bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immune checkpoint inhibition with PD-1 inhibitor induces cardiac dysfunction without overt myocarditis in C57BL/6J mice
Authors: Gergely T , Kucsera D , Toth VE , Petrovich B , Agg B , Onodi ZS , Ruppert M , Radovits T , Merkely B , Ferdinandy P , Varga ZV 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – EU funding. Main funding source(s): European Union’s Horizon 2020 Research and Innovation Programme under grant agreement no. 739593
“Semmelweis 250+ Kiválósági PhD Ösztöndíj” (EFOP-3.6.3-VEKOP-16-2017-00009)
Gedeon Richter Talentum Foundation’s scholarship

Background
Immune checkpoint inhibitors have revolutionized the treatment of several form of malignancies (including metastatic melanoma) by enhancing the cytotoxic effects of T cells against cancer cells. Cancer cells evade immune surveillence by increasing the expression of T cell inhibitory molecules, also known as immune checkpoints, such as programmed cell death-1 (PD-1). Pharmacological inhibition of these molecules by immune checkpoint inhibitors (ICI) will enhance the antitumor activity of T cells. However, enhanced T cell activity may cause immune related adverse effects, including cardiotoxicity.

Aims
We aimed to investigate the effect of PD-1 inhibition on cardiac function and the underlying mechanisms in mice.

Methods
8-10 weeks old C57BL6/J mice were treated with isotype control or anti-PD-1 antibody for 2 or 4 weeks. Cardiac function and morphology was assessed by echocardiography and histology, while the transcriptomic changes were analyzed via RNA sequencing. Nitrosative stress in the heart was assessed by immunohistochemistry and qRT-PCR. Inflammatory gene expression alterations were determined by qRT-PCR in the heart and thymus.

Results
Small animal echocardiography revealed cardiac dysfunction even after 2 weeks of anti-PD-1 treatment, with distinct transcriptomic changes. Nitrosative stress was found to be elevated in the myocardium due to anti-PD-1 treatment, however, histological and qRT-PCR analysis did not reveal T cell infiltration into the myocardium and only mild inflammation was seen in the heart. In contrast, inflammatory gene expression was significantly enhanced in the thymus of anti-PD-1-treated animals, where interleukin-17 showed the most prominent increase.

Conclusions
These findings characterize cardiac dysfunction as a form of ICI-induced cardiotoxicity, which may be mediated by increased thymic inflammatory activation and cytokine production.
Investigation of transcriptional changes underlying calcification of aortic valve
Authors: Semenova D , Zabirnyk A , Lobov A , Vaage J , Malashicheva A 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Foundation. Main funding source(s): The Russian Science Foundation

Introduction
Aortic valve stenosis due to calcification of valve cusps is the most common valve disease in the world today. The main feature of this condition is a progressive mineralization of valve tissue. The mechanisms underlying this process is still unknown, but in recent years it has become clear that pathological mineralization of heart and blood vessels has some similarities with the physiological process of bone formation. It has been suggested that interstitial cells (VICs) are the main functional units in the valve that undergo calcification. However, the early initiating mechanisms that trigger osteogenic transformation of cells remain unclear.

Purpose
The aim of the present study was to elucidate the most responsive time point of osteogenic differentiation induction and to identify the main osteogenic markers that mediate pathological calcification in human aortic valve.

Methods
VICs were obtained from patients with aortic valve calcification and from healthy aortic valves. The effectiveness of cell cultures osteogenic differentiation was estimated by Alizarin Red staining. Investigation of gene expression changes upon osteogenic differentiation was performed by qPCR and RNA sequencing.

Results
We found that 48 hours after the induction of osteogenic differentiation is the most relevant time point to identify the early regulators of osteogenic transformation of the cells. That is the time when the most intensive response from osteogenic markers takes place – BGLAP, OPG, OGN, RUNX2 – in comparison to 24, 72 and 96 hours of differentiation in both patient’s and healthy cells. We found out that induction of osteogenic differentiation on early stages initiates transcriptional program that serve to induce the next molecular events which recruit phenotype-specific osteogenes. We revealed that 558 and 232 genes which were up and down regulated during differentiation were the same for healthy and patient’s cells. However, there was a number of genes which was specific for either patient’s or healthy cells.

Conclusions
We presume that a great amount of the main molecular participants of osteogenic differentiation is shared between different types of cells which are prone to differentiation. However, we perform the results about specificity and difference between the mechanisms of osteogenic differentiation of patient’s and healthy cells.
Sex-specific cell types and molecular pathways drive fibro-calcific aortic valve stenosis
Authors: Myasoedova V , Massaiu I , Moschetta D , Chiesa M , Valerio V , Bozzi M , Parisi V , Poggio P 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Foundation. Main funding source(s): Fondazione Gigi e Pupa Ferrari ONLUS

Background
Aortic stenosis (AS) is the most common valve disorder characterized by fibro-calcific remodeling of valve leaflets. Recent evidence indicated that there is a sex-related difference in AS development and progression. Fibrotic remodeling is peculiar of women's aortic valve, while men's aortic leaflets are more calcified than in women.

Purpose
To assess aortic valve fibrosis (AVF) in a severe AS cohort using non-invasive diagnostic tools and determine whether sex-specific pathological pathways and cell types are associated with severe AS.

Methods
We have included 28 men and 28 women matched for age with severe AS who underwent Doppler echocardiography and cardiac contrast-enhanced computed tomography (CT) before intervention. The calcium and fibrosis volumes were assessed and quantified using the ImageJ thresholding method, indexed calcium and fibrosis volume were calculated by dividing the volume by the aortic annular area. Differentially expressed genes and functional inferences between women and men's aortic valves were carried out on a publicly available microarray-based gene expression dataset (GSE102249). Cell types enrichment analysis in stenotic aortic valve tissues was used to reconstruct the sex-specific cellular composition of stenotic aortic valves.

Results
We confirmed that women had significantly lower aortic valve calcium content compared to men, while fibrotic tissue composition was significantly higher in women than men. We identified that the expression profile of human stenotic aortic valves is sex-dependent. Pro-fibrotic processes were prevalent in women, while pro-inflammatory ones, linked to the immune response system, were enhanced in men. Cell-type enrichment analysis showed that mesenchymal cells were over-represented in AS valves of women, whereas signatures for monocytes, macrophages, T and B cells were enriched men ones.

Conclusions
Our data provide the basis that the fibro-calcific process of the aortic valve is sex-specific, both at gene expression and cell type level. The quantification of aortic valve fibrosis by CT could make it possible to perform population-based studies and non-invasive assessment of novel therapies to reduce or halt sex-related calcific aortic valve stenosis (CAVS) progression, acting in an optimal window of opportunity early in the course of the disease.
Paracrine signal emanating from stressed cardiomyocytes aggravates inflammatory microenvironment in diabetic cardiomyopathy
Authors: Kaur N , Ruiz A , Raja R , Howell G , Mace K , Binder P , Smith C , Swanton E , Wang X , Chernoff J , Cartwright EJ , Liu W 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Foundation. Main funding source(s): British Heart Foundation

Background
Cardiovascular issues associated with diabetes, such as diabetic cardiomyopathy (DCM), can lead to heart failure. DCM is etiologically related to myocardial inflammation and can stem from a complex interplay of different cell types. Cardiomyocyte as an active mediator of the inflammatory response is an emerging concept with limited mechanistic understanding.

Purpose
We aimed to address the knowledge gap of cardiomyocyte endoplasmic reticulum (ER) dysfunction-mediated macrophage response and provide functional evidence for the therapeutic feasibility of managing inflammatory paracrine signals in response to diet-induced metabolic stress.

Methods
In vivo mouse model of high fat high sucrose diet-induced diabetes, cardiomyocyte-specific p21-activated kinase 2 (PAK2) knockout model, echocardiography, histology, 3D imaging, qPCR, co-culture of H9c2 culturing medium and bone-marrow derived macrophages, immunoblotting, macrophage isolation from myocardium, flow cytometry and AAV9-gene therapy.

Results
In a time-course study, diet-induced diabetic mice demonstrated an association between cardiac ER stress and sustained myocardial inflammation, with a maladaptive shift in myocardial ER stress response over time. Furthermore, as a cardiac ER dysfunction model, mice with cardiac-specific PAK2 deletion exhibited heightened myocardial inflammatory response in diabetes. Using human and mice diabetic heart samples, we show that ER stress-induced CCAAT/enhancer-binding protein homologous protein (CHOP) is a novel transcriptional regulator of high mobility group box-1 (HMGB1). Cardiac stress-induced active release of HMGB1 facilitated M1 macrophage polarization, and aggravated myocardial inflammatory signatures. Therapeutically, sequestering the extracellular HMGB1 using Glycyrrhizin conferred cardioprotection through its anti-inflammatory action. Also, as functional evidence, we showed that un-mitigated cardiac ER response due to PAK2 loss under diabetes may account as a barrier for leveraging the anti-inflammatory potential of Vildagliptin.

Conclusion
Collectively, we introduce an ER stress-mediated cardiomyocyte-macrophage link, altering the macrophage response in the myocardium, thereby providing insight into therapeutic prospects for diabetes-associated cardiac dysfunction.
Tenascin-C provokes cardiac fibrosis and endothelial impairment in Duchenne Muscular Dystrophy
Authors: Szabo P L , Inci M , Hilber K , Abraham D , Trojanek S , Costantino S , Paneni F , Podesser B K , Kiss A 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): FWF Austrian Science Fund
Cardiac fibrosis and dilated cardiomyopathy are major contributors to mortality in Duchenne muscular dystrophy (DMD) patients. There is a growing collection of evidence that Tenascin C (TN-C) plays a maladaptive role in cardiac remodelling and fibrosis.
Our aims were to 1) assess the vascular dysfunction and cardiac fibrosis and its link to TN-C in a mouse model of DMD and 2) explore the effect of knocking out TN-C in dystrophic mice.
Male wt, mdx and mdx TN-C KO age-matched mice were used. Cardiac fibrosis was assessed on tissue sections. Wire myography was used to test the vascular reactivity and endothelial cells (ECs) were isolated from mouse lung tissues to characterize the oxidative stress and inflammatory marker expression. To study the signalling pathways contributing to cardiac fibrosis, human cardiac fibroblasts (hCFs) were treated with TN-C or TGF-β and gene expression and epigenetic regulation of p65 were assessed.
Cardiac fibrosis was markedly increased in mdx mice which was accompanied with elevated TN-C level in cardiac tissue and plasma compared to wt animals. In addition, endothelial cells isolated from mdx mice also showed a marked upregulation of oxidative stress and inflammatory markers and in line with that vascular endothelial function was impaired in mdx mice. Interestingly, mdx- TN-C KO mice showed preserved vascular function as well as reduced cardiac fibrosis compared to age-matched mdx mice. hCFs treated with TN-C or TGF-β showed increased collagen and α-SMA expressions which could be reduced by TN-C siRNA. In addition, both TN-C and TGF-β promote p65/NF-κB promoter demethylation and subsequently stimulate pro-inflammatory and pro-fibrotic signalling, which could be reversed by applying p38 MAPK inhibitor in hCFs.
TN-C promotes oxidative stress and inflammation in ECs and fibroblasts, contributing to severe endothelial dysfunction and cardiac fibrosis. In addition, activation of NF-κB p65 signalling pathway may play a role in TN-C induced fibrosis. Thus, TN-C may be a critical mediator and potential target for therapy in DMD.
Dysregulation of the endomembrane homeostasis triggers vascular smooth muscle cell phenotype switching
Authors: Hense N , Mir B , Marx N , Goettsch C 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Deutsche Forschungsgemeinschaft

Background
Cardiovascular diseases are a leading cause of mortality globally, with cardiovascular calcification as a prominent predictor and contributor. Notably, microcalcifications might cause atherosclerotic plaque rupture, leading to myocardial infarction. Calcified plaque remodeling accompanies phenotypic modulation of vascular smooth muscle cells (SMCs). Previously, we showed that extracellular vesicles (EVs) released from calcifying SMCs are pivotal for microcalcification formation. EVs originate from the endolysosomal system. The lipid kinase FYVE-type zinc finger containing phosphoinositide kinase (PIKfyve) plays a role in the endolysosomal maturation. Here, we investigated how modulation of the endolysosomal system intersects with SMC phenotype regulation.

Methods and Results
In calcified arteries and SMCs, we observed increased PIKfyve expression. Inhibition of PIKfyve with the small molecule Apilimod inhibited osteogenic differentiation of SMCs in an in vitro calcification model. Apilimod reduced tissue non-specific alkaline phosphatase (TNAP) – an early marker of SMC calcification on mRNA (-79%, p&lt;0.01), protein (-96%, p&lt;0.01) and activity (-92%, p&lt;0.01) levels. Additionally, matrix mineralization and collagen network formation are inhibited. SMC-secreted EVs exhibited a lower mineral content and reduced aggregation potential assessed by osteosense-based flow cytometry and turbidity assays. Transcriptome analyses revealed that the anti-calcifying effect is accompanied by a decreased expression of contractile SMC genes. Validation by RT-PCR supported that SMC-specific markers (ACTA2, SM22) and the key SMC transcription factor MYOCD were downregulated, while the expression of macrophage-like SMC markers (LGALS3, CD68) was induced. On the other hand, Apilimod increased the expression of adipogenic transcription factors (CEBPA, PPARG) and genes of the cholesterol and fatty acid metabolism pathways (SREBF2, HMGCR, FABP3, CD36) and induced the nuclear translocation of PPARγ, the main regulator of adipogenic differentiation. Fluorescence-based assays revealed increased fatty acid and disrupted low-density lipoprotein (LDL) uptake in Apilimod-treated SMCs. In Ldlr-deficient mice fed a high-fat, high-cholesterol diet for 15 weeks and treated with Apilimod for 5 weeks, we found differentially distributed plaque lipid depositions detected by Oil Red O staining.

Conclusion
Disrupting the endolysosomal maturation by PIKfyve inhibition ameliorates SMC calcification and induces a phenotypic adipogenic-like, pro-inflammatory adaption. Our data elucidates the importance of SMC phenotype monitoring during therapeutic interference. The exact lineage characteristics of these SMCs remain to be further investigated.
High pulsatile load ex vivo decreases arterial stiffness by altering the viscous, but not elastic, mechanical properties of the arterial wall
Authors: Neutel C , Wesley C , Puylaert P , De Meyer GRY , Martinet W , Guns PJ 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Foundation. Main funding source(s): Research Foundation - Flanders (Brussels, BE):
==&gt; [DUTCH]: "Fonds voor Wetenschappelijk Onderzoek (FWO) - Vlaanderen"

Introduction
The pulsatile character of blood flow in the central arteries results in a local transmission of energy to the arterial wall, acting on both the viscous and elastic components of the arterial wall. The amplitude of the "pulsatile load" is known to modulate the mechanosensitive properties of blood vessels.

Purpose
To evaluate the behavior of both the viscous and elastic components of the mouse aorta under altered pulsatile loading. Moreover, we aimed to assess regional differences in (arterial) tissue stiffness, as well as the contribution of vascular smooth muscle cell (VSMC) contractility under altered pulsatile conditions.

Methods
Aortic segments from C57Bl6 (n=9) mice were mounted in a Rodent Oscillatory Set-up for Arterial Compliance (ROTSAC), and subjected to high frequency (10 Hz) cyclic stretch at alternating loads (i.e. 20 to 100 mmHg with mean pressures ranging from 80 to 200 mmHg). Diastolic and systolic diameter, compliance, and the Peterson elastic modulus (Ep), as a measure of aortic stiffness, were determined. Viscous modulus (EV) was extracted from pressure-diameter tracings by eliminating loop hysteresis. Afterwards, the elastic modulus (EE) was calculated as the slope of the resulting pressure-diameter tracing:
(P_elastic)= (P_total) - (EV * (dD/dt))
To assess regional differences in response to changes in pulsatile load, the most proximal aortic region (i.e., thoracic ascending aorta) was compared to a more distal aortic region (i.e., abdominal infrarenal aorta).

Results
Increasing pulse pressure from 40 to 100 mmHg decreased arterial stiffness (298 ± 9 vs. 270 ± 9 mmHg; p &lt; 0.0001). Analyzing viscoelasticity revealed that increasing pulsatile load from 40 to 100 mmHg decreased EV (0.24 ± 0.02 vs. 0.21 ± 0.02 mmHg.s/mm; p &lt; 0.01) but not EE. 50 mM KCl-induced VSMC contraction increased vessel stiffness (298 ± 9 mmHg vs. 395 ± 12 mmHg; p &lt; 0.0001). This effect was pronounced at low pulse pressures (20 and 40 mmHg) while at higher pulse pressures (60, 80 and 100 mmHg) contraction-induced stiffness did not occur. Interestingly, the distal region of the aorta was stiffer than the proximal region (367 ± 60 vs. 289 ± 36 mmHg; p &lt; 0.01) and had a higher EV (0.77 ± 0.2 vs. 0.21 ± 0.05 mmHg.s/mm; p &lt; 0.0001). Increasing pulsatile load from 40 to 100 mmHg decreased the EV from distal aortic tissue more than the EV of tissue from the proximal region (-0.26 ± 0.02 vs. -0.06 ± 0.007 mmHg.s/mm).

Conclusion
High pulsatile load decreased arterial stiffness by decreasing the viscous mechanical properties. This effect was more pronounced in the distal region of the aorta.
Deciphering the role of Store Operated Ca2+ entry in post-ischemic angiogenesis
Authors: Galeano-Otero I , Del Toro R , Mayoral-Gonzalez I , Ordonez-Fernandez A , Smani T 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Agencia Estatal de Investigación

Background
Post-ischemic angiogenesis is essential to improve tissue repair and heart recovery after myocardial infarction (MI). This process is tightly controlled by pro-angiogenic factors, such as vascular endothelial growth factor (VEGF), which in turn promotes signaling pathways involving the increase in the intracellular Ca2+ concentration ([Ca2+]i). Recently, we demonstrated that store-operated calcium entry (SOCE) plays a key role in angiogenesis. However, the function of SOCE in post-ischemic angiogenesis remains unknown. The aim of this study is to explore the role of SOCE in this crucial repairing process.

Materials and methods
In vitro angiogenesis was studied using Human Umbilical Vein Endothelial Cells (HUVEC) for tube formation assay over Matrigel supplemented with serum from rat model of ischemia/reperfusion (I/R) and from ST elevation MI (STEMI) patients who underwent primary percutaneous coronary intervention (pPCI). We evaluated serum VEGF-A levels using ELISA. Genes and protein expression was determined by RT-qPCR and western blot, respectively. HUVEC migration was assessed by wound healing assay. Moreover, changes in [Ca2+]i were measured using fura-2 AM.

Results
We found that STEMI patients had higher levels of circulating VEGF-A, as compared to controls. The incubation of HUVEC seeded on Matrigel with the serum of these patients promoted vessels-like formation since it enhanced the number of meshes and junctions. Likewise, serum from I/R rats also boosted HUVEC angiogenesis over Matrigel and promoted faster gap closure in wound healing assay, as compared to cells incubated with sham rats’ serum. Furthermore, serum from both rats and patients modulated the expression Hes1 and Hey1, well-known mediators of Notch1 signaling pathway. Moreover, we found that knockdown of Orai1, the pore-forming subunit of the store-operated calcium channel (SOCC), using siRNA impairs VEGF-mediated [Ca2+]i increase, as well as HUVEC tube formation, proliferation, and migration.

Conclusion
Our data show for the first that serum from both STEMI patients and I/R rats promote angiogenesis, involving Orai1 and Notch1 signaling pathways.
Predictive value of HOMAIR indices for severity of left ventricular diastolic dysfunction and NT-proBNP activity
Authors: Sapojnic N , Caraush AP , Moscalu V , Sasu DB , Bitca AI , Durnea AS , Caraush MP 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Academy of Sciences of Moldova
Hypertension (HT) and insulin resistance (IR) often coexist and condition LV diastolic function's alteration. Diastolic dysfunction (DD) identifies hypertensives with high cardiovascular risk independent of LV mass and BP level. NT-pro BNP level serve as a diagnostic and prognostic tools in patient with DD. Several studies suggest that DD severity could be interdependent with some biomarkers (HOMAIR, NT-proBNP).

Aim
To determine if HOMA IR indices could play a predictive role in appreciating severity of LV diastolic dysfunction/ NT-proBNP activity.

Methods
101 hypertensive patients (mean age 50.11±0.79 yrs; 48.51% of men, SBP/DBP: 201.31±7.41/106.25±5.54 mmHg, BMI- 29.38±0.22 kg/m2 , HOMA IR - 2.67±0.81) with DD and IR (HOMAIR &gt; 2.5) were enrolled for the study. Ambulatory blood pressure monitoring (ABPM), transthoracic echocardiography (TE), HOMAIR and NT-pro BNP were performed at baseline and after 6, 12- months period of treatment. DD patterns were appreciated according to ASE/EACVI 2016 guideline. NT-proBNP cut-off value was considered &gt; 125 pg/ml (ESC). Correlation analysis was performed using Pearson’s test. The correlation coefficient was considered weak at &lt;0.3, medium-0.3-0.7 and strong &gt; 0.7-1.0. Statistical significance was appreciated at a P-value &lt;0.05 and highly significant at a P-value &lt;0.001.

Results
The baseline characteristics of bunch are shown in Fig 1. Diastolic dysfunction, HOMA-IR and NT-proBNP levels demonstrated a statistically significant correlation (p&lt; 0.001). A more advanced patterns of diastolic dysfunction demonstrated a particularly strong relationship with higher HOMA-IR and NT-proBNP levels ( Tab. 1).

Conclusion
In hypertensives with increase in insulin resistance, progression of the left ventricle disorder could develop. The interrelation is in such manner, that the higher is HOMAIR level, the more advanced patterns of diastolic dysfunction and NT-pro BNP activity occure.