bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cholesterol crystals as the main trigger of interleukin-6 production through innate inflammation in human spontaneously ruptured aortic plaques
Authors: Komatsu S , Yutani C , Takahashi S , Takewa M , Iwa N , Noda H , Ohara T , Kodama K 
Published: November 2023

Abstract

Aim
Cholesterol crystals (CCs) may provoke innate inflammation in experimental and animal model, however, whether CCs are bystanders or play central role for plaque rupture is unknown. This study aimed to clarify whether CCs was the main trigger of innate inflammation in human spontaneously ruptured aortic plaques (SRAPs).

Methods
This study included 260 SRAPs collected during non-obstructive general angioscopy (NOGA) from 126 patients with having or suspected CAD. Interleukin (IL)-6 levels in SRAPs were measured. IL-6 level at the Valsalva sinus, and femoral or brachial arteries were measured. IL-6 ratios, IL-6 level in SRAPs and arteries divided by IL-6 level at the Valsalva sinus at the beginning of aorta, were calculated. The number of CCs were counted in sampled blood. The correlation between the count of CCs and IL-6 levels in SRAPs and that between the counts of CCs and IL-6 ratios in SRAPs were analyzed.

Results
IL-6 ratios of the brachial and femoral arteries were 1.06 (95% CI, 0.99–1.20) and 1.11 (95% CI, 1.04–1.20), respectively. IL-6 ratio in SRAPs was 1.10 [1.00, 1.26]. (median [IQR]). CCs were detected in 94 of 260 SRAPs (36%). The count of CCs was 11,590 (95% CI: 2,386–30,113] per 10 mL in CCs positive samples. There was a moderate correlation between the counts of CCs and IL-6 ratios in SRAPs (r=0.496, r&lt;0.0001)(Fig. 1), whereas there was no correlation between the count of CCs and IL-6 levels in SRAPs.(Fig. 2)

Conclusions
By the in-situ study, CC might be the main trigger of IL-6 through innate inflammation in human SRAPs.Fig 1Fig 2
Antibody against apolipoprotein-A1, Metabolic-Associated Fatty Liver Disease and cardiovascular risk: a translational study
Authors: Pagano S , Bakker S J L , Juillard C , Vossio S , Moreau D , Brandt K J , Mach F , Dullaart R P F , Vuilleumier N 
Published: November 2023

Abstract

Background and aims
Metabolic-Associated Fatty Liver Disease (MAFLD) represents a common cause of liver disease leading to increased cardiovascular disease (CVD) morbidity and mortality. Autoantibodies against apolipoprotein A-1 (AAA-1) are a possible novel CVD risk factor promoting inflammation and disrupting intracellular lipid homeostasis, two prominent pathogenic features of MAFLD. We aimed to evaluate the possible role of AAA-1 in MAFLD and their association with CVD risk.

Methods
HepaRG cells and liver sections from ApoE-/- mice exposed to AAA-1 were used for lipid quantification and conditional protein expression. Randomly selected sera from 312 subjects of a general population cohort were used to measure AAA-1. A Fatty Liver Index (FLI) ≥60 and a 10-year Framingham Risk Score (FRS) ≥20% were used as proxy of MAFLD and high CVD risk, respectively.

Results
In-vitro and mouse models showed tha-t AAA-1 interfered with triglyceride synthesis leading to steatosis. AAA-1 generated pro-inflammatory and pro-fibrotic responses. In the 112 participants with FLI≥60, AAA-1 were associated with higher FRS, alkaline phosphatase levels, lower HDL cholesterol and tended to display higher FLI values. Univariate linear and logistic regression analyses (LR) confirmed significant associations between AAA-1, FLI and FRS≥20%, while in adjusted LR, FLI was the sole independent predictor of FRS≥20% (OR:1.05,95%CI:1.01-1.09, P=0.003) and AAA-1 was not found to be an independent FLI predictor.

Conclusion
Despite inducing a MAFLD-compatible phenotype on in vitro and in mouse models and being associated with an increased CVD risk according to FRS, the present results suggest that the AAA-1 association with CVD risk in the general population is dependent on hepatic steatosis.
Genetic Deletion of Hepatic NCOR1 regulates the bile acid metabolism and leads to reduction in atherosclerosis
Authors: Geiger M , Oppi S A , Stein S N , Costantino S , Mohammed S A , Ruschitzka F , Paneni F , Stein S 
Published: November 2023

Abstract

Background
The nuclear receptor corepressor 1 (NCOR1) plays an important role in the regulation of gene expression in immunometabolic conditions by connecting chromatin-modifying enzymes, coregulators and transcription factors. NCOR1 has been shown to be involved in cardiometabolic diseases. Since NCOR1 transrepresses the function of several nuclear receptors involved in hepatic lipid transport and synthesis, we hypothesized that genetic NCOR1 deletion in the liver alters lipid metabolism and atherosclerosis development.

Methods
To test this hypothesis, we generated hepatocyte-specific Ncor1 knockout mice on a Ldlr-/- background. Besides assessing the progression of the disease in thoraco-abdominal aortae en face, we analyzed hepatic cholesterol and bile acid metabolism at expression and functional level.

Results
This study findings demonstrated that specific Ncor1 knockout mice on an atherosclerosis-prone background develop less atherosclerotic lesions than controls. Interestingly, under chow diet plasma cholesterol levels of liver-specific Ncor1 knockout mice were slightly higher compared to control, but strongly reduced compared to control mice after feeding them an atherogenic diet for 12 weeks. Moreover, hepatic cholesterol and triglyceride contents were decreased in liver-specific Ncor1 knockout compared to control mice. Our mechanistic data revealed that NCOR1 reprograms the synthesis of bile acids towards the alternative pathway, which in turn reduce bile hydrophobicity and enhances fecal cholesterol excretion.

Conclusions
Our data suggest that hepatic Ncor1 deletion in mice decreases atherosclerosis development by reprograming bile acid metabolism and enhancing fecal cholesterol excretion. Hepatocyte NCOR1 may therefore provide a novel target for development of anti-atherosclerotic drugs.
Bile acids promote endothelial cell homeostasis via the regulation of cell metabolism and quiescence
Authors: Dietrich E , Lettlova S , Jomard A , Wasilewski G , Osto E 
Published: November 2023

Abstract

Background
Endothelial cell (EC) homeostasis is characterized by a quiescent and low-metabolic state. Obesity leads to EC dysfunction and CVD. We demonstrated that bariatric surgery improved obesity-induced EC dysfunction and increased fasting and post-prandial plasma levels of bile acids (BAs)(in particular, cholic acid (CA), chenodeoxycholic acid (CDCA) and deoxycholic acid (DCA)). BAs are emerging as metabolic signaling molecules in several tissues, but their effect on ECs has not been clearly established yet.

Purpose
We investigate the effect of BA on the maintenance of EC quiescence.

Methods
Human Aortic Endothelial Cells (HAECs) were treated for 24h with vehicle (0.1% DMSO), CA, CDCA and DCA (50uM each) in 2%FBS EGM-2 media. Percentage of apoptotic cells was assessed with AnnexinV/PI. Seahorse Mitostress test, Mitoplate and glucose uptake with 0.5µCi/ml of 14C-2-deoxy-D-glucose were used to evaluate changes of HAEC metabolism. Immunostaining for Ki67, Crystal Violet staining and scratch assay were performed to assess HAEC proliferation and migration. NO production and endothelial NO synthase (eNOS) activity were measured via DAF-2 probe and Western Blot, respectively. By qPCR genes involved in Notch pathway were evaluated. Mesoscale vascular injury panel, scratch assay, Ki67 immunostaining and glucose uptake were also performed in HAECs pre-treated with 50uM of BAs and stimulated with 0.5ng/mL TNF-α for 24h.

Results
HAECs treated with BAs increased NO production compared to vehicle and a reduction in both basal and maximal mitochondrial respiration and extracellular acidification rate was shown by Seahorse Mitostress test. Mitoplate assay displayed an overall reduction in the metabolic rate of HAECs treated with CA and DCA and a peculiar metabolic pattern in cells treated with CDCA. Reduction in glucose uptake was also observed after BA treatment, suggesting that BAs are able to decrease EC metabolism. Immunostaining for Ki67 and Crystal Violet highlighted that BAs reduced cell proliferation. The scratch assay also displayed a reduction in EC migration after BA treatment. BAs also reduced EC apoptosis, as significant differences in the percentage of AnnexinV/PI positive cells was observed in BA-treated cells compared to vehicle. qPCR showed increased expression of genes involved in Notch pathway promoting quiescence after BA treatment. Glucose uptake, scratch assay and Ki67 immunostaining performed in TNF-α stimulated HAECs suggested a BA-dependent endothelial protection in conditions of EC pro-inflammatory activation. We did not observed a reduction in pro-inflammatory biomarkers released in the supernatant of HAECs pre-treated with BA and stimulated with TNF-α.

Conclusions
Our results suggest that BAs may be involved in the maintenance of EC quiescence by regulating EC metabolism and gene expression profile. These findings highlight a potential role of BAs as endothelial-protective molecules.
Human platelets and megakaryocytes express the BPIFB4 protein and their stimulation with the recombinant Longevity-associated variant LAV-BPIFB4 displays anti-aggregating activity
Authors: Ciaglia E , Montella F , Lopardo V , Esposito M R , Carrizzo A , Vecchione C , Maciag A , Barbieri S S , Puca A A 
Published: November 2023

Abstract
Platelets are not only the main players in primary haemostasis and cardiovascular disease, but recent advances attribute them a key role in immune defence mechanisms via the release of plasma mediators of potential clinical relevance.
The bactericidal/permeability-increasing fold-containing family B, member 4 (BPIFB4) belongs to a family protein participating in the innate mechanism of host defence.
It is a secreted protein and its plasma levels inversely correlated with COVID-19 and coronary artery disease (CAD) severity and the degree of carotid stenosis and intima media thickness in human patient cohorts. Interestingly, Long Living Individuals (LLIs) displayed high BPIFB4 circulating levels and an enrichment of the 4 SNPs missense haplotype at BPIFB4 locus, the Longevity-Associated Variant (LAV). LAV-BPIFB4 genotype correlate with higher BPIFB4 plasma levels and the transfer of LAV-BPIFB4 gene delayed major age-related diseases, mainly rescuing the immunesenescence and the endothelial and cardiac dysfunctions, yet the critical assessment of the BPIFB4 protein as a platelet release product is lacking.
Herein we observed BPIFB4 release by calcium activated human platelets, a process that is amplified in LAV allele carriers donors. Furthermore, LAV gene transfer resulted in an enhanced protein enrichment in bone marrow megakaryocyte precursor cells and in a more efficient process of platelets formation in vitro. At functional level, LAV genotype was mimicked by the rhLAV-BPIFB4 platelet stimulation that induced endogenous BPIFB4 release similarly to calcium stimuli.
In a small cohort of treatment-naive patients with type 2 diabetes, while ADP and Collagene agonist stimulation induced high aggregation, the co-treatment with the rhLAV-BPIFB4 significantly reduced the activation capacity of patients’ platelets.
Accordingly, an administration of rhLAV-BPIFB4 24 hours before tromboembolism induction partly protected from mortality in a murine model of lethal pulmonary thromboembolism in vivo. As a result of reduced platelets’ activation, lower level of circulating leukocyte-platelets aggregates (LPA) was found in rhLAV-BPIFB4-treated mice compared to control.
In summary, our data report for the first time that both, human platelets and megakaryocytes, can express BPIFB4 and unravel a novel rhLAV-BPIFB4 therapeutic action in platelet iperaggregation and thrombocytopenia.
Tumor necrosis factor superfamily 14 (TNFSF14/ LIGHT) attenuates neovascularization by reducing circulating endothelial progenitor cells and function
Authors: Hsu C Y , Huang C Y , Lin S J , Lin F Y 
Published: November 2023

Abstract

Background
Tumor necrosis factor superfamily 14 (TNFSF14) is also known as LT-related inducible ligand that competes for glycoprotein D binding to herpesvirus entry mediator on T cells (LIGHT) can bind to herpesvirus invasion mediator and lymphotoxin-β receptor to perform its biological activity. LIGHT signaling strengthens the synthesis of nitric oxide, reactive oxygen species, and cytokines and affects organ function. LIGHT also stimulates angiogenesis in tumors and induces synthesis of high endothelial venules, degrades extracellular matrix in thoracic aortic dissection, induces the expression of interleukin-8, cyclooxygenase-2, and cell adhesion molecules in endothelial cells. While LIGHT induces tissue inflammation, its effects on angiogenesis after tissue ischemia are unclear. Thus, we analyzed this effect in the current study.

Methods
C57BL/6 mice underwent hind limb ischemia surgery for model construction. Doppler ultrasound, immunohistochemical staining, and western blotting were employed to analyze the effects of LIGHT on angiogenesis. In addition, human endothelial progenitor cells (EPCs) were used for in vitro studies to analyze the possible mechanisms.

Results
In the mice, LIGHT injection inhibited angiogenesis in ischemic limbs. In vitro, LIGHT inhibited the expression of integrins and E-selectin; decreased migration and tube formation capabilities, mitochondrial respiration, and succinate dehydrogenase activity; and promoted senescence in EPCs. Western blotting revealed that the impairment of EPC function by LIGHT may be due to its effects on the normal operation of intracellular Akt signaling pathway and endothelial nitrite oxide synthase (eNOS) function.

Conclusions
LIGHT inhibits angiogenesis after tissue ischemia. LIGHT inhibition of intracellular Akt-eNOS signaling axis and mitochondrial respiration in EPCs impair cell function. These events are the possible causes of impaired angiogenesis.LIGHT decreases the recovery of ischemiaSenescence and dysfunction of EPC
Impact of comorbidities on heart rate variability in patients with high cardiovascular risk
Authors: Gadaleta M , Mason F , Steinhubl S R , Quer G 
Published: November 2023

Abstract

Introduction
Heart rate variability (HRV) measures the variation in time intervals between successive heartbeats (RR intervals) and is considered a measure of a healthy regulatory system. It is influenced by a variety of factors, including age and comorbidities. We estimate temporal HRV metrics, such as the standard deviation of RR intervals (SDNN), and frequency metrics, such as the normalized high frequency component of the RR intervals (HFNU), to identify which were best for characterizing inter-individual differences by age, gender, and health status.

Purpose
Understand inter-individual differences in HRV and the influence of age, gender, and available comorbidities.

Methods
Data from at least one 2-week single-lead ECG were obtained from the 1,742 participants of the trial. Inclusion criteria for the study are defined as follows: age of 75 years or older, or a male older than age 55 years or female older than 65 years with 1 or more comorbidities that are risk factors for cardiovascular diseases. We adopted a linear regression method, where each HRV measure constitutes the outcome of a separate model, while the demographic characteristics (age and gender), the heart rate (HR), and the presence of comorbidities (yes/no for each comorbidity) are used as predictor variables. Age has been evaluated as a quadratic term to assess non-linearity of the association, and an age-gender interaction term has also been evaluated. Coefficients are expressed in terms of standard deviations, allowing for a direct comparison between metrics with different scales and variability. The HRV metrics are evaluated every 5 minutes, then the average value over a 1-day period is considered as input for the model.

Results
Temporal HRV metrics were lower among women compared with men and increased with age. They were higher in the presence of many disease states, including chronic heart failure (CHF), obesity (OBE), chronic obstructive pulmonary disease (COP), and sleep apnea (SA), but were lower among those with history of myocardial infarction (MI), chronic renal failure (CRF), and use of beta blocker medications (BB). There was no significant association of these metrics with hypertension or diabetes. For frequency metrics, analysis was limited to HFNU due to the high correlations among these measures. HFNU had strong positive associations with all co-morbidities except stroke, with particularly large coefficients for CHF, COP, and MI. HFNU values were also higher among subjects taking beta blockers. (Figure)

Conclusion
HRV can be an indicator of autonomic nervous system (ANS) dysfunction, which is often present in many diseases. By measuring HRV, clinicians can gain insights into the state of the ANS and potentially identify early warning signs of disease progression or complications.HRV and comorbidities
Immune system humanization in dyslipidemic Rag2-KO/IL2rg-KO/CD47-KO/LDL-R KO mice controls atherosclerosis
Authors: Bonacina F , Moregola A , Norata G 
Published: November 2023

Abstract

Background and aim
Given the key role of the immune response during atherosclerosis and the therapeutic interest of biologics targeting human immune cells, the need of experimental models to translate molecular mechanisms and to test therapeutic approaches for atherosclerosis is continuously increasing.
Here we describe the characteristics of an innovative immunodeficient mouse humanized with hCD34+ cells on an atheroprone background.

Methods
LDLR-KO mice were crossed with the immunodeficient C57BL/6J strain Rag2-KO/IL2rg-KO/CD47-KO (TKO, IMSR_JAX:025730) to generate an immunocompromised dyslipidemic mouse recipient TKO-LDLR KO of human hematopoietic stem cells (hCD34+).

Results
TKO-LDLR KO were first characterized for their immune and metabolic profile. TKO mice are deficient in mature lymphocytes and NK cells and this profile was conserved in TKO-LDLR KO mice. Under high cholesterol diet for 8 weeks, both male and females TKO-LDLR KO present monocytosis with increased levels of Ly6Chi monocytes compared to TKO-LDLR KO at standard diet, develop marked dyslipidemia (total cholesterol 870.9 and 890.1 mg/dL male and females respectively), steatosis and atherosclerosis. This profile confirms the suitability of TKO-LDLR KO mice for atherosclerosis studies. Next we tested the impact of immune system humanization on atherosclerosis. TKO-LDLR KO pups received a low-dose irradiation (150-200 cGy) and thereafter 1,5-2 x 10^5 hCD34+ were injected with in the liver. Engraftment of human leukocytes (hCD45+) was evaluated after two months by flow cytometry analysis from tail blood. This approach allows to reconstitute between 10-30% of hCD45+, mainly B and T cells. The humanization with hCD34+ cells affected atherosclerosis development as compared to non-humanized TKO-LDLR KO mice.

Conclusion
We have generated and characterized for the first time a humanized dyslipidemic TKO-LDLR KO mice. This mouse model presents human B and T cells and represent and important toll to investigate the impact of biologics targeted toward human targets in the context of atherosclerosis.
Growth differentiation factor 15 (GDF15) is elevated in human plaques and promotes endothelial-to-mesenchymal transition (EndMT)
Authors: Filipova V , Stuerzebecher P E , Kralisch S , Kokot K E , Kneuer J M , Erbe S , Toenjes A , Laufs U , Boeckel J N 
Published: November 2023

Abstract

Introduction and Purpose
Growth Differentiation Factor 15 (GDF15) is a member of the TGF-β superfamily and is upregulated under conditions of injury and stress such as hypoxia. GDF15 serum concentrations correlate with inflammation, cardiac fibrosis and unfavorable prognosis in cardiovascular diseases. The functional role of GDF15 in endothelial cells is largely unknown. Here, we investigate a possible role of GDF15 in the transdifferentiation of endothelial cells (EC) into mesenchymal cells (EndMT).

Methods and Results
GDF15 is expressed at mRNA and protein level in primary human ECs and is secreted as determined by qPCR, single cell sequencing (scRNA-seq) and enzyme-linked immunosorbent assay (mean ± SEM, 743.8±39.3 pg/ml, n=3). Analysis of vascular sections from Ldlr-/- mice fed a high-fat diet using spatial transcriptomics revealed increased GDF15 expression. ScRNA-seq analysis of human plaques and normal vessel sections from the same individuals revealed a 22-fold ± 5.48 (p&lt;0.05) increase in GDF15 in ECs located in plaques compared to ECs in control vessel regions. Experiments in cultured human endothelial cells showed increased GDF15 mRNA expression qPCR (+1.7-fold control, p&lt;0.05) and scRNA-seq analysis after EndMT induction.
Treatment with increasing levels of recombinant GDF15 (10, 50 and 150 ng/ml) induced EndMT as measured by expression of the EndMT marker calponin (+3.36, 7.99 and 9.53-fold, all p&lt;0.05). Interestingly, EndMT induction decreased extracellular GDF15 levels (-54.6%, p&lt;0.05), whereas intracellular levels in EC were increased (+1.9-fold control, p&lt;0.05). The use of GDF15 neutralising antibodies (1 µg/ml) reduced soluble GDF15 levels by 97.7% as measured by ELISA (p&lt;0.001). Treatment with neutralising antibodies against GDF15 reduced EndMT induction by 54% as measured by the EndMT marker calponin (p&lt;0.05).

Conclusion
In conclusion, GDF15 is elevated in mouse and human plaques, and in EndMT. Treatment with GDF15 induces EndMT, whereas inhibition by neutralising antibodies reduces EndMT induction. These data are consistent with a functional role of GDF15 in endothelial dysfunction and atherogenesis.
Hypercoagulability impairs plaque stability in diabetes-induced atherosclerosis
Authors: Ambreen S , Fatima S , Elwakiel A , Rana R , Singh K 
Published: November 2023

Abstract

Introduction
Diabetes mellitus, which is largely driven by nutritional and behavioral factors, is characterized by accelerated atherosclerosis with impaired plaque stability. Atherosclerosis and associated complications are the major cause of mortality in diabetic patients. Efficient therapeutic concepts for diabetes-associated atherosclerosis are lacking. Atherosclerosis among diabetic patients is associated with reduced endothelial thrombomodulin (TM) expression and impaired activated protein C (aPC) generation.
Methadology
Female ApoE-/- or TMPro/Pro ApoE-/- mice (age 6 to 8 weeks) were fed a normal chow diet and were made diabetic (DM) by injecting streptozotocin, reflecting type 1 DM. After 22 weeks of age, the body weight of the mice was measured, and the mice were sacrificed. Plasma and tissue samples were analysed for different parameters.

Results
Here, we demonstrate that atherosclerotic plaque stability is reduced in hyperglycemic mice expressing dysfunctional TM (TMPro/Pro mice), which have a pro-coagulant phenotype due to impaired thrombin inhibition and markedly reduced aPC generation. The vessel lumen and plaque size of atherosclerotic lesions in the truncus brachiocephalic were decreased in diabetic TMPro/Pro ApoE-/- mice compared to diabetic ApoE-/- mice. While lipid accumulation in lesions of diabetic TMPro/Pro ApoE-/- mice was lower than that in diabetic ApoE-/- mice, morphometric analyses revealed more prominent signs of instable plaques, such as a larger necrotic core area and decreased fibrous cap thickness in diabetic TMPro/Pro ApoE-/- mice. Congruently, more macrophages and fewer smooth muscle cells were observed within lesions of diabetic TMPro/Pro ApoE-/- mice.

Conclusion
Thus, impaired TM function reduces plaque stability, a characteristic of hyperglycemia-associated plaques, thus suggesting the crucial role of impaired TM function in mediating diabetes-associated atherosclerosis.