FOXO1 regulates RUNX2 ubiquitination through SMURF2 in calcific aortic valve disease.

The prevalence of calcific aortic valve disease (CAVD) remains substantial while there is currently no medical therapy available. Forkhead box O1 (FOXO1) is known to be involved in the pathogenesis of cardiovascular diseases, including vascular calcification and atherosclerosis; however, its specific role in calcific aortic valve disease remains to be elucidated. In this study, we identified FOXO1 significantly down-regulated in the aortic valve interstitial cells (VICs) of calcified aortic valves by investigating clinical specimens and GEO database analysis. FOXO1 silencing or inhibition promoted VICs osteogenic differentiation in vitro and aortic valve calcification in Apoe-/- mice, respectively. We identified that FOXO1 facilitated the ubiquitination and degradation of RUNX2, which process was mainly mediated by SMAD-specific E3 ubiquitin ligase 2 (SMURF2). Our discoveries unveil a heretofore unacknowledged mechanism involving the FOXO1/SMURF2/RUNX2 axis in CAVD, thereby proposing the potential therapeutic utility of FOXO1 or SMURF2 as viable strategies to impede the progression of CAVD.

Functional Oxidized Hyaluronic Acid Cross-Linked Decellularized Heart Valves for Improved Immunomodulation, Anti-Calcification, and Recellularization.

Tissue engineering heart valves (TEHVs) are expected to address the limitations of mechanical and bioprosthetic valves used in clinical practice. Decellularized heart valve (DHV) is an important scaffold of TEHVs due to its natural three-dimensional structure and bioactive extracellular matrix, but its mechanical properties and hemocompatibility are impaired. In this study, DHV is cross-linked with three different molecular weights of oxidized hyaluronic acid (OHA) by a Schiff base reaction and presented enhanced stability and hemocompatibility, which could be mediated by the molecular weight of OHA. Notably, DHV cross-linked with middle- and high-molecular-weight OHA could drive the macrophage polarization toward the M2 phenotype in vitro. Moreover, DHV cross-linked with middle-molecular-weight OHA scaffolds are further modified with RGD-PHSRN peptide (RPF-OHA/DHV) to block the residual aldehyde groups of the unreacted OHA. The results show that RPF-OHA/DHV not only exhibits anti-calcification properties, but also facilitates endothelial cell adhesion and proliferation in vitro. Furthermore, RPF-OHA/DHV shows excellent performance under an in vivo hemodynamic environment with favorable recellularization and immune regulation without calcification. The optimistic results demonstrate that OHA with different molecular weights has different cross-linking effects on DHV and that RPF-OHA/DHV scaffold with enhanced immune regulation, anti-calcification, and recellularization properties for clinical transformation.

Shear stress activates the Piezo1 channel to facilitate valvular endothelium-oriented differentiation and maturation of human induced pluripotent stem cells.

Valvular endothelial cells (VECs) derived from human induced pluripotent stem cells (hiPSCs) provide an unlimited cell source for tissue engineering heart valves (TEHVs); however, they are limited by their low differentiation efficiency and immature function. In our study, we applied unidirectional shear stress to promote hiPSCs differentiation into valvular endothelial-like cells (VELs). Compared to the static group, shear stress efficiently promoted the differentiation and functional maturation of hiPSC-VELs, as demonstrated by the efficiency of endothelial differentiation reaching 98.3% in the high shear stress group (45 dyn/cm2). Furthermore, we found that Piezo1 served as a crucial mechanosensor for the differentiation and maturation of VELs. Mechanistically, the activation of Piezo1 by shear stress resulted in the influx of calcium ions, which in turn initiated the Akt signaling pathway and promoted the differentiation of hiPSCs into mature VELs. Moreover, VELs cultured on decellularized heart valves (DHVs) exhibited a notable propensity for proliferation, robust adhesion properties, and antithrombotic characteristics, which were dependent on the activation of the Piezo1 channel. Overall, our study demonstrated that proper shear stress activated the Piezo1 channel to facilitate the differentiation and maturation of hiPSC-VELs via the Akt pathway, providing a potential cell source for regenerative medicine, drug screening, pathogenesis, and disease modeling. STATEMENT OF SIGNIFICANCE: This is the first research that systematically analyzes the effect of shear stress on valvular endothelial-like cells (VELs) derived from human induced pluripotent stem cells (hiPSCs). Mechanistically, unidirectional shear stress activates Piezo1, resulting in an elevation of calcium levels, which triggers the Akt signaling pathway and then facilitates the differentiation of functional maturation VELs. After exposure to shear stress, the VELs exhibited enhanced proliferation, robust adhesion capabilities, and antithrombotic characteristics while being cultured on decellularized heart valves. Thus, it is of interest to develop hiPSCs-VELs using shear stress and the Piezo1 channel provides insights into the functional maturation of valvular endothelial cells, thereby serving as a catalyst for potential applications in the development of therapeutic and tissue-engineered heart valves in the future.

Galactose-modified nanoparticles for delivery of microRNA to mitigate the progress of abdominal aortic aneurysms via regulating macrophage polarization.

BACKGROUND: Abdominal aortic aneurysms (AAA) are chronic inflammation in nature and are closely related to macrophages. The purpose was to explore regulating macrophage polarization with target-macrophage nanoparticles impacting the development of AAA. METHODS: Galactose-modified nanoparticles were prepared by self-assembly technology for delivering microRNA (miR)-223. In AngiotensinII-induced experimental AAA model, miR-223-loaded nanoparticles (MirNPs) or PBS was injected at day 7 before and after operation, respectively. Cultured cells and aortic specimen were collected to be analyzed with histology and biochemical examination. RESULTS: In vitro, miR-223 promoted bone marrow-derived macrophages (BMDMs) to polarize to M2. In experimental AAA model, MirNPs significantly decreased the AAA incidence and the ratio of M1 macrophages and production of related proinflammatory cytokines. Furthermore, MirNPs also reduced the expression of the NLRP3 inflammasome. CONCLUSION: Our findings suggested that miR-223-loaded nanoparticles targeting macrophage polarization may mitigate AAA progression via downregulating of NLRP3.

[Progress in the Role of Mechanical Stimulus in Cardiac Development].

Mechanical stimulus is critical to cardiovascular development during embryogenesis period.The mechanoreceptors of endocardial cells and cardiac myocytes may sense mechanical signals and initiate signal transduction that induce gene expression at a cellular level,and then translate molecular-level events into tissue-level deformations,thus guiding embryo development.This review summarizes the regulatory roles of mechanical signals in the early cardiac development including the formation of heart tube,looping,valve and septal morphogenesis,ventricular development and maturation.Further,we discuss the potential mechanical transduction mechanisms of platelet endothelial cell adhesion molecule 1-vascular endothelial-cadherin-vascular endothelial growth factor receptor 2 complex,primary cilia,ion channels,and other mechanical sensors that affect some cardiac malformations.

Application of decellularized vascular matrix in small-diameter vascular grafts.

Coronary artery bypass grafting (CABG) remains the most common procedure used in cardiovascular surgery for the treatment of severe coronary atherosclerotic heart disease. In coronary artery bypass grafting, small-diameter vascular grafts can potentially replace the vessels of the patient. The complete retention of the extracellular matrix, superior biocompatibility, and non-immunogenicity of the decellularized vascular matrix are unique advantages of small-diameter tissue-engineered vascular grafts. However, after vascular implantation, the decellularized vascular matrix is also subject to thrombosis and neoplastic endothelial hyperplasia, the two major problems that hinder its clinical application. The keys to improving the long-term patency of the decellularized matrix as vascular grafts include facilitating early endothelialization and avoiding intravascular thrombosis. This review article sequentially introduces six aspects of the decellularized vascular matrix as follows: design criteria of vascular grafts, components of the decellularized vascular matrix, the changing sources of the decellularized vascular matrix, the advantages and shortcomings of decellularization technologies, modification methods and the commercialization progress as well as the application prospects in small-diameter vascular grafts.

A functional chromogen gene C from wild rice is involved in a different anthocyanin biosynthesis pathway in indica and japonica.

KEY MESSAGE: we identified a functional chromogen gene C from wild rice, providing a new insight of anthocyanin biosynthesis pathway in indica and japonica. Accumulation of anthocyanin is a desirable trait to be selected in rice domestication, but the molecular mechanism of anthocyanin biosynthesis in rice remains largely unknown. In this study, a novel allele of chromogen gene C, OrC1, from Oryza rufipongon was cloned and identified as a determinant regulator of anthocyanin biosynthesis. Although OrC1 functions in purple apiculus, leaf sheath and stigma in indica background, it only promotes purple apiculus in japonica. Transcriptome analysis revealed that OrC1 regulates flavonoid biosynthesis pathway and activates a few bHLH and WD40 genes of ternary MYB-bHLH-WD40 complex in indica. Differentially expressed genes and metabolites were found in the indica and japonica backgrounds, indicating that OrC1 activated the anthocyanin biosynthetic genes OsCHI, OsF3H and OsANS and produced six metabolites independently. Artificial selection and domestication of C1 gene in rice occurred on the coding region in the two subspecies independently. Our results reveal the regulatory system and domestication of C1, provide new insights into MYB transcript factor involved in anthocyanin biosynthesis, and show the potential of engineering anthocyanin biosynthesis in rice.

Which is the best prosthesis in an isolated or combined tricuspid valve replacement?

OBJECTIVES: The debate concerning the optimal choice of tricuspid position continues. We compared the long-term results of mechanical and biological prostheses in patients who underwent isolated or combined tricuspid valve replacement, at 2 major cardiac surgical centres in central China. METHODS: From January 1999 to December 2018, 338 patients underwent tricuspid valve replacement. Patients were divided into an isolated group or a combined group according to whether their surgery was combined with a left heart valve surgery. Mechanical tricuspid valve replacement was performed in 142 patients (isolated group: 41 vs combined group: 101), and 196 patients underwent bioprosthetic tricuspid valve replacement (isolated group: 145 vs combined group: 51). Operative results, long-term survival and tricuspid valve-related events were compared. RESULTS: Early mortality in the combined group was higher (n = 6, 4%) than that in the isolated group (n = 3, 2%), but no significant difference was observed between the mechanical and biological subgroups. In the isolated group, there was a higher event-free rate in the biological subgroup than in the mechanical subgroup (P = 0.042) and a similar result was also observed for patients without Ebstein's anomaly (P = 0.039). In the combined group, no significant difference was observed (P = 0.98). Survival rates were similar between the mechanical and biological subgroups in both the isolated (P = 0.54) and combined (P = 0.81) groups. Mechanical valves in isolated tricuspid valve replacement were more prone to valve thrombosis and bleeding. CONCLUSIONS: Every decision regarding tricuspid valve prostheses should be individualized, but biological prostheses may be an optimal choice for patients, especially for patients without Ebstein's anomaly, in isolated tricuspid valve replacement.

Real time three-dimensional echocardiographic quantification of left atrial volume in orthotopic heart transplant recipients: Comparisons with cardiac magnetic resonance imaging.

INTRODUCTION: The accuracy of real time three-dimensional echocardiography (RT-3DE) in evaluating left atrial volume (LAV) of heart transplant recipients against cardiac magnetic resonance (CMR) has not been reported. The aim of this study was to compare LAV with RT-3DE with respect to CMR in heart transplant recipients. METHODS: Thirty-one heart transplant recipients who received echocardiogram and CMR examination on the same day were prospectively enrolled. The maximal LAV, minimal LAV by RT-3DE, and two-dimensional echocardiography (2DE) were compared with CMR measurements. Inter-technique comparisons included Pearson's correlation coefficient and Bland-Altman analysis. Reproducibility of 2DE and RT-3DE technique was assessed by intra-class correlation coefficient (ICC). RESULTS: RT-3DE-derived LAV values showed higher correlation with CMR than 2DE measurements in heart transplant recipients (r = .93 vs r = .76 for maximal LAV; r = .91 vs r = .81 for minimal LAV). Two-dimensional echocardiography underestimated maximal LAV by 10 ± 31 mL and minimal LAV by 26 ± 26 mL. Although RT-3DE underestimated minimal LAV 15 ± 19 mL, no significant difference between RT-3DE and CMR was observed in maximal LAV (RT-3DE: 86 ± 22 mL; CMR: 89 ± 23 mL, P = .079), with a negligible bias of 3 mL. Inter-observer and intra-observer agreement were excellent for 2DE and RT-3DE parameters. CONCLUSION: Compared with CMR reference, RT-3DE-derived LAV measurements are more accurate than 2DE-based analysis in heart transplant recipients, especially with regard to the assessment of maximal LAV. RT-3DE may be a valid alternative to CMR for quantification LAV in heart transplant recipients.

Modifying decellularized aortic valve scaffolds with stromal cell-derived factor-1α loaded proteolytically degradable hydrogel for recellularization and remodeling.

Decellularized matrix is of great interest as a scaffold for the tissue engineering heart valves due to its naturally three-dimensional structure and bioactive composition. A primary challenge of tissue engineered heart valves based on decellularized matrix is to grow a physiologically appropriate cell population within the leaflet tissue. In this study, a composite scaffold was fabricated by the combination of a porous matrix metalloproteinase (MMP) degradable poly (ethylene glycol) (PEG) hydrogel that were loaded with stromal cell-derived factor-1α (SDF-1α) and a mechanically supportive decellularized porcine aortic valve. Results demonstrated that the modified scaffold enhanced bone marrow mesenchymal stem cells (BMSC) adhesion, viability and proliferation, and promoted BMSC differentiate into valve interstitial-like cells. Furthermore, these modifications lead to enhanced protection of the scaffold from thrombosis. In vivo assessment by rat subdermal model showed the modified scaffold was highly biocompatible with tissue remodeling characterized by promoting mesenchymal stem cells recruitment and facilitating M2 macrophage phenotype polarization. The surface layers of PEG hydrogel not only could provide a niche for cell migration, proliferation and differentiation, but also protect the scaffolds from rapid degeneration, inflammation and calcification. The intermediate layer of decellularized valve could maintain the organization of the scaffold and perform the valve function. The promising results emphasize the potential of our scaffolds to improve recellularization and promote remodeling of implanted decellularized valves. These findings suggest that the SDF-1α loaded MMP degradable PEG hydrogel modification could be an efficient approach to develop functional decellularized heart valve. STATEMENT OF SIGNIFICANCE: A composite scaffold was fabricated by the combination of a porous matrix metalloproteinase (MMP) degradable poly (ethylene glycol) (PEG) hydrogel that were loaded with SDF-1α and a mechanically supportive decellularized porcine aortic valve. The surface layers of PEG hydrogel not only could provide a niche for cell migration, proliferation and differentiation, but also protect the scaffolds from rapid degeneration, inflammation and calcification. The intermediate layer of decellularized valve could maintain the organization of the scaffold and perform the valve function. The promising results emphasize the ability of our scaffolds to improve recellularization and promote remodeling of implanted decellularized valves. This suggests that the extracellular matrix-based valve scaffolds have potential for clinical applications.

The shift of macrophages toward M1 phenotype promotes aortic valvular calcification.

OBJECTIVE: The purpose of the present study was to comprehensively compare the phenotype profile of infiltrated macrophages in human noncalcified and calcific aortic valves, and to determine whether the shift of macrophage polarization modulates valvular calcification in vitro. METHODS: Cell surface markers of macrophages and inflammatory cytokines expression in 90 cases of human noncalcified and calcific aortic valve leaflets were analyzed. The normal aortic valve interstitial cells were isolated and cultured in vitro. After incubation with nonconditioned medium and conditioned medium from unstimulated or lipopolysaccharide-stimulated U937 monocytes, valve interstitial cells were evaluated by osteogenic differentiation markers. RESULTS: Infiltration of macrophages was enhanced in the calcific aortic valves, and M1 phenotype was the predominant macrophage subsets. In addition, both proinflammatory and anti-inflammatory cytokines were significantly upregulated in the calcific aortic valves. Furthermore, lipopolysaccharide-stimulated monocytes presented with increased expression of inducible nitric oxide synthase and high proportional CD11c-positive (M1) macrophages. Conditioned medium from unstimulated monocytes promoted the osteogenic differentiation of valve interstitial cells in vitro, as evidenced by increased markers such as bone morphogenetic protein 2, osteopontin, and alkaline phosphatase. Conditioned medium from M1 macrophages further enhanced valve interstitial cells calcification. Enzyme-linked immunosorbent assay showed that M1 phenotype macrophages secreted tumor necrosis factors α and interleukin 6, and neutralizing antibodies to these 2 proinflammatory cytokines attenuated induction of osteogenic differentiation and calcification by the conditioned media. CONCLUSIONS: Both total numbers and polarization of macrophage influence the process of calcification in human aortic valve. The shift toward M1 phenotype might promote valve interstitial cell calcification.

Should a Mechanical or Biological Prosthesis Be Used for a Tricuspid Valve Replacement? A Meta-Analysis.

BACKGROUND AND AIM OF THE STUDY: The prosthesis of choice for a tricuspid valve replacement is still unkown. This meta-analysis was undertaken to review the results of mechanical and bioprosthetic valves in the tricuspid position. METHODS: We identified all relevant studies published in the past 20 years (from January 1, 1995 to December 31, 2014) through the Embase, Current Contents, and PubMed databases. The hazard ratio and its 95% confidence limits were utilized to evaluate time-to-event related effects of surgical procedures. The Q-statistic, Index of Inconsistency test, funnel plots, and Egger's test were used to assess the degree of heterogeneity and publication bias. Random effects models were used, and study quality was also assessed. RESULTS: In our meta-analysis, 22 studies published from 1995 to 2014 were reviewed and 2630 patients and 14,694 follow-up years were analyzed. No statistically significant difference was identified between mechanical and biological valves in terms of survival, reoperation, and prosthetic valve failure. The respective pooled hazard ratio estimates were 0.95 (0.79 to 1.16, p = 0.62, I(2) = 29%), 1.20 (0.84 to 1.71, p = 0.33, I(2) = 0%), and 0.35 (0.06 to 2.01, p = 0.24, I(2) = 0%). A higher risk of thrombosis was found in mechanical tricuspid valve prostheses (3.86, 1.38 to 10.82, p = 0.01, I(2) = 0%). CONCLUSIONS: No statistically significant difference was identified between mechanical and biological valves in terms of survival, reoperation, or prosthetic valve failure, but mechanical tricuspid valve prostheses had a higher risk of thrombosis. doi: 10.1111/jocs.12730 (J Card Surg 2016;31:294-302).

Domestication and geographic origin of Oryza sativa in China: insights from multilocus analysis of nucleotide variation of O. sativa and O. rufipogon.

Previous studies have indicated that China is one of the domestication centres of Asian cultivated rice (Oryza sativa), and common wild rice (O. rufipogon) is the progenitor of O. sativa. However, the number of domestication times and the geographic origin of Asian cultivated rice in China are still under debate. In this study, 100 accessions of Asian cultivated rice and 111 accessions of common wild rice in China were selected to examine the relationship between O. sativa and O. rufipogon and thereby infer the domestication and evolution of O. sativa in China through sequence analyses of six gene regions, trnC-ycf6 in chloroplast genomes, cox3 in mitochondrial genomes and ITS, Ehd1, Waxy, Hd1 in nuclear genomes. The results indicated that the two subspecies of O. sativa (indica and japonica) were domesticated independently from different populations of O. rufipogon with gene flow occurring later from japonica to indica; Southern China was the genetic diversity centre of O. rufipogon, and the Pearl River basin near the Tropic of Cancer was the domestication centre of O. sativa in China.