bluuebunny/crossref_metadata_2025_split huggingface dataset:
Olfactory receptor 2 signaling drives experimental abdominal aortic aneurysm formation
Authors: Schelemei P , Picard F , Nemade H , Mehrkens D , Grimm S , Tinaz K , Wagner E , Orrechioni M , Wagenhaeuser M , Schelzig H , Roy J , Liljeqvist M , Mollenhauer M , Baldus S , Winkels H 
Published: November 2023

Abstract

Background
Abdominal aortic aneurysms (AAA) are defined as enlargement above 3cm or 50% of its original diameter. Therapeutic options are limited to surgical and endovascular treatment, while no pharmacological interventions are available.
Chronic inflammation of the vessel wall and in particularly macrophage infiltration has been associated with disease progression, leading to extensive tissue remodeling, yet the underlying mechanisms are incompletely understood.
A recent study has shown extra-nasal expression of the olfactory receptor 2 (OLFR2) on vascular macrophages, in which activation induced a pro-atherogenic response.
The role of macrophage OLFR2 signaling in AAA, however, is unknown.

Methods & Results
To determine whether the OLFR2 ortholog OR6A2 is expressed in human AAA, we performed in silico micro-array analysis of human AAA tissue. OR6A2 was shown to be specifically upregulated in AAA tissue with thrombus formation compared to healthy control tissue. We confirmed OR6A2 expression on macrophages by immunofluorescence.
To determine OLFR2 expression among vascular immune cells in mice, we induced AAA formation by porcine pancreas (PPE) infusion into the infrarenal aorta of C57BL6/J (WT) mice and performed spectral cytometry utilizing a 26-marker panel at baseline, day 7 and day 28. OLFR2 expression peaked at day 7 and decreased to baseline levels at day 28 particularly on pro-inflammatory monocytes, mixed resident / migratory-like macrophages and pro-inflammatory macrophages, while OLFR2 was not detectable on tissue resident-like macrophages.
To assess a functional role of OLFR2 in AAA formation, we induced AAA in OLFR2-deficient (KO) mice and WT controls by PPE. Ultrasound analysis of the abdominal aorta revealed protection from AAA formation in KO mice. Histological analysis showed significantly reduced macrophage while increased smooth muscle cell content in aortic sections of KO mice compared to WT controls. This was accompanied by higher collagen content in KO mice. Additionally, plasma levels of the pro-inflammatory cytokines TNF-alpha and Interferon-gamma were significantly reduced in KO mice 28 days post PPE.
Bulk transcriptome analysis corroborated these findings and indicated increased pathways associated with vascular smooth muscle cell contractility while decreased leukocyte activation in KO aortas.

Conclusion
We show that OLFR2 deficiency in mice attenuates abdominal aortic aneurysm development, aortic remodeling and pro-inflammatory response. OLFR2 might thus present a novel target for drug-based AAA therapies.
Asymptomatic Brugada patients: long-term prognosis from a large prospective study
Authors: Giustetto C , Gaita F , Cerrato N , Martino A M , Millesimo M , Saglietto A , Bonacchi G , Barbonaglia L , Bianchi F , De Ferrari G M , Calo' L 
Published: November 2023

Abstract

Background
One of the greatest challenges concerning the Brugada syndrome is how to stratify and manage asymptomatic patients, who represent the majority of patients with Brugada ECG pattern (BrECG).

Purpose
Aim of our study was to evaluate the long-term prognosis of a large population of asymptomatic Brugada patients.

Methods
One thousand one hundred and forty-nine asymptomatic Brugada patients were collected. Patients were treated as follows: in the 539 subjects with spontaneous type 1 BrECG documented on basal ECG or at 12-lead 24-hour Holter monitoring (12L-Holter) an electrophysiological study (EPS) was proposed and ICD implantation was proposed on the basis of the EPS results; in the 610 subjects with drug-induced-only type 1, clinical follow-up with multiple ECGs and 12L-Holter was suggested to search for spontaneous type 1 BrECG.

Results
Over a median follow-up of 6 (4-9) years there were 17 arrhythmic events (1.5%). Sixteen events occurred in the 539 patients with spontaneous type 1 (3%; 0.4% person-years) and only 1 in the 610 patients with drug-induced type 1 BrECG (0.16%; 0.03% person-years, p 0.001). Patients with spontaneous type 1 had a 15-time higher risk of arrhythmic events than patients with drug- induced type 1 BrECG. Patients with spontaneous type 1 BrECG and positive EPS had a 3-time higher risk of arrhythmic events than patients with negative EPS (6.8% vs 1.6%, p 0.035). An ICD was implanted in 163 patients: 8 (5%) received an appropriate intervention, while 23% experienced device-related complications, including 1 death.

Conclusions
Asymptomatic patients with spontaneous type 1 BrECG have an event rate of 3% at a median follow-up of 6 years. In this group, EPS represents a useful risk stratification procedure, as a positive EPS identifies patients with a 3-time higher event rate as compared to subjects with negative EPS. A negative EPS identifies a population at low although not zero-risk. Asymptomatic patients with drug-induced only Brugada ECG pattern are a group at very low arrhythmic risk. ICD implantation is burdened by a high incidence of complications, therefore alternative therapeutic strategies are needed.
Role of S100A4 in the crosstalk between smooth muscle and inflammatory cells in atherosclerosis
Authors: Kapitanova K , Cardoso Dos Santos L M , Klingelhofer J , Azar P , Bochaton-Piallat M L 
Published: November 2023

Abstract

Background
During atherosclerotic plaque formation, smooth muscle cells (SMCs) accumulate in the intima and undergo a transition from a contractile to a synthetic phenotype. We previously showed that S100A4 is a marker of the synthetic SMCs in vitro and intimal SMCs in pig, human and mouse. In vitro, S100A4-treated SMCs acquire a pro-inflammatory phenotype. The systemic neutralization of extracellular S100A4 in ApoE-/- mice induces atherosclerotic plaque stabilization.

Purpose
To characterize the role of S100A4-expressing SMCs in atherosclerosis.

Methods and results
We established a coculture model between S100A4-stimulated human aortic SMCs and human blood-derived monocytes. Flow cytometry analysis of monocytes exposed to S100A4-treated SMCs for 3 days showed an upregulation of markers typical of inflammatory M1 (HLA-DR) and anti-inflammatory M2 (CD206) macrophages (fig. 1). Macrophages also upregulated mRNA expression of NPC Intracellular cholesterol transporter 1 and 2 by qPCR, suggesting modifications in their cholesterol metabolism. In vivo, in an SMC-lineage-tracing ApoE-/-mouse model, we identified S100A4-expressing SMCs at the onset of atherosclerosis in aortic media of 11-week-old mice fed a high cholesterol diet (HCD) for 3 weeks. Medial SMCs were characterized by means of single-cell RNA sequencing. Interestingly, 10 out of 11 identified clusters expressed S100A4, demonstrating broad and early involvement of S100A4 in shaping medial SMC identity. We assumed that blood-derived monocytes might be a driving force of early S100A4 activation in SMCs. To mitigate the impact of monocytes, 8-week-old ApoE-/- mice were fed an HCD for 9 weeks and injected with either clodronate-encapsulated or empty liposomes during the first 3 weeks. No differences in S100A4 expression in the media were observed by qPCR. Unexpectedly, ApoE-/- mice treated with clodronate demonstrated larger atherosclerotic lesions compared to control animals as measured by OilRedO staining (fig. 2). Studies are in progress to define the plaque composition.

Conclusion
Our findings shed light on the role of S100A4 in the crosstalk between SMCs and inflammatory cells in atherosclerosis and the fine-tuning of the phenotype of medial SMCs prior to their accumulation in the intima. We suggest that temporary monocyte depletion at the onset of the disease alters the mechanisms underlying plaque formation, worsening the outcome of the disease, while not affecting S100A4 expression in medial SMCs.
Myeloid cannabinoid receptor CB1 deficiency confers atheroprotection in a sex-specific manner
Authors: Wang Y , Li G , Chen B , Shakir G , Volz M , Van Der Vorst E , Maas S , Hristov M , Lacy M , Enard W , Lutz B , Weber C , Herzig S , Guillamat-Prats R , Steffens S 
Published: November 2023

Abstract

Aim
Cardiovascular diseases (CVDs) are the leading cause of death globally, claiming an estimated 17.9 million lives each year. A better understanding of the underlying chronic inflammatory disease mechanisms in atherosclerosis might help finding novel treatments and reduce CVD deaths. In particular, the cannabinoid receptor CB1 has been implicated in atherosclerosis, while its cell-specific effects in this disease are not well understood.

Methods
Myeloid Cnr1 (CB1 encoding gene) knockout mice were generated on apolipoprotein E deficiency (Apoe-/-) background by crossing CB1flox mice with transgenic mice carrying Cre under control of the lysozyme M (LysM) promoter for selective expression in myeloid cells. Age - and sex-matched groups were analyzed at baseline, 4 and 16 weeks Western diet (0,15% cholesterol). For in vitro experiments, murine bone marrow-derived macrophages (BMDM) were used.

Results
Male mice with myeloid-specific Cnr1 deficiency on atherogenic background developed smaller lesions and necrotic cores compared to controls, while only minor effects in females were observed. Male Cnr1 deficient mice had reduced arterial monocyte recruitment and macrophage proliferation with less inflammatory phenotype. The sex-specific differences were reproducible in vitro in BMDMs and blunted by female hormone estradiol treatment. Array-based kinase activity profiling of BMDMs stimulated with a synthetic CB1 agonist revealed a CB1-dependent regulation of p53 and cyclin-dependent kinases. Transcriptomic profiling via bulk RNA-sequencing of CB1 agonist-stimulated BMDMs further unveiled chromatin modifications, mRNA processing and mitochondrial respiration among the key processes affected by CB1 signaling, which was supported by metabolic flux assays. Chronic administration of the peripherally-restricted CB1 antagonist JD5037 inhibited plaque progression and macrophage proliferation, again only in male mice.

Conclusion
Impaired CB1 signaling in macrophages is atheroprotective by limiting their arterial recruitment, proliferation and inflammatory reprogramming. The biological effects of macrophage CB1 signaling seem to be more pronounced in male mice. This highlights the need to consider the biological sex as an important variable in preclinical studies.
A pH-Responsive and MRI-Functional nanomedicine enabling targeted delivery of simvastatin in atherosclerosis
Authors: Wang J W , Lin Y Z , Liu J J , Chong S Y , Kah J C Y 
Published: November 2023

Abstract

Background
Simvastatin is commonly used for the treatment of atherosclerosis. However, it causes side effects in some patients and not always effective. Moreover, although its treatment effects are often attributed to its cholesterol-lowering activity, some in vitro studies suggest that simvastatin also exerts antioxidant and anti-inflammatory effects.

Purpose
To develop an atherosclerotic plaque microenvironment-responsive nanoparticular system to deliver and release Gadolinium (Gd3+) for magnetic resonance imaging (MRI) and simvastatin for treatment within the plaque.

Methods
A nanoparticle was synthesized with nanoscale coordination polymers (NCPs), a pH-responsive linker, Gd3+, and simvastatin (ST), i.e. a ST/NCP-PEG nanoparticle. The biological effects of ST/NCP-PEG nanoparticle were tested on RAW264.7 macrophages. Four weeks old ApoE-/- mice were fed on high fat diet for 4 weeks and then randomly divided into three groups for a 8-week treatment regime via weekly intravenous administration of: 1) NCP-PEG nanoparticle without ST (control); 2) ST/NCP-PEG nanoparticles; or 3) free ST.

Results
We have developed a pH-responsive ST/NCP-PEG nanoparticle carrying an MRI contrast agent Gd3+ and a drug molecule ST, with an 80 nm diameter in spherical shape. This nanoparticle collapsed into biodegradable components while releasing both Gd3+ and ST under the atherosclerotic plaque microenvironment with a pH 5.6. ST/NCP-PEG nanoparticles exerted strong anti-oxidant and anti-inflammatory effects in vitro, as determined by an intracellular ROS indicator DCF-DA and expression levels of cytokines (e.g. IL-6, TNF-α and MCP-1), respectively. Upon intravenous administration, ST/NCP-PEG nanoparticles, as demonstrated by IVIS imaging system, accumulated specifically in the plaques in a time-dependent manner, reaching the peak concentration at 24 hours post administration. The plaques were readily visualized by MR imaging (T1-weighted signal) due to the enrichment of Gd3+. After 8 weeks treatment, compared to the empty NCP-PEG nanoparticle, treatment with ST/NCP-PEG and free ST reduced plaque lesion size by 55% (p&lt;0.001) and 29% (p&lt;0.05), respectively. Moreover, ST/NCP-PEG nanoparticles showed a larger reduction in plaque lesions than free ST (p&lt;0.05). Oil red O staining of both the whole aorta and the aortic roots generated similar results in terms of plaque lesion changes. Mechanistically, both ST/NCP-PEG and free ST reduced ROS production and increased the ratios of M2/M1 macrophages within the plaque, with ST/NCP-PEG nanoparticle treatment exerted larger therapeutic effects. Moreover, long-term treatment with ST/NCP-PEG nanoparticles did not show obvious toxicity in all major organs.

Conclusions
An atherosclerotic plaque microenvironment-responsive MR imaging functional nanoparticle for targeted delivery of ST to the plaque could diagnose atherosclerotic plaque and improve treatment efficacy of ST.
Additional effects of combination therapy of antihypertensive medication on aortic elastic properties beyond blood pressure lowering effect in spontaneously hypertensive rats
Authors: Kim S A , Koh A R , Hong D K , Suh S W 
Published: November 2023

Abstract

Background
Combination therapy in the treatment of hypertension provides better therapeutic efficacy, improved tolerability by lowering the dose and reduced potential for side effects than monotherapy. Especially, combinations of calcium channel blocker (CCB), angiotensin receptor blocker (ARB) or diuretics (thiazide) are generally effective and well tolerated regimen in antihypertensive treatment. However, we have focused more on the antihypertensive power of the combination therapy rather than the effect on the vessel itself.

Purpose
The purpose of this study is to determine the efficacy of antihypertensive medications as ARB and CCB and its combination with or without thiazide on the aortic structural and functional properties beyond the effect of blood pressure (BP) lowering.

Methods
Fourteen-week-old male spontaneous hypertensive rats were randomly divided into 5 groups, and treated with 1) vehicle (V), 2) amlodipine besylate (A), 3) olmesartan (O), 4) combined amlodipine and olmesartan (AO) or 5) combined amlodipine, olmesartan and thiazide (AOD), respectively. After 8 weeks of treatment, hemodynamic parameters and structural and functional properties of aorta as intima-media thickness (IMT) and velocity vector imaging (VVI) parameters of aorta including fractional area change (FAC), circumferential strain (CS) and strain rate (CSR) were assessed in each group and collagen contents of aorta were quantified as well.

Results
During the study period, systolic and diastolic BP were serially decreased in all treatment groups. Especially, AOD group showed lowest systolic and diastolic BP levels and also showed smallest IMT and vessel area and highest elasticity (FAC and CS) among the other groups (p&lt;0.01) at 8 weeks (Table 1). In histologic analysis, collagen contents of aorta in all treatment groups showed significant associations with BP, IMT and VVI parameters (FAC, CS, CSR) (p&lt;0.05), respectively. Particularly, aorta of AO and AOD groups showed significantly lower levels of collagen contents than that of control group. (p&lt; 0.05) (Figure 1)

Conclusion(s)
Beyond the BP lowering effect, combination of amlodipine and olmesartan with or without thiazide minimize structural changes and shows better protection for the progression of arteriosclerosis than individual monotherapy in treatment of hypertension.Table 1Figure 1
CRIP1: Role in the pathogenesis of hypertension by regulating interactions with cytoskeletal proteins
Authors: Duque Escobar J , Boateng B , Schweigert O , Riedel R , Wenzel P , Zeller T 
Published: November 2023

Abstract

Background
Hypertension is a complex polygenic disease and the underlying molecular mechanisms remain unknown. In an interdisciplinary approach, we identified cysteine-rich protein 1 (CRIP1) in human monocytes as being strongly associated with hypertension, suggesting a link between CRIP1 and the pathophysiology of hypertension via the immune system. The identification of CRIP1 protein interaction partners and associated signaling pathways would provide insight into the pathophysiological significance of CRIP1 under hypertensive conditions.

Objective
The overall aim of our project is to evaluate the functional role of CRIP1 in hypertension and cardiovascular disease (CVD). Here, using systems medicine approaches including experimental settings and human cohort data, we focused on the identification of novel CRIP1 protein interaction partners that may be associated with the pathogenesis of hypertension and cardiovascular disease.

Methods
Monocytic CRIP1 was immunoprecipitated and co-immunoprecipitated proteins in the eluate were analyzed by LC-MS/MS and Pathway and Process Enrichment Analysis and validated by immunoblot techniques. Silencing of CRIP1 was established in the monocyte-like cell line THP-1 using a vector-based shRNA system. The association of CRIP1 with genes encoding cytoskeletal proteins was accessed in a human cohort by transcriptome-based analysis of human monocytes in the Guttenberg-Gesundheitsstudie (GHS; n = 1527).

Results
First, a Yeast Two-Hybrid Assay was performed and two potential protein interaction partners were predicted for CRIP1: KTN1 and TPM2. To obtain comprehensive information on interacting proteins, a co-immunoprecipitation assay with a specific antibody against monocytic CRIP1 confirmed these as interaction partners and revealed further protein interactions with proteins that are crucial for monocyte migration and invasion behavior of monocytes by LC-MS/MS and Pathway and Process Enrichment Analysis, and immunoblot analysis. In an in vitro experiment with the human monocytic cell line THP-1, downregulation of CRIP1 by &gt;70% significantly reduce the abundance of these proteins. In addition, in a transcriptome-based analysis of human monocytes, most of the genes encoding proteins of microtubule cytoskeleton organization (GO:0000226), actin filament and cytoskeleton organization (GO:0007015 and GO:0030036), Arp2/3 complex-mediated actin nucleation (GO:0034314), and cytoskeleton-dependent intracellular transport (GO:0030705) were associated with CRIP1 expression.

Conclusion
As CRIP1 expression in human monocytes has already been linked to hypertension, our results suggest that CRIP1 may play a role in the pathogenesis of this disease by regulating interactions with cytoskeletal proteins that lead to impaired monocyte migration and invasion behaviour. Further experiments are underway to translate our findings into clinical application.
Accelerated arterial thrombus formation in Hutchinson Gilford Progeria Syndrome
Authors: Puspitasari Y M , Han J , Karch C , Ministrini S , Liberale L , Akhmedov A , Montecucco F , Beer J H , Bongiovanni D , Luscher T F , Camici G G 
Published: November 2023

Abstract

Introduction
Arterial thrombosis is an age-associated cause of myocardial infarction and stroke, which are the leading causes of mortality and morbidity worldwide. Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare genetic disorder that is caused by a point mutation in the lamin A gene. It is characterized by progressive premature aging, in which caused a decreased life expectancy. Two most frequent causes of death of children with HGPS are myocardial infarction and ischemic stroke – cardiovascular events that are tightly linked to arterial thrombosis. Despite this fact, the effect of the specific lamin A gene mutation on arterial thrombus formation remains unknown.

Methods
Transgenic heterozygous LmnaG609G knock-in (HGPS) mice and the corresponding wild-type (WT) littermate controls underwent photochemically-induced carotid artery endothelial injury to trigger thrombosis. Coagulation and fibrinolytic factors were measured in these animals. Ex-vivo platelet activation assay was also performed.

Results
HGPS mice displayed accelerated thrombus formation as underlined by shortened time to occlusion compared to WT littermates (Figure 1A-B). The level of tissue factor and von Willebrand factors are comparable between groups. No significant differences were found on factors involved in the fibrinolytic system, which was confirmed by the comparable level of D-dimers in both groups. A higher level of thrombin-antithrombin complex was observed, accompanied by lower level of antithrombin. Interestingly, HGPS animals displayed enhanced platelet activation markers expression upon ADP, collagen, and thrombin stimulation compared to WT group, demonstrating a higher platelet reactivity in progeria animals (Figure 1C).

Conclusions
Our study demonstrated an increased arterial thrombotic response in HGPS mice as compared to WT littermates which is, at least in part, mediated by higher platelet reactivity. This novel observation may provide a mechanistic explanation for the increased incidence of acute cardiovascular events observed in children with HGPS and warrant further clinical investigations also in consideration of available antiplatelet therapies.