bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 142: Transcriptome-wide Expression Quantitative Trait Loci Mapping for Atherosclerosis Susceptibility Reveals Surf4 as Novel Candidate Gene
Authors: Northoff Bernd H , Sass Kristina , Nicolaou Alexandros , Teupser Daniel , Holdt Lesca 
Published: March 2022

Previous work in an F2 intercross between atherosclerosis-susceptible C57BL/6 and atherosclerosis-resistant BALB/cByJ (BALB) mice on the LDL-receptor deficient background revealed a quantitative trait locus (QTL) for atherosclerotic lesion size at rs33142586 on chromosome (Chr) 2 in female mice (n=183) (Burkhardt et al., Arterioscler Thromb Vasc Biol; 31:775-84, 2011). The aim of the present study was to identify genetic variants responsible for differences in atherosclerosis-susceptibility between the two strains. We hypothesized that the causal genes could be identified by transcriptome-wide expression analyses followed by functional validation.
25,697 transcripts were analyzed in aortas (n=165) and livers (n=176) of F2 mice using Illumina Ref8 expression arrays. Expression levels were correlated with single nucleotide polymorphism (n=31) on Chr2 by expression QTL (eQTL) mapping. Genes co-segregating with the atherosclerosis QTL were identified as potential candidate genes. Results were validated by quantitative real-time PCR (qRT-PCR) in livers (n=176) and bone marrow-derived macrophages (BMDM, n=184) of F2 mice. Expression patterns of candidate genes were examined in different tissues of F0 mice, followed by functional studies.

eQTL mapping identified
Surf4
as the only
cis
-regulated gene co-segregating with the Chr2 QTL in aortas (LOD
array
=17.6), livers (LOD
array
=11.5, LOD
qRT-PCR
=23.2) and BMDM (LOD
qRT-PCR
=15.7).
Surf4
mRNA was significantly increased in F2 mice carrying the BALB allele at rs33142586 and BALB F0 mice. Increased
Surf4
expression was associated with reduced atherosclerosis lesion size in F2 mice. Immunohistochemical staining revealed expression of
Surf4
in vascular endothelial cells of the aorta. RNAi of
Surf4
lead to increased staurosporine- and cycloheximide-induced apoptosis and decreased proliferation compared to control siRNA in RAW 264.7 cells, which are two key mechanisms of atherosclerosis.

In conclusion, we identified
Surf4
as a novel candidate gene of atherosclerosis susceptibility co-segregating a QTL for atherosclerosis on mouse Chr2. In current work, its functional role in atherogenesis is being further investigated in
in-vitro
experiments.
Abstract 150: Trastuzumab-Associated Inflammatory Responses in Healthy Volunteers: Possible Insights in Immunogenicity of Monoclonal Antibodies
Authors: Reijers Joannes A , Moerland Matthijs , Burggraaf Jacobus 
Published: March 2022

Many monoclonal antibodies are associated with an inflammatory reaction. For example, trastuzumab induces such a response in 40-60% of treated patients, most often after the first administration. Apart from causing discomfort for the patient, these responses may increase the risk of antibody formation against the therapeutic agent. Not much is known on the pathways involved, however, comparing clinical findings and results of in vitro cytokine release between subjects who did and did not develop an inflammatory reaction may offer guidance for further investigations.
A cohort of 54 healthy male volunteers randomly received either placebo (n=8) or 6 mg/kg trastuzumab (n=46) intravenously in 90 minutes. Adverse events were captured and regular safety assessments were performed, consisting of vital signs and routine clinical laboratory tests. In addition, whole blood from a selection of volunteers with clinical signs of an inflammatory reaction as well as whole blood from trastuzumab-naïve healthy volunteers was incubated in vitro with trastuzumab.
Fever was observed in 9 (19.6%) volunteers receiving trastuzumab, occurring in combination with other influenza-like symptoms in 4. Seven subjects reached a temperature of ≥38.5°C, and four ≥39.0°C, with a maximum of 39.4°C. One volunteer developed influenza-like illness without fever. The mean latency between the onset of complaints and start of infusion was 4.7 h (SD 2.6). The fever was accompanied by a mean increase in leucocytes of 54% (SD 31), predominantly neutrophils (increase 128%, SD 60), at 24 h post administration. Although differences in baseline characteristics were not observed, a rise in neutrophils (mean increase at 24 h 94%, SD 55) and body temperature (mean peak value 37.3°C, SD 0.4) was also noted in the 37 subjects who did not develop fever. None of the placebo treated volunteers displayed this phenomenon.
These results suggest that 6 mg/kg trastuzumab is capable of inducing an inflammatory response, although the magnitude and thus the clinical manifestations differ between subjects. Data on the correlation between in vitro cytokine release, which might serve as a platform for immunogenicity research, and the observed clinical response will be presented.
Abstract 259: Inflammation and Preoxidation Are Increased in High-Density Lipoproteins From Chronic Heart Failure Subjects
Authors: Henry Sarah-Louise , Fogarty Mark , Ko Angela , Ingle Lee , Cleland John G , McEneny Jane 
Published: March 2022

Chronic heart failure (CHF) is a global term for the physiological state in which cardiac output is insufficient to meet the needs of the body and lungs. While oxidative stress, defined as an excess production of reactive oxygen species relative to antioxidant defence, has been shown to play an important role in the pathophysiology of CHF. However, although high density lipoproteins (HDL) have important cardioprotective properties, inflammation and oxidative stress can induce dramatic changes, which negate their antiatherogenic properties. Thus, such impairment to HDL may provide an important link between inflammation and oxidative stress in the pathogenesis of CHF.
This pilot study investigated the antiatherogenic properties of HDL in CHF.

Ten control subjects (BNP<100 nM) were matched for age and gender with 22 CHF subjects (BNP>100 nM). HDL
2
and HDL
3
were isolated from serum by rapid ultracentrifugation. SAA was measured by an ELISA procedure, paraoxonase-1 (PON-1) activity, hydroperoxide (HPO) and conjugated diene (CD) concentration were measured by spectrophotometric assays.

Results:
Compared to the control subjects, BNP was higher in the CHF subjects [50 (11) vs. 2439 (900) nM; p=0.015] and in HDL
2
and HDL
3
, SAA-related inflammation was increased, while the activity of the antioxidant enzyme PON-1 was decreased and CDs, one of the markers of lipid peroxidation, were increased (see table).

Conclusion:
This pilot study has shown that HDL displayed a proatherogenic phenotype in CHF subjects, which we suggest may be biologically relevant in the pathophysiology of CHF.
Abstract 338: Akt-Mediated Phosphorylation of EBP50 Regulates Skp2 Stability and Vascular Smooth Muscle Cell Proliferation
Authors: Song GyunJee , Fitzpatrick Jeremy , Pellegrini Maria , Mierke Dale F , Bisello Alessandro 
Published: March 2022

The PDZ domain-containing scaffolding protein, Ezrin-Radixin-Moesin-Binding Phosphoprotein 50 (EBP50) increases vascular smooth muscle cells (VSMC) proliferation and vascular remodeling following endoluminal vessel injury. Indeed, neointima formation is significantly reduced in EBP50 knockout mice. The mitogenic effect of EBP50 derives from the Akt-dependent stabilization of the S-phase kinase associating protein Skp2. However, the mechanisms by which Akt regulates EBP50-mediated cell proliferation are not known. We hypothesize that phosphorylation of EBP50 by Akt promotes its interaction with Skp2. This causes cytoplasmic re-localization and stabilization of Skp2 and VSMC proliferation.
Using Akt inhibitors and siRNA inhibition of Skp2, we show that the mitogenic effect of EBP50 in VSMC is dependent on both Akt and Skp2. In vitro and in vivo phosphorylation assays show that Akt phosphorylates EBP50 at T156 in the second PDZ domain. Interestingly, NMR, fluorescence polarization and co-immunoprecipitation experiments show that Skp2 interacts with the first PDZ domain of EBP50, suggesting that T156 phosphorylation exerts allosteric effects on the interaction with Skp2. Expression of EBP50 in primary VSMC induces a re-localization of Skp2 from the nucleus to the cytoplasm, leading to its stabilization and VSMC proliferation. These effects were abrogated by mutations within the first PDZ domain or of T156 to Alanine. Notably, the interaction with EBP50 does not directly affect the function of Skp2 because mutation of the C-terminal Leu to Ala, which abrogates the interaction with EBP50, does not decrease the ability of Skp2 to degrade p21cip1 and p27kip1. Consistent with the observations in primary cells, the expression of Skp2 in injured femoral arteries was reduced in EBP50 knockout mice compared to WT mice.
In conclusion, our studies identify a novel mechanism for the Akt-dependent regulation of cell proliferation by EBP50 during vascular remodeling. They indicate that inhibition of EBP50 may be useful in preventing neointima formation following angioplasty.
Abstract 377: Angiotensin II Drives Murine Ly-6chi Monocyte Trafficking from the Spleen to the Aneuvrismal Aorta Through B Cells
Authors: Mellak Safa , Loyer Xavier , Esposito Bruno , Ait Oufella Hafid , Tedgui Alain , Mallat Ziad , Potteaux Stephane 
Published: March 2022

Abdominal aortic aneurysm (AAA) is widespread among elderly people and results in progressive expansion and rupture of the aorta with high mortality. AAA is histologically characterized by medial degeneration and various degrees of chronic adventitial inflammation. In particular, macrophages initially accumulate within the site of aneurysm. Early depletion of circulating monocytes protects against the disease. Two monocyte subsets with different migratory behaviours and functions have been described in mice, ‘inflammatory’ Ly6C+ monocytes (classical) and ‘patrolling’ Ly6C- monocytes (nonclassical). In this study, we aim to understand and clarify the role of monocyte-derived cells in AAA development.
We monitored monocyte mobilisation up to 28 days in a mouse model of inflammatory AAA, which consists of implanting subcutaneous osmotic pumps delivering Angiotensin II (AngII) at a rate of 1000 ng/kg/min into C57BL6 ApoE-/- male mice between 8 and 12 weeks old. We found that AAA initiation is translated by a rapid and temporary mobilisation of classical monocytes at day 3-6 after AngII followed by non classical monocytes at days 7-9. We showed that classical monocytes are mobilized from the spleen, then recruited in the abdominal adventitia of ApoE-/- in response to AngII treatment as monitored through a pulse labelling method, along with an increased proteasic activity at the abdominal vessel wall as seen by fluorescence tomography. Importantly, we found that classical monocyte number positively correlated to stages of AAA. Splenectomy performed prior to AAA initiation prevented classical monocyte mobilization at day 3-6 after AngII and significantly protected against AAA. We found similar results in ApoE-/- Rag2-/- mice reconstituted with B cell-depleted splenocytes and treated with AngII, suggesting that splenic B cells are responsible for early deleterious mobilization of classical monocytes.
Taken together, the data show that splenic classical monocytes directly participate to AAA pathology and could be exploited to improve the prognosis of patients harbouring AAA.
Abstract 395: Identification and Characterization of a Novel Mutation in the Low-Density Lipoprotein Receptor Gene in an Omani Arab Family With Familial Hypercholesterolemia
Authors: Banerjee Yajnavalka 
Published: March 2022

Autosomal dominant familial hypercholesterolemia (FH) is caused due to mutations in the low-density lipoprotein receptor (LDLR), proprotein convertase subtilisin/kexin type 9 (PCSK9) and apolipoprotein B (ApoB) genes. Little is known regarding its prevalence in the Arab population, which not only exhibits relative genetic homogeneity, but also has a lengthy tradition of consanguinity.
In this study we screened for mutations in FH associated gene in an Omani Arab family, where FH was detected in the index patients according to the Simon Broome criteria. Effect of the identified mutations on different protein structural levels was investigated in silico.
The index patient (9-year-old female) was presented to the lipid clinic with eye xanthelasmata and thickening of both Achilles tendons, and showed off treatment total cholesterol of 896 mg/dL (23.2 mmol/L), low-density lipoprotein cholesterol (LDL-C) of 853 mg/dL (22.1 mmol/L), APOB of 4.5 g/L, triglyceride of 71 mg/dL (0.8 mmol/L), and high-density lipoprotein cholesterol of 0.74 mmol/L. She was treated with a combination of rosuvastatin/ezetimibe and LDL apheresis.
Genetic analysis of the LDLR gene showed a homozygous frame-shift deletion mutation (272delG) at exon 3, which truncates the protein after 90 amino acid residues (versus ~840 amino acids in the wild-type). The mode of inheritance of this novel mutation was also studied to construct a family tree. Additionally it was also confirmed that FH in this family is due to mutation only in the LDLR and not PCSK9 and ApoB genes. Further, the effect of the mutation was also appraised in silico on the tertiary structure of LDLR. A model of the mutant LDLR was constructed using the coordinates of the wild-type LDLR extracellular domain. Based on the model, we present a mechanistic justification behind the observed detrimental effect of the mutation on LDL-C levels.

References:
1. Al-Hinai,A.T. et al. First case report of familial hypercholesterolemia in an Omani family due to novel mutation in the low-density lipoprotein receptor gene. Angiology 64, 287-292 (2013).

2. Al-Rasadi,K. et al. Low-Density Lipoprotein Receptor Gene Mutation Analysis and Structure-Function Correlation in an Omani Arab Family With Familial Hypercholesterolemia. Angiology(2013).
Abstract 423: A Proatherosclerotic Environment Impacts the Function and Cellular Metabolism in Cardiac Mitochondria
Authors: Rocha Cynthia , Scheede-Bergdahl Celena , Whitesell Thomas , Bergdahl Andreas 
Published: March 2022

Apolipoprotein E (ApoE) is an important plasma lipoprotein, able to suppress atherosclerosis by promoting uptake of circulating cholesterol and lipid into the liver. A deficiency of ApoE (ApoE-/-) leads to fatty acid accumulation in the plasma and subsequent increase in reactive oxygen species generation. Mitochondria in cardiomyocytes, due to the nature of the tissue, rely primarily on oxidative pathways for energy production. How this altered lipid metabolism affects cardiac mitochondria, considering the heart relies predominantly on fats as its primary energy source, has yet to be elucidated. The aim of this study is to determine the influence of altered lipid metabolism, resulting from ApoE-/- on cardiac mitochondrial oxidative capacity.
We assessed the hypothesis that cardiac mitochondria would be greatly influenced by a lack of ApoE, thus affecting substrate and energy metabolism, as well as oxygen consumption compared to controls.
2-month-old ApoE-/- mice (n=8) were compared to age-matched C57Bl/6 mice (n=8). All animals were fed a standard chow and had access to water ad libitum. Left ventricular cardiac tissue samples were permeabilized and exposed to a series of titrated substrates and inhibitors that acted on the mitochondrial respiratory complexes. High-resolution respirometry was used to determine mitochondrial oxidative capacity. Citrate synthase activity, immunoblotting and immunofluorescence were also performed. Statistical comparisons were drawn using a two-tailed Student’s t-test. For statistical evaluations, p < 0.05 was considered significant.
The results revealed a decreased basal state 2 respiration, a decreased mitochondrial leak and a higher respiratory control ratio in the young ApoE-/- group compared to controls. Maximal state 3 respiration was similar in both groups.
In conclusion, these findings indicate that ApoE deficiency seems to positively affect the coupling between oxidation and phosphorylation within cardiac mitochondria in young animals. Our results suggest that ApoE-/- actually benefits cardiac metabolism, at least in the early stages of accelerated lipid metabolism. More work is needed to determine if these observations remain as the animals age or whether these effects are time dependent.
Abstract 596: Androgen Deprivation Therapy--Induced Weight Gain and Atherosclerosis in Prostate Cancer: Differences Between Orchiectomy, GnRH Agonist and Antagonist
Authors: Hopmans Sarah N , Duivenvoorden Wilhelmina C , Werstuck Geoff , Pinthus Jehonathan 
Published: March 2022

Cardiovascular disease (CVD) is one of the most common causes of mortality in prostate cancer patients. In prostate cancer treatment androgen deprivation therapy (ADT) is associated with weight gain and development of the metabolic syndrome. However, different modes of ADT can achieve castration. A
post-hoc
analysis of phase III trials suggests that GnRH antagonists are associated with less cardiac events in men with pre-existing CVD during the first year of ADT. We investigated the effects of bilateral orchiectomy, GnRH agonists and GnRH antagonists on the development of CVD in a mouse model. We hypothesize that GnRH antagonists will associate with reduced adiposity and development of atherosclerosis compared to GnRH analogues and orchiectomy.

We used LDL receptor knockout mice (n=12/group) as models for CVD to investigate and compare the effects of orchiectomy, sham surgery, sham surgery plus GnRH antagonist (degarelix) and sham surgery plus GnRH agonist (leuprolide). Longitudinal weight gain (4 months), visceral fat accumulation (CT measurements), fasting blood glucose, glucose tolerance, serum triglycerides, and testosterone levels were studied along with characteristics of aortic atherosclerotic plaques.
Leuprolide-treated mice gained significantly more weight and visceral fat compared to mice treated with degarelix. Significantly lower levels of serum triglycerides and better response to glucose loading were recorded in mice treated with degarelix. The atherosclerotic plaque area in the aortic sinus in leuprolide-treated and orchiectomized mice was significantly larger than in control mice, but not significantly different from control in degarelix-treated mice. The necrotic plaque area with degarelix was significantly smaller compared to leuprolide-treated and orchiectomized mice.
In a preclinical mouse model, the use of GnRH antagonists attenuates weight gain and development of atherosclerosis.
Abstract 642: Vascular Endothelial Growth Factor Receptor 2 Blockade in Murine Vein Graft Ameliorates Lesion Growth and Enhances Plaque Stability by Reducing Intraplaque Hemorrhage
Authors: de Vries Margreet R , de Jong Rob C , Peters Erna A , Hamming Jaap F , Goumans Marie-José , Quax Paul H 
Published: March 2022

Immature plaque neovessels contribute to atherosclerotic plaque instability and intraplaque hemorrhage by leaking erythrocytes and leukocytes in the plaque. Vascular Endothelial Growth Factor Receptor 2 (VEGFR2), together with the angiopoietin (Ang)-Tie2 system, regulates the maturation of neovessels. We have previously shown that murine vein graft lesions exhibit massive plaque neovascularization and that intraplaque hemorrhage contributes to lesion growth. We hypothesized that blockade of VEGFR2 results in more mature plaque microvessels and less intraplaque hemorrhage.
Donor caval veins were engrafted in carotid arteries of recipient hypercholesterolemic ApoE3*Leiden mice (n=14/group). Mice were treated at day 14, 17, 21 and 25 with VEGFR2 blocking antibodies (DC101) or control IgG antibodies (10 mg/kg). At day 28 mice were sacrificed for histological analysis of the vein grafts.
Morphometric analysis revealed a striking 50% decrease in vein graft segments that expressed intraplaque hemorrhage in the form of leaky vessels in the DC101 treated group. This was accompanied by a significant 25-fold decrease in extravasated erythrocytes. Furthermore, lesions that exhibit intraplaque hemorrhage showed a strong increase in Ang-2, indicative for immature neovessels. VEGFR2 blockade however, did not affect the neovessel density in the lesions (control 52±19 neovessels/section; DC101 63±25 neovessels/section). Interestingly, the vein graft lesion area in the DC101 group was significantly reduced with 32% compared to the control group. Moreover, plaque stability was clearly increased in DC101 treated mice, determined by a 25% reduction in macrophage content, a 50% increase in collagen content and a 120% increase in SMC content.
Blockade of VEGFR2 leads to reduced intraplaque hemorrhage, decreased vein graft lesion area and increased plaque stability. This identifies plaque neovascularization as an attractive target for the treatment of unstable atherosclerotic diseases.
Abstract 76: Astrocyte Tissue Factor: Double-Edged Sword of Brain Hemostasis
Authors: Wang Shaobin , Sparkenbaugh Erica M , Russell Janice , Granger D N , Mackman Nigel , Pawlinski Rafal 
Published: March 2022

Astrocytes in the brain express high levels of tissue factor (TF) presumably to prevent hemorrhage. We generated mice lacking TF gene in all neuronal cells (TFflox nestin Cre), including astrocytes. TF mRNA expression (99.5%) and total procoagulant activity (98.7%) were significantly reduced in the brains of TFflox nestin Cre mice compared to controls (TFflox). Immunostaining demonstrated the absence of TF expression on astrocytes. No spontaneous bleeding was observed in TFflox nestin Cre mice.
To investigate the role of astrocyte TF in brain hemostasis we used mouse models of ischemic and hemorrhagic stroke. Ischemic stroke was induced by 1 hour occlusion of middle cerebral artery followed by 24 hours of reperfusion. Intracerebral bleeding was observed in 72.7% of TFflox nestin Cre but not in TFflox mice subjected to ischemic stroke. Although TFflox nestin Cre mice exhibited more bleeding, they had reduced infarct size (15.8±5.4% vs 35.1±6.9%; p<0.01, mean±SD). A reduction of infarct size was also observed in mice expressing low levels of TF in all tissues (low TF mice, 18.7±6.8%) compared to wild type (WT, 29.3±6.5%; p<0.05) mice; however, no intracerebral bleeding was observed in low TF mice.
In the hemorrhagic stroke model, intracerebral administration of collagenase induced time and dose dependent bleeding in the brains of WT mice. Deletion of TF from astrocytes dramatically increases intracerebral bleeding (2.9-fold vs TFflox, p<0.01) and death (57.1%, p<0.01) 3 hours after cerebral injection of 0.025 units of collagenase. Less severe intracerebral bleeding (1.5-fold vs WT, p<0.05) and no death was observed in low TF mice. Importantly, astrocytes isolated from low TF mice had higher procoagulant activity compared to astrocytes from TFflox nestin Cre mice.
Finally, using intravital microscopy and photoactivation-induced thrombosis model, we showed that thrombosis within brain capillaries, but not arterioles or venules, was significantly attenuated (p<0.05) in TFflox nestin Cre compared to TFflox mice (consistent with direct contact of astrocyte endfeet with endothelium only within capillaries).
Our data indicate that astrocyte TF is essential for brain hemostasis. However, it also contributes to microvascular thrombosis and neuronal damage.