shifts in protein signaling cascades. These shifts can propagate neurodegenerative pathology. Therapeutics require a multifaceted approach to understand and address the stochastic nature of this process. Polyphenols offer one such means of rectifying age-related decline. Our group used mass spectrometry (MS) analysis to explicate the complex nature of these aging microglial pathways. In our first experiment, we compared primary microglia isolated from young and aged rats and identified 197 significantly differentially expressed proteins between these groups. Then, we performed bioinformatic analysis to explore differences in canonical signaling cascades related to microglial homeostasis and function with age. In a second experiment, we investigated changes to these pathways in aged animals after 30-day dietary supplementation with NT-020, which is a blend of polyphenols. We identified 144 differentially expressed proteins between the NT-020 group and the control diet group via MS analysis. Bioinformatic analysis predicted an NT-020 driven reversal in the upregulation of age-related canonical pathways that control inflammation, cellular metabolism, and proteostasis. Our results highlight salient aspects of microglial aging at the level of protein interactions and demonstrate a potential role of polyphenols as therapeutics for age-associated dysfunction.

scholarly journals Disorganization of intercalated discs in dilated cardiomyopathy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yukinobu Ito ◽  
Makoto Yoshida ◽  
Hirotake Masuda ◽  
Daichi Maeda ◽  
Yukitsugu Kudo-Asabe ◽  
...  
Keyword(s):  
Dilated Cardiomyopathy ◽  
Interstitial Fibrosis ◽  
Left Ventricular ◽  
Pathological Diagnosis ◽  
Control Group ◽  
Failure Group ◽  
Intercalated Discs ◽  
Group Reduction ◽  
Group A ◽  
Morphological Differences

AbstractDilated cardiomyopathy (DCM) is a primary myocardial disease, the pathology of which is left ventricular or biventricular dilation and impaired myocardial contractility. The clinical and pathological diagnosis of DCM is difficult, and other cardiac diseases must be ruled out. Several studies have reported pathological findings that are characteristic of DCM, including cardiomyocyte atrophy, nuclear pleomorphism, and interstitial fibrosis, but none of these findings are DCM-specific. In this study, we examined the morphological differences in the intercalated discs (ICDs) between three groups of patients, a DCM group, a chronic heart failure group, and a control group. A total of 22 autopsy cases, including five DCM cases, nine CHF cases and eight control cases, were retrieved from the archives of the Department of Pathology at Akita University, Japan. The morphological differences were examined using multiple methods: macroscopic examination, light microscopy, immunohistochemistry, electron microscopy, and gene expression analyses. We observed disorganized ICDs, clearly illustrated by N-cadherin immunostaining in the DCM group. “Reduction of N-cadherin immunostaining intensity” and “ICD scattering” was DCM-specific. The results suggest that disorganized ICDs contribute to the development of DCM, and that N-cadherin immunostaining is useful for determining the presence of disorganized ICDs and for the pathological diagnosis of DCM.

Novel mutation in cardiac troponin I in recessive idiopathic dilated cardiomyopathy

The Lancet ◽  
2004 ◽  
Vol 363 (9406) ◽  
pp. 371-372 ◽  
Author(s):  
Ross T Murphy ◽  
Jens Mogensen ◽  
Anthony Shaw ◽  
Toru Kubo ◽  
Sian Hughes ◽  
...  
Keyword(s):  
Dilated Cardiomyopathy ◽  
Cardiac Troponin ◽  
Troponin I ◽  
Novel Mutation ◽  
Cardiac Troponin I ◽  
Idiopathic Dilated Cardiomyopathy

scholarly journals Defects in caveolin-1 cause dilated cardiomyopathy and pulmonary hypertension in knockout mice

2002 ◽  
Vol 99 (17) ◽  
pp. 11375-11380 ◽  
Author(s):  
Y.-Y. Zhao ◽  
Y. Liu ◽  
R.-V. Stan ◽  
L. Fan ◽  
Y. Gu ◽  
...  
Keyword(s):  
Pulmonary Hypertension ◽  
Dilated Cardiomyopathy ◽  
Knockout Mice ◽  
Caveolin 1

PRMT5 Prevents Dilated Cardiomyopathy via Suppression of Protein O-GlcNAcylation

2021 ◽  
Author(s):  
Zhenhua Li ◽  
Jingping Xu ◽  
Yao Song ◽  
Chong Xin ◽  
Lantao Liu ◽  
...  
Keyword(s):  
Dilated Cardiomyopathy ◽  
Knockout Mice ◽  
Rna Splicing ◽  
Gene Knockout ◽  
Major Type ◽  
Integrated Analysis ◽  
Arginine Methyltransferase ◽  
Potential Strategy ◽  
Glcnac Transferase

Rationale: Protein O-GlcNAcylation is dynamically regulated by two key enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Excessive protein O-GlcNAcylation contributes to dilated cardiomyopathy (DCM), but its regulatory mechanisms are not fully understood. The protein arginine methyltransferase 5 (PRMT5) is the major type II arginine methyltransferase, which plays critical physiological roles by symmetrically dimethylating various downstream targets including proteins involved in RNA splicing. However, its function in regulating protein O-GlcNAcylation and DCM is unexplored. Objective: To elucidate the physiological function of PRMT5 and the mechanism underlying its role in regulating cardiac O-GlcNAcylation and homeostasis. Methods and Results: Conditional gene knockout was used to study the in vivo function of Prmt5 in regulating cardiac homeostasis. An integrated analysis of transcriptomic and metabolomic profiles was performed to investigate the molecular mechanism. Adeno-associated virus 9 (AAV9)-mediated gene delivery in the mouse was used to study the protein O-GlcNAcylation in Prmt5 deficiency-induced DCM. PRMT5 mRNA was decreased in human DCM hearts, and cardiomyocyte-specific Prmt5 deletion in mice resulted in DCM and heart failure. Transcriptomic and metabolomic profiling identified increased O-GlcNAcylation in the hearts of Prmt5-knockout mice. Mechanistically, Prmt5 deletion suppressed O-GlcNAcase (OGA) expression by inhibiting the transcription of Oga and triggering its aberrant splicing. Consistently, a positive correlation of PRMT5 and OGA was identified in human DCM hearts. Notably, gene therapy with AAV9 encoding the correctly spliced Oga normalized the cardiac protein O-GlcNAcylation levels and partially rescued the dilation and dysfunction of the hearts in Prmt5-knockout mice. Conclusions: Our data demonstrate a novel function of PRMT5 in regulating protein O-GlcNAcylation to maintain cardiac homeostasis, suggesting that targeting the PRMT5-OGA axis could be a potential strategy for treating DCM.

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