bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cardiac sympathetic afferent denervation reduces matrix metalloproteinase expression and improves cardiac remodeling in rats post myocardial infarction
Authors: Zucker I , Wang H , Mishra P K , Nandi S 
Published: October 2022

Abstract

In a previous study from our laboratory, we demonstrated that chronic and selective cardiac sympathetic afferent (spinal) denervation at the time of myocardial infarction (MI) using epicardial application of resiniferatoxin (RTX), a neuronal toxin capable of inducing rapid degeneration of transient receptor potential vanilloid 1 (TRPV1)-expressing afferent neurons and fibers, markedly reduced the cardiac remodeling process 9–11 weeks post-MI in rats. This included reduced cardiac hypertrophy, fibrosis and apoptosis. Here, we further investigated the effect of epicardial application of RTX at the time of MI on cardiac extracellular matrix (ECM) and remodeling post-MI. Echocardiographic and morphologic data demonstrated that, compared to MI+vehicle, MI+RTX exhibited a significantly slower LV chamber dilation (6-week echocardiographic data: left ventricular end-diastolic diameter, 10.7±0.2 vs. 9.6±0.3 mm; left ventricular end-systolic diameter: 8.8±0.2 vs. 7.8±0.3 mm; MI+vehicle vs. MI+RTX, n=18, p&lt;0.05; mean±SE). Scanning electron microscopy showed that RTX reduced collagen deposition in the peri-infarct area in post-MI rats. Western blot and zymography were used to further evaluate the effect of RTX application on matrix metalloproteinase (MMP) expression and activity, which is responsible for degrading ECM and contributing to cardiac dilation post MI. Our data suggest time-dependent increases in MMP expression in infarcted hearts post MI. RTX application largely prevented the increase in MMP9 but not MMP2, 4 weeks post-MI. RTX prevented both increased MMP2 and MMP9 activities in the peri-infarct myocardium at 8–10 weeks post-MI. We further investigated the effect of RTX application on MI-induced cardiac inflammation, which has been reported as an upstream mechanism triggering MMP activation post-MI. The data show that RTX largely abolished MI-induced plasma extravasation and reduced macrophage infiltration and cardiac cytokine content in the peri-infarct or remote myocardium of post-MI rats. These data suggest that cardiac sympathetic afferent denervation at the time of MI exerts a local anti-inflammatory effect and reduces ECM remodeling by preventing excessive MMP activation in post-MI rats.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): National Institutes of Health Grant HL126796
Modeling cardiac fibroblast heterogeneity in fibrotic heart using human pluripotent stem cell derived epicardial cells
Authors: Funakoshi S , Fernandes I F , Keller G K 
Published: October 2022

Abstract

Introduction
Our understanding of human cardiac pathophysiology is limited due to the scarcity of human heart samples. Model organisms do not faithfully recapitulate human physiology and pathophysiology to the extent that is needed. Therefore, human pluripotent stem cell (hPSC)-derived cardiac cells represent promising sources to overcome these limitations to studying heart diseases.

Purpose
To generate cardiac organoids mimicking the interaction between cardiomyocytes and non-cardiomyocytes in the developing heart and the postnatal pathological conditions.

Method
It has been well known that epicardium is a main source of non-cardiac cells in the developing heart. We therefore generated cardiac organoids by mixing hPSC-derived cardiomyocytes and hPSC-derived epicardium and test if they could mimic the developing heart and be applicable for modeling heart failure.

Results
We established a cardiac organoid system generated from human pluripotent stem cells that models the developing heart's early interactions, specifically the epicardial and non-myocyte development, as well as ventricular cardiomyocyte proliferation. Inside the cardiac organoids, epicardium spontaneously differentiated into CD90-positive cardiac fibroblasts expressing fibroblast markers, FN, COL1 and COL3, and CD90-negative smooth muscle cells (SMC) expressing SMC markers, ACTA2 and MYH11. Additionally, we observed the significant increase in cardiomyocyte proliferation in the cardiac organoids compared to in cardiomyocyte only aggregation (p&lt;0.001). We then mature our cardiac organoids using our unique maturation method, the combination of PPARα agonist, dexamethasone, T3 hormone, and palmitate, generating metabolically mature cardiac organoids. These mature organoids recapitulated the phenotypic changes observed in the failing human heart when exposed to pathological stimuli, including fibrosis, metabolic changes, and increase in heart failure markers. We used single-cell transcriptomics to dissect the cellular heterogeneity of our organoids in a model of heart failure with comparisons to primary human data and revealed our organoids recapitulated in vivo human failing heart. This analysis allowed to confirm that our organoids contain a unique subpopulation of cardiac fibroblasts possessing reparative features, suggesting the recapitulation of in vivo heterogeneity in our model.

Conclusions
We successfully generated cardiac organoids mimicking the interaction between cardiomyocytes and non-cardiac cells. Our system enables the recapitulation of in vivo heterogeneity, providing a platform for the accurate understanding of heart development and disease in a dish.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Canadian Institutes of Health Research (CIHR)
Human epididymis protein 4 indicates progression of cardiac fibrosis in Heart Failure with preserved Ejection Fraction
Authors: Nakashima N , Hanatani S H , Yamamoto M Y , Ogata F O , Araki S A , Arima Y A , Matsushita K M , Tsujita K T 
Published: October 2022

Abstract

Background
Cardiac fibrosis is one of the main causes of diastolic dysfunction in heart failure with preserved ejection fraction (HFpEF). Human epididymis protein 4 (HE4) is a novel pro-fibrotic protein expressed especially in activated fibroblast, so-call myofibroblast [1]. Although our previous study showed that HE4 functioned as a secretory factor promoting cardiac fibrosis in dilated cardiomyopathy [2], little is known about the role of HE4 in HFpEF.

Purpose
The aim of this study is to evaluate the pathophysiological role of HE4 in HFpEF and the association between serum HE4 levels and clinical course of HFpEF.

Methods
In basic research, we administered high-fat diet and NOS inhibitor (L-NAME) to mice for 5 weeks to induce HFpEF. We analyzed echocardiographic findings and mRNA expressions of fibrosis-related genes including HE4 by real time PCR. In clinical research, we measured serum HE4 levels of 79 patients with HFpEF and divided these patients into low and high HE4 group using median values of HE4 (102.0 pmol/L). Then, we evaluated their mortality and cardiovascular events retrospectively.

Results
In vivo, the administration of high fat diet and L-NAME induced significant increase in left ventricular mass on echocardiography compared with control group (p=0.006). The mRNA expressions of aSMA and collagen3a1 in heart tissue was upregulated in HFpEF model mice (p=0.008 and p=0.003, respectively). Furthermore, the mRNA expression of HE4 was also upregulated in HFpEF model mice (n=6) than in control mice (n=5), although these differences did not reach statistical significance (p=0.238). In vitro, recombinant HE4 upregulated mRNA expression of pro-fibrotic genes, such as periostin (p=0.001), collagen3a1 (p=0.037), aSMA (p=0.095), and PAI-1 (p=0.17), indicating HE4 facilitate the activation of fibroblast and cardiac fibrosis although some did not reach statistical significance.
In clinical research, Kaplan-Meier curve revealed that the rate of all cause death and rehospitalization for worsening heart failure were significantly higher in the high than the low HE4 group, suggesting HE4 can predicts prognosis in patients with HFpEF.

Conclusion
HE4 induces the activation of cardiac fibroblasts and is upregulated in the heart of HFpEF model mice. Furthermore, it is a useful biomarker for predicting cardiovascular events in patients with HFpEF.

Funding Acknowledgement
Type of funding sources: None.
NLRP3 mediates atheromatous plaque macrophage proliferation
Authors: Haerdtner C , Remmersmann F , Von Ehr A , Dederichs T S , Vico T , Krebs K , Wolf D , Sager H , Bode C , Westermann D , Hilgendorf I 
Published: October 2022

Abstract

Background
Macrophage accumulation in atherosclerotic plaques drives disease progression largely dependent on in situ proliferation. We previously reported that systemic cholesterol lowering or reduced modified lipoprotein uptake supress atheromatous plaque macrophage proliferation. In this work we investigate the intracellular mediators of macrophage proliferation.

Methods and results
Macrophages deficient in scavenger receptors CD36 or Msr1 and impaired in cholesterol-rich lipoprotein uptake, proliferated less compared to control macrophages in the same plaque exposed to the same lipid levels in a LDLR-deficient bone marrow irradiation mixed chimera model. Proliferation of plaque macrophages deficient in the intracellular cholesterol sensor LXR was not impaired, however. As modified LDL uptake can activate the NLRP3 inflammasome, we generated mixed bone marrow knockout chimeras for components of the inflammasome. NLRP3 but not Caspase-1 or interleukin-1 receptor deficient macrophages proliferated 35% less compared to NLRP3-expressing macrophages in the same plaque in vivo. These results were confirmed in NLRP3-deficient oxLDL stimulated macrophages in vitro. In line, NLRP3 inhibition of human carotid artery plaque cultures suppressed human plaque macrophage proliferation and reduced inflammasome-dependent IL-1b secretion. However, IL-1b supplementation did not restore local macrophage proliferation in accord with our findings in IL-1 receptor deficient murine plaque macrophages.

Conclusion
We identified a novel role for NLRP3, independent of the canonical Caspase-1–IL-1b inflammasome pathway, in mediating macrophage proliferation in atherosclerotic plaques in mice and men representing a druggable target.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): DFG
Injectable carbon nanotube-functionalized hydrogel as a tool for cardiac tissue engineering
Authors: Pena B , Knight W , Cavasin M , Ferrari I , Abdel-Hafiz M , Vagnozzi R , Bosi S , Park D , Shandas R , Song K , McKinsey T , Sbaizero O , Taylor M , Prato M , Mestroni L 
Published: October 2022

Abstract

Background/Introduction
Heart failure (HF) is an expensive major public health problem in the United States and around the world (1). The current treatments for HF are aimed at reducing symptoms, slowing disease progression, and reducing mortality and not aimed at repairing heart muscle or restoring function. Furthermore, even with these treatments, approximately half of patients with HF will die within 5 years of diagnosis (2). Cardiac transplantation remains the only definitive treatment for those affected with end-stage HF, but availability of donor hearts remains a major limitation (3).

Purpose
The ability of the adult heart to regenerate cardiomyocytes (CMs) lost after injury is limited, generating interest in developing tissue engineering therapies to avoid progression towards HF. Rigid carbon nanotubes (CNTs) scaffolds have been used to improve CMs viability, proliferation, and maturation (4), but require undesirable invasive surgeries for implantation. To overcome this limitation, we engineered an injectable reverse thermal gel (RTG) functionalized with CNTs (RTG-CNT) that transitions from a liquid-solution to a gel-based matrix shortly after reaching body temperature allowing for a liquid-based delivery rapidly followed by a stable-gel localization (5).

Methods and results
Here we show experimental evidences the RTG-CNT hydrogel, used as a three-dimensional (3D) niche to culture human induced pluripotent stem cells (hiPSC)-CMs, promotes hiPSC-CMs alignment and elongation with increased Cx43 localization and improved contraction function when compared with traditional two-dimensional (2D) fibronectin controls and plain 3D RTG system without CNTs. Moreover, the short-term (4-week) biocompatibility of the RTG-CNT hydrogel was also assessed in a mouse model (intracardial injection). The results confirmed that the RTG-CNT hydrogel is well tolerated by the cardiac tissue.

Conclusion
Our results indicated that the injectable RTG-CNT hydrogel has the potential to be used as a minimally invasive tool for cardiac tissue engineering efforts.

Funding Acknowledgement
Type of funding sources: Other. Main funding source(s): NATIONAL HEART, LUNG, AND BLOOD (NHLBI) INSTITUTE
Correlation between coronary artery disease and coronary microvascular dysfunction symptoms and cardiac SPECT results in Latvian women patients
Authors: Vitola B , Mintale I , Trusinskis K , Kalnina M , Kedrova J , Erglis A 
Published: October 2022

Abstract

Introduction
The clinical evaluation of symptomatic women is challenging due to their varying clinical presentation. Women patients with intermediate or high pre-test probability for coronary artery disease – even asymptomatic or with non-informative exercise test should be referred for further focused diagnostics. Myocardial perfusion imaging with single photon emission computed tomography (SPECT) is one of the most performed non-invasive cardiac imaging tests to diagnose CAD or coronary microvascular dysfunction (CMVD).

Purpose
To evaluate the correlation between various CAD symptoms and cardiac SPECT results in women patients undergoing examination due to diagnosis of CAD or CMVD.

Methods
During the period from 2018 till 2021 we performed cardiac SPECT for women patients with intermediate or high pre-test probability or non-informative exercise test. Patients were divided into 2 groups: normal myocardial perfusion or true myocardial perfusion defect, defined as area of ischemia ≥10% of the left ventricle myocardium mass. According to the clinical symptoms patients were categorized in “asymptomatic”, “atypical symptom” and “patients with chest pain” group. The presence of CMVD and CAD in each group was determined by invasive coronary angiography or computed tomography angiography.

Results
In 4 years, SPECT was performed in 726 women patients. The most common presented symptoms were: chest pain (n=487, 67.1%), dyspnea (n=323, 44.5%), tachycardia and palpitations (n=117, 16.1%), fatigue (n=129, 17.8%), headache (n=28, 3.86%) and excessive sweating (n=15, 2.1%). 104 (14.3%) patients were asymptomatic, 487 (67.1%) with chest pain and 135 (18.6%) with atypical symptoms. True myocardial perfusion defect was detected in 125 patients (17.2%). The obstructive CAD was found in 32 cases (25.6%; 4.4% of all SPECTs). The CMVD was diagnosed in 68 women (54.4%; 9.4% of all SPECTs). In asymptomatic patient group obstructive CAD was diagnosed in 5 (4.8%) cases, in chest pain group – 11 (2.3%) cases and in atypical symptom group – 16 (11.9%) patients (p&lt;0.001). None of asymptomatic women had CMVD. The CMVD was detected in 53 (42.4%; 7.3% of all) cases in chest pain group and in 14 (11.3%; 1.9% of all) patients in atypical symptom group (p=0.006).

Conclusion
Women patients with typical chest pain were more likely to have CMVD, but with atypical symptoms – CAD. Since women are in a very high cardiovascular risk group in Latvia, the cardiac SPECT is highly recommended for detection of CAD and CMVD, especially in patients with atypical symptoms.

Funding Acknowledgement
Type of funding sources: None.
Live imaging of nitric oxide release in vascular endothelial cells in response to mechanical stimuli on an organ chip
Authors: Takahashi K , Liu Y , Wang M , Liang Y , Naruse K 
Published: October 2022

Abstract

Background
Nitric oxide (NO), released from vascular endothelial cells in response to mechanical stimuli, regulates cardiac contractility and are also involved in the prevention of the development of cardiac hypertrophy.

Purpose
To establish an experimental system for live observation of NO release in response to mechanical stimuli on an organ chip.

Methods
Organ chips, which we used for the development of a heart-on-a-chip in the previous study [1], were used.
We seeded 300,000 human umbilical vein endothelial cells on a stretchable elastic membrane coated with Matrigel of a chip channel. Shear stress was applied to the cells by increasing flow rate of a peristaltic pump connected to the chip channel (Figure 1A). Pressure stimulus was applied by hydrostatic pressure. Stretch stimulus was applied by suction to the side ports of a chip using an electric syringe pump (Figure 1B). Cells were stained with 10 μM 4,5-diaminofluorescein diacetate for fluorescent live NO imaging.

Results
Monolayers of the endothelial cells formed intercellular junctions confirmed by CD31 staining (Figure 1C, yellow). Apparent permeability, which was measured by Texas red dye (MW 3000), was maintained at a low level of ∼3x10–6 cm/s until day 30, suggested the formation of robust intercellular junction.
When the endothelial cells were subjected to a pressure stimulus of 60 mmHg for 60 s, NO release was observed that lasted for &gt;2 minutes (Figure 2A). A peak value of 1.46±1.08 (mean ± standard deviation) times the baseline was observed 271 s after the beginning of the pressure stimulus (n=251 cells). When the cells were subjected to a 1% stretch for 60 s, a peak value of 1.29±0.33 times the baseline was observed 105 s after the beginning of the stretch stimulus (Figure 2B). A shear stress of 0.01 dyn/cm2 hardly increased NO release (1.20±0.27 times the baseline, Figure 2C).

Conclusion
The system for live NO imaging in vascular endothelial cells in response to mechanical stimuli was established using organ-on-a-chip. The heart-on-a-chip with endothelial cells will be useful in elucidating the effects of mechanical stimulus such as hypertension on the contractile function and the remodeling of the heart.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Japan Society for the Promotion of Science
Comparative prognostic utility of visual grade and heart-to-contralateral lung ratio in patients undergoing 99mTc-pyrophosphate radiotracer cardiac scintigraphy
Authors: Saad J M , Ahmed A I , Alahdab F , Han Y , Alfawara M , Kassi M , Al-Mallah M H 
Published: October 2022

Abstract

Introduction
Transthyretin (ATTR) cardiac amyloidosis (CA) is an underdiagnosed form of restrictive cardiomyopathy leading to a rapid progression into heart failure. 99mTc-labeled radiotracer cardiac scintigraphy is crucial for the evaluation of CA. Semiquantitative (Perugini visual grade) and quantitative (heart-to-contralateral lung ratio, H/CL) parameters are used to assessing CA via cardiac scintigraphy.

Purpose
We aimed to assess the comparative prognostic utility of the Perugini visual grade and heart to contralateral lung ratio.

Methods
The study population was identified based on an institutional registry of consecutive patients undergoing 99mTc-PYP radiotracer cardiac scintigraphy for suspected CA between January 2020-October 2021. The H/CL is calculated by the fraction of heart region of interest (ROI) mean uptake counts to the contralateral chest ROI. The visual scoring system compares uptake between bone (rib) and heart where 0 = absent cardiac uptake, 1 = uptake less than bone, 2 = uptake equal to bone, and 3 = is uptake greater than bone. Ejection fraction (EF) was obtained from echocardiographic studies done around the time of cardiac scintigraphy. MACE was defined as a composite of inclusive of all-cause death, myocardial infarction, admission for heart failure and late revascularization – percutaneous coronary intervention (PCI) or coronary artery bypass graft (CABG) &gt;90 days after imaging.

Results
A total of 330 patients (mean age 70.9±12.2 years, 62.2% male) were included in the analysis. Risk factors were prevalent (59.7% hypertension, Heart Failure 68.3%, 26.5% dyslipidemia, 26.2% diabetes) (Table 1). In total, 32.6% of the cohort had studies suggestive of ATTR CA. Median time between echocardiography and cardiac scintigraphy was 7 days (IQR 5–42 days). A higher frequency of MACE was reported in patients with a 2+ visual grade (30% vs 42%, p=0.03). A H/CL &gt;1.5 was associated with borderline significance (43% vs 35% p=0.07).

Conclusion
Our analysis showed that the Perugini visual grading of transthyretin cardiac amyloidosis conferred the best prognostic utility contrary to heart to contralateral lung ratio in 99mTc-PYP radiotracer cardiac scintigraphy.

Funding Acknowledgement
Type of funding sources: None.
Effect of coagulation factors FXIa, FXIIa, F alpha-XIIa, F beta-XIIa and FXIIIa on platelet function
Authors: Wildeis L , Ziese J , Helten C , Benkhoff M , Mourikis P , Zako S , Petzold T , Levkau B , Zeus T , Kelm M , Polzin A 
Published: October 2022

Abstract

Background
FXa has recently been shown to active platelets via protease activated receptor. Direct inhibition of FXa leads to reduced platelet reactivity and arterial thrombosis. It is not known, if other coagulation factors besides FXa and FIIa directly activate platelets. In this study, we hypothesized that FXIa, FXIIa, Fα-XIIa, Fβ-XIIa and FXIIIa directly active platelets.

Purpose
Direct inhibitors of above mentioned factors are currently in clinical trials. Hence, investigation of coagulation factors' non-canonical effects beyond activation of the coagulation cascade are of great importance.

Methods
Platelet reactivity was measured using light transmission aggregometry (LTA) and flow cytometry. Ex-vivo stimulation of washed platelets was conducted using adenosine diphosphate (ADP) and thrombin receptor activator peptide (TRAP).

Results
FXIa, FXIIa, Fα-XIIa, Fβ-XIIa and FXIIIa have no direct effect on platelet function in terms of aggregation (Maximum of aggregation: FXIa: 2.799±1,823%, FXIIa: 1,174±1,080%, Fα-XIIa: 1,663±1,680%, Fβ-XIIa: 1,060±1,356%, Fα-XIIa + Fβ-XIIa 1,790±2,327% and FXIIIa: 1,345±1,060%). Also, incubation of platelets does not cause expression of P-selectin (FXIa: 4,346±4,650%, FXIIa: 7,072±6,098%, Fα-XIIa: 9,124±6,075%, Fβ-XIIa: 4,556±4,693%, Fα-XIIa + Fβ-XIIa 8,070±4,342% and FXIIIa: 5,166±5,512%).

Conclusion
FXIa, FXIIa, Fα-XIIa, Fβ-XIIa and FXIIIa have no direct effect on platelet function in terms of aggregation and P-selectin expression. Thus, existing pharmacological inhibitors of these factors are to be classified as purely anticoagulant and do not exert non-canonical effects on platelet activation.

Funding Acknowledgement
Type of funding sources: None.