Galectin 3 enhances platelet aggregation and thrombosis via Dectin-1 activation: a translational study.

AIMS: Galectin-3, a ß-galactoside-binding lectin, is abnormally increased in cardiovascular disease. Plasma Galectin-3 receives a Class II recommendation for heart failure management and has been extensively studied for multiple cellular functions. The direct effects of Galectin-3 on platelet activation remain unclear. This study explores the direct effects of Galectin-3 on platelet activation and thrombosis. METHODS AND RESULTS: A strong positive correlation between plasma Galectin-3 concentration and platelet aggregation or whole blood thrombus formation was observed in patients with coronary artery disease (CAD). Multiple platelet function studies demonstrated that Galectin-3 directly potentiated platelet activation and in vivo thrombosis. Mechanistic studies using the Dectin-1 inhibitor, laminarin, and Dectin-1-/- mice revealed that Galectin-3 bound to and activated Dectin-1, a receptor not previously reported in platelets, to phosphorylate spleen tyrosine kinase and thus increased Ca2+ influx, protein kinase C activation, and reactive oxygen species production to regulate platelet hyperreactivity. TD139, a Galectin-3 inhibitor in a Phase II clinical trial, concentration dependently suppressed Galectin-3-potentiated platelet activation and inhibited occlusive thrombosis without exacerbating haemorrhage in ApoE-/- mice, which spontaneously developed increased plasma Galectin-3 levels. TD139 also suppressed microvascular thrombosis to protect the heart from myocardial ischaemia-reperfusion injury in ApoE-/- mice. CONCLUSION: Galectin-3 is a novel positive regulator of platelet hyperreactivity and thrombus formation in CAD. As TD139 has potent antithrombotic effects without bleeding risk, Galectin-3 inhibitors may have therapeutic advantages as potential antiplatelet drugs for patients with high plasma Galectin-3 levels.

Enhanced myofilament calcium sensitivity aggravates abnormal calcium handling and diastolic dysfunction in patient-specific induced pluripotent stem cell-derived cardiomyocytes with MYH7 mutation.

Hypertrophic cardiomyopathy (HCM), the most common inherited heart disease, is frequently caused by mutations in the ß-cardiac myosin heavy chain gene (MYH7). Abnormal calcium handling and diastolic dysfunction are archetypical features of HCM caused by MYH7 gene mutations. However, the mechanism of how MYH7 mutations leads to these features remains unclear, which inhibits the development of effective therapies. Initially, cardiomyocytes were generated from induced pluripotent stem cells from an eight-year-old girl diagnosed with HCM carrying a MYH7(C.1063 G>A) heterozygous mutation(mutant-iPSC-CMs) and mutation-corrected isogenic iPSCs(control-iPSC-CMs) in the present study. Next, we compared phenotype of mutant-iPSC-CMs to that of control-iPSC-CMs, by assessing their morphology, hypertrophy-related genes expression, calcium handling, diastolic function and myofilament calcium sensitivity at days 15 and 40 respectively. Finally, to better understand increased myofilament Ca2+ sensitivity as a central mechanism of central pathogenicity in HCM, inhibition of calcium sensitivity with mavacamten can improveed cardiomyocyte hypertrophy. Mutant-iPSC-CMs exhibited enlarged areas, increased sarcomere disarray, enhanced expression of hypertrophy-related genes proteins, abnormal calcium handling, diastolic dysfunction and increased myofilament calcium sensitivity at day 40, but only significant increase in calcium sensitivity and mild diastolic dysfunction at day 15. Increased calcium sensitivity by levosimendan aggravates cardiomyocyte hypertrophy phenotypes such as expression of hypertrophy-related genes, abnormal calcium handling and diastolic dysfunction, while inhibition of calcium sensitivity significantly improves cardiomyocyte hypertrophy phenotypes in mutant-iPSC-CMs, suggesting increased myofilament calcium sensitivity is the primary mechanisms for MYH7 mutations pathogenesis. Our studies have uncovered a pathogenic mechanism of HCM caused by MYH7 gene mutations through which enhanced myofilament calcium sensitivity aggravates abnormal calcium handling and diastolic dysfunction. Correction of the myofilament calcium sensitivity was found to be an effective method for treating the development of HCM phenotype in vitro.

PLEKHM2 deficiency induces impaired mitochondrial clearance and elevated ROS levels in human iPSC-derived cardiomyocytes.

Pleckstrin homology domain-containing family M member 2 (PLEKHM2) is an essential adaptor for lysosomal trafficking and its homozygous truncation have been reported to cause early onset dilated cardiomyopathy (DCM). However, the molecular mechanism of PLEKHM2 deficiency in DCM pathogenesis and progression is poorly understood. Here, we generated an in vitro model of PLEKHM2 knockout (KO) induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to elucidate the potential pathogenic mechanism of PLEKHM2-deficient cardiomyopathy. PLEKHM2-KO hiPSC-CMs developed disease phenotypes with reduced contractility and impaired calcium handling. Subsequent RNA sequencing (RNA-seq) analysis revealed altered expression of genes involved in mitochondrial function, autophagy and apoptosis in PLEKHM2-KO hiPSC-CMs. Further molecular experiments confirmed PLEKHM2 deficiency impaired autophagy and resulted in accumulation of damaged mitochondria, which triggered increased reactive oxygen species (ROS) levels and decreased mitochondrial membrane potential (Δψm). Importantly, the elevated ROS levels caused oxidative stress-induced damage to nearby healthy mitochondria, resulting in extensive Δψm destabilization, and ultimately leading to impaired mitochondrial function and myocardial contractility. Moreover, ROS inhibition attenuated oxidative stress-induced mitochondrial damage, thereby partially rescued PLEKHM2 deficiency-induced disease phenotypes. Remarkably, PLEKHM2-WT overexpression restored autophagic flux and rescued mitochondrial function and myocardial contractility in PLEKHM2-KO hiPSC-CMs. Taken together, these results suggested that impaired mitochondrial clearance and increased ROS levels play important roles in PLEKHM2-deficient cardiomyopathy, and PLEKHM2-WT overexpression can improve mitochondrial function and rescue PLEKHM2-deficient cardiomyopathy.

Generation of a TRPM8 knockout hESC line (WAe009-A-A) derived from H9 using CRISPR/Cas9.

The transient receptor potential cation channel subfamily M member 8 (TRPM8) is a kind of non-selective cation channel which controls Ca2+ homeostasis. Mutations in TRPM8 were related to dry eye diseases (DED). Here we constructed a TRPM8 knockout cell line WAe009-A-A from the original embryonic stem cell line H9 using CRISPR/Cas9 technology, which maybe helpful for exploring the pathogenesis of DED. WAe009-A-A cells possess stem cell morphology and pluripotency as well as normal karyotype, and have the ability of differentiating into three germ layers in vitro.

Severe fever with thrombocytopenia syndrome virus replicates in platelets and enhances platelet activation.

BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS) virus (SFTSV) infection causes an emerging hemorrhagic fever in East Asia with a high mortality rate. Thrombocytopenia is a consistent feature of SFTS illness, but the mechanism remains elusive. OBJECTIVES: We aimed to better understand the role of platelets in the pathophysiology of SFTSV infection, including the development of thrombocytopenia. METHODS: Using platelets from healthy volunteers and patients with SFTS, we evaluated the functional changes in platelets against SFTSV infection. We investigated the direct effect of glycoprotein VI on platelet-SFTSV interaction by quantitative real-time PCR, molecular docking, surface plasmon resonance spectrometry, flow cytometry, western blot, and platelet functional studies in vitro. Interactions of SFTSV and platelet-SFTSV complexes with macrophages were also determined by scanning electron microscope, quantitative real-time PCR, and flow cytometry. RESULTS: This study is the first to demonstrate that platelets are capable of harboring and producing SFTSV particles. Structural and functional studies found that SFTSVs bind platelet glycoprotein VI to potentiate platelet activation, including platelet aggregation, adenosine triphosphate release, spreading, clot retraction, coagulation, phosphatidylserine exposure, thrombus formation, and adherence. In vitro mechanistic studies highlighted that the interaction of platelets with human THP-1 cells promoted SFTSV clearance and suppressed cytokine production in macrophages. However, unwanted SFTSV replication in macrophages reciprocally aggravated SFTSV persistence in the circulation, which may contribute to thrombocytopenia and other complications during SFTSV infection. CONCLUSION: These findings together highlighted the pathophysiological role of platelets in initial intrinsic defense against SFTSV infections, as well as intertwined processes with host immunity, which can also lead to thrombocytopenia and poor prognosis.

Association Between Serum Aldehydes and Hypertension in Adults: A Cross-Sectional Analysis of the National Health and Nutrition Examination Survey.

Background: Exposure to ambient pollutants and chemicals were found to be associated with increased risk of hypertension. However, the relationship between the increased aldehyde exposure and hypertension are still unclear. This study aimed to investigate the potential associations of serum aldehydes levels with prevalent hypertension. Methods: A total of 1,733 U.S. adults with data on hypertension outcome and serum aldehydes measurement from the National Health and Nutrition Examination Survey 2013-2014 were included. The serum levels of aldehydes were measured via an automated analytical method using solid phase microextraction gas chromatography and high-resolution mass spectrometry. Multivariate logistic regression models were adopted to assess the associations between six selected aldehydes exposure (benzaldehyde, butyraldehyde, heptanaldehyde, hexanaldehyde, isopentanaldehyde, and propanaldehyde) and prevalence of hypertension. Results: The mean age was 48.0 ± 16.7 years and an approximately equivalent of sex distribution was observed (female 49.9%). There seems to be a numerically higher level of hexanaldehyde in participants with hypertension when compared to participants without hypertension (2.6 ± 3.9 ng/mL vs. 2.3 ± 1.1 ng/mL). After adjusting for potential confounders, the odds ratio (OR) for hypertension was 2.15 [95% confidence interval (CI): 1.33-3.51] in participants from the highest quartile of serum hexanaldehyde concentration in comparison to those from the lowest quartile. Subgroup analyses and sensitivity analyses showed generally similar results. Conclusion: In summary, current evidence suggested that increased serum hexanaldehyde level was positively associated with prevalent hypertension in U.S. adults.

Generation of an iPSC line (ZZUNEUi021-A) from a hypertrophic cardiomyopathy patient with TNNT2 mutation.

A 25-years-old hypertrophic cardiomyopathy male patient donated his peripheral blood mononuclear cells (PBMCs) with heterozygote mutation in theTNNT2 gene. We generated induced pluripotent stem cell (iPSC) with normal karyotypic and expressing NANOG, Lin28, GDF3 and DNMT3. The iPSC line has demonstrated pluripotency by differentiating into three germ layers in vitro. The ZZUNEUi021-A would serve as an in vitro model for loss of TNNT2 function.

Generation of an iPSC line (ZZUNEUi016-A) derived from a hypertrophic cardiomyopathy patient with the heterozygote mutation in MYH7 gene.

HCM is one of the most common inheritable cardiac disease. In our study, we established a human-induced pluripotent stem cells (hiPSCs) line (ZZUNEUi016-A) from a hypertrophic cardiomyopathy patient with the pathogenic heterozygote mutation in MYH7 gene. ZZUNEUi016-A expressed pluripotency markers with normal karyotype and showed the ability to differentiate into all three germ layers in vitro.

A heterozygous MYH7 (c. 2156G > A) mutant human induced pluripotent stem cell line (ZZUNEUi020-A) generated from a patient with hypertrophic cardiomyopathy.

Hypertrophic cardiomyopathy (HCM) is a heterogeneous myocardial disease often caused by sarcomeric gene mutations. MYH7 is one of the most common genes associated with HCM. In this study, we generated a human induced pluripotent stem cell (iPSC) line ZZUNEUi020-A from peripheral blood mononuclear cells of a female HCM patient with the p. R719Q (c. 2156G > A) mutation in MYH7. This cell line expressed pluripotency markers, showed normal female karyotype and could differentiate into all three germ layers in vitro.

An induced pluripotent stem cells line (ZZUNEUi022-A) derived from urine cells of healthy male human.

Urine cells (or renal tubular cells) can be isolated from human urine samples efficiently. This noninvasive and cost-effective method to collect biological sample provide us favorable access to donor cells from human. In the present study, we generate ZZUNEUi022-A, a urine cells-derived induced pluripotent stem cell (iPSC) line, from a 29-year-old healthy male via Sendai virus delivery system. ZZUNEUi022-A showed stable karyotype, and could differentiate into three germ layers (ectoderm, mesoderm, and endoderm) readily in an embryoid body formation model.

Generation of a hiPSC line ZZUNEUi017-A from a patient with dilated cardiomyopathy caused by mutation in TTN.

Dilated cardiomyopathy (DCM) is the commonest type of cardiomyopathy. In this study, peripheral blood mononuclear cells (PBMCs) were isolated from a DCM patient with the p. Glu12513fs(c.37537delG) mutation in TTN and were reprogrammed to human induced pluripotent stem cells (iPSCs). The ZZUNEUi017-A iPSC line expressed pluripotency markers, exhibiting a normal male karyotype (46, XY) and demonstrating differentiation potential into three germ layers in vitro.

Generation of a human iPSC line ZZUNEUi015-A from a patient with hypertrophic cardiomyopathy caused by mutation in ALPK3.

Hypertrophic cardiomyopathy is the commonest monogenic cardiomyopathy in humans and was reported to be associated with ALPK3 gene mutation. We report the generation and characterization of the human induced pluripotent stem cell (iPSC) line ZZUNEUi015-A, which was derived from a patient with a heterozygous mutation in ALPK3 gene (c.1013 T > C) and diagnosed with hypertrophic cardiomyopathy. The ZZUNEUi015-A line maintains the morphology of stem cells, has pluripotency and normal karyotype, and differentiated into three germ layers in vitro.

Induced pluripotent stem cell (iPSC) line (ZZUNEUi009-A) from a healthy female individual.

Induced pluripotent stem cells (iPSCs) can be used to generate different types of somatic cells in vitro and are a useful tool for investigating drug and disease mechanisms. Here, we generated human induced pluripotent stem cell (iPSC) line ZZUNEUi009-A from an apparently healthy 28-year-old female by reprogramming peripheral blood mononuclear cells with non-integrating vector. The generated iPSCs was pluripotent, maintained a stable karyotype, and could generate the three layers (ectoderm, mesoderm, and endoderm) in vitro.

Induced pluripotent stem cell line (ZZUNEUi011-A) derived from peripheral blood mononuclear cells (PBMCs) of a healthy 27-year-old female individual.

In this study, we report a human induced pluripotent stem cell (iPSC) line from a healthy 27-year-old female individual using non-integrative Sendai viral reprogramming technology. The cell line expresses stemness markers, exhibits a normal female karyotype, and can differentiate into three germ layers in vivo. This iPSC line from a healthy individual provides a control group for studying disease mechanisms, drug screening, and toxicity testing.

Generation of a hiPSC line ZZUNEUi012-A from a healthy female individual.

Induced pluripotent stem cells (iPSCs) have differentiation potential into different somatic cell types in vitro and are a useful tool to investigate pathomechanistic and cellular processes. In this study, we generated human induced pluripotent stem cells (iPSC) ZZUNEUi012-A from an apparently healthy female individual using an integration-free reprogramming method. The generated hiPSC line was pluripotent and had normal karyotype, showed robust expression of pluripotency markers and could differentiate into all three germ layers in vitro.

Generation of an IPSC line from a patient with hypertrophic cardiomyopathy carrying a mutation in MYH6 gene.

An induced pluripotent stem cell (iPSC) line was generated from peripheral blood mononuclear cells (PBMCs) of a 41-year-old male patient with hypertrophic cardiomyopathy who carries a G3755A heterozygote mutation in the MYH6 gene. The generated iPSC line expressed pluripotency markers, exhibited a normal karyotype, presented the specific mutation, and demonstrated differentiation potential into three germ layers in vitro.

Drug-coated balloon angioplasty: predicting outcomes based on different patterns of drug-eluting stent restenosis.

Although drug-coated balloon (DCB) angioplasty is an effective therapy for drug-eluting stent- in stent restenosis (DES-ISR) after coronary stenting, recurrent ISR after DCB angioplasty still occurs. Different patterns of DES-ISR responding to DCB are largely unknown. This study sought to assess outcomes of different patterns of DES-ISR treated with DCB. From December 2014 to December 2016, a total of 160 DES-ISR lesions treated with DCB were retrospectively evaluated. Restenosis patterns were classified into two groups according to Mehran classification: focal, defined as < 10 mm, 58 lesions (36.3%); non-focal, which were diffuse, proliferative, or obstructive, 102 lesions (63.7%). The primary endpoint was binary restenosis rate at 9-month angiographic follow-up. Secondary endpoint was major adverse cardiac events (MACE) at 24-month follow-up. Baseline characteristics were comparable between the two groups. Angiographic follow-up rate was 93.7% (93.1% in the focal group and 94.1% in the non-focal group). The focal group had a lower recurrent restenosis rate compared to the non-focal group (3.7% vs. 33.3%, respectively; P = 0.003) at an average angiographic follow-up of 10 (10.4 ± 6.2) months. There was no difference in MACE between the two groups (6.9% vs. 11.8%, respectively; P = 0.70) at (22.7 ± 9.1) months clinical follow-up. On multivariate logistic regression analysis, focal pattern (OR 13.033; 95% CI 2.441-69.573, P = 0.003) and post-procedure DS% (OR 1.142; 95% CI 1.070-1.218, P = 0.000) were predictive factors of binary restenosis after DCB angioplasty. On multivariate analysis, focal pattern of ISR was a predictive factor of MACE (OR 0.260; 95% CI 0.071-0.959, P = 0.043), and diabetes mellitus (DM) was an independent predictor of MACE after DCB angioplasty (OR 5.045; 95% CI 1.179-21.590, P = 0.029). The present study suggests that DCB provides much better clinical, angiographic outcomes in patients with focal DES-ISR than non-focal DES-ISR.

Generation of a hiPSC line ZZUNEUi007-A from a patient with hypertrophic cardiomyopathy caused by mutation in MYH7.

Hypertrophic cardiomyopathy (HCM) is the most common monogenic cardiovascular disorder. In this study, we generated human induced pluripotent stem cells (iPSC) ZZUNEUi007-A from dermal fibroblasts of an HCM patient with the p. R663H (c. 1988 G > A) mutation in MYH7. The generated hiPSC line had normal karyotype, showed robust expression of pluripotency markers and could differentiate into all three germ layers in vivo.

Effect of albumin-globulin score and albumin to globulin ratio on survival in patients with heart failure: a retrospective cohort study in China.

OBJECTIVE: To investigate the combined effect of albumin (ALB) and globulin (GLB) on the overall survival (OS) of patients with heart failure (HF). DESIGN: Retrospective cohort study. SETTING: A hospital. PARTICIPANTS: 404 patients first diagnosed with HF. MEASUREMENTS: Serum ALB and GLB were measured within 3 days after admission. The albumin to globulin ratio (AGR) was calculated as the ALB divided by the GLB. The receiver operating characteristic curve was used to calculate the cut-off points for ALB, GLB and AGR. Patients with low ALB levels (≤35.3 g/L) and high GLB levels (>27.0 g/L) were assigned an albumin-globulin score (AGS) of 2, those with only one of the two abnormalities were assigned an AGS of 1 and those with neither of the two abnormalities were assigned an AGS of 0. RESULTS: The mean age of the 404 patients was 62.69±15.62, and 54.5% were male. 14 patients were lost to follow-up. 120 patients died from HF and 211 patients were readmitted to the hospital for worsening HF. Multivariate Cox regression analysis showed that higher AGR was significantly associated with favourable OS (HR, 0.61, 95% CI 0.38 to 0.98, p=0.040) but not AGS. CONCLUSION: Serum levels of ALB and GLB are objective and easily measurable biomarkers which can be used in combination to predict the survival of patients with HF.