bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 255: Association of 24-Hour Heart Rate and Rate Pressure Product With Atherosclerosis Risk Factors in Nonhypertensive Individuals
Authors: Cheng Cynthia , Daskalakis Constantine 
Published: March 2022

Heart rate (HR) is an established marker of sympathetic nervous system (SNS) activity and an independent predictor of cardiovascular morbidity and mortality. However, the exact mechanism by which the SNS mediates its deleterious effects remains unknown. The main objective of this study was to determine if 24-hour mean HR and heart rate pressure product (HR24 and RPP24) obtained using ambulatory blood pressure (BP) monitoring are associated with atherosclerosis risk factors in young-middle aged non-hypertensive patients in the preclinical phase of disease.
Data were obtained from 186 non-hypertensive individuals aged 18-45. HR24 and RPP24 were significantly associated with known atherosclerosis risk factors (elevated SBP/DBP (p< 0.001), endothelial dysfunction (p< 0.05), insulin resistance (p< 0.001), and inflammatory adipokines: CRP (p< 0.001), IL-6 (p< 0.001), and PAI-1 (p< 0.001). These data suggest that elevated HR and RPP may be potential markers of elevated risk for future atherosclerotic events in these asymptomatic individuals, prior to the development of clinically defined hypertension.
Abstract 298: Effects of Long-Term Autophagy Inhibition With Chloroquine on the Development and Progression of Experimental Abdominal Aortic Aneurysms
Authors: Ramadan Azza , Wheatcroft Mark D , Quan Adrian , Singh Krishna K , Lovren Fina , Teoh Hwee , Al-Omran Mohammed , Leong-Poi Howard , Verma Subodh 
Published: March 2022

Abdominal aortic aneurysms (AAA) are a significant cause of worldwide mortality and morbidity. Although several mechanisms have been put forth, the central determinants of progressive AAA development appear to involve heightened vascular smooth muscle cell (VSMC) apoptosis, with increased degradation of the extracellular matrix, in the setting of chronic inflammation. Recent evidence suggests that autophagy genes are deregulated in human AAA and that autophagy may modulate VSMC fate and plasticity.
We sought to determine if long-term autophagy inhibition with chloroquine attenuates AAA progression and mortality in an experimental model of AAA.
ApoE-/- mice were implanted with miniosmotic pumps containing either saline (n=15) or angiotensin II (Ang II; 1.44 mg/kg/day) (n=27). Mice from each group were subdivided and administered either the potent autophagy inhibitor chloroquine (50 mg/kg/day, i.p.) or saline for 8 weeks. Hemodynamics, abdominal aortic diameter (by ultrasound) and mortality were monitored. Necroscopy evaluations, histological and immunohistological staining for markers of autophagy, apoptosis and inflammation were conducted.
Kaplan-Meier estimates were compared between the four groups over the 8-week follow-up. Ang II infusion was associated with a cumulative mortality of 38%. Long-term chloroquine treatment did not alter Ang II-associated mortality (43%, p>0.05 vs. saline-treated group). When rupture-related AAA mortality was specifically evaluated, no difference between the Ang II groups was observed. Maximal luminal diameter of the abdominal aorta was similar between the Ang II-infused groups (Saline: 1.88±0.13 mm vs. Chloroquine: 1.97± 0.17 mm, p>0.05). Ang II infusion also significantly increased MAP, an effect that was unaffected by chloroquine treatment (Saline: 140±16 mmHg vs. Chloroquine: 135±11 mmHg, p>0.05).
Long term chloroquine administration does not alter the development and/or natural history of experimental AAA. These data suggest that a global and non-specific autophagy inhibition strategy is unlikely to represent a target for intervention for AAA. Whether cell- and/or gene-specific targeting of the autophagy machinery improves AAA outcomes remains to be determined.
Abstract 333: Platelet and Endothelial Aggregation Receptor-1 Tempers Tubulogenesis via the Akt Pathway
Authors: Vandenbriele Christophe 
Published: March 2022

Platelet endothelial aggregation receptor-1 (PEAR1) is a transmembrane receptor expressed on platelets and endothelial cells (ECs). We aim to unravel its function in ECs. qPCR analyses for PEAR1 in cultured human endothelial progenitors (blood outgrowth endothelial cells (BOECs)), in ECs from human umbilical cord (HUVECs and HUAECs) and in ECs freshly isolated from multiple vascular beds showed the lowest expression in BOECs and the highest in macro- and microvascular ECs from heart and liver. HUVECs and HUAECs showed intermediate PEAR1-expression. Staining of fast growing skin hemangiomas versus normal skin ECs revealed low expression of PEAR1 in rapidly proliferating tumor ECs.
Short hairpin based miRNA lentiviral PEAR1-knockdown (shPEAR1) doubled the hybrid EA.hy926 and HUVEC EC proliferation rate compared to control lentiviral-transduced ECs. This was associated with downregulation of HES1-expression (a downstream regulator of the NOTCH-pathway), with increased baseline Akt-P levels (PI3K-activity) in shPEAR1 cells and with decreased levels of the phosphatase PTEN (PI3K-antagonist). Expression of the nuclear proliferation suppressor p21CIP1 in shPEAR1 ECs was reduced by 60±2% (mean±SEM, qPCR and Western blot), whereas the band intensity for p34/CDC2 significantly rose on Western blots (increased mitosis). PEAR1-knockdown also enhanced EC migration via reduced expression of β1- and β3-integrins both in HUVECs and EAhy926 ECs. Both enhanced proliferation and migration contributed to increased tube formation in shPEAR1 HUAECs (in vitro matrigel tube formation assays). During HUAEC tube formation, PEAR1-expression rose 5-fold, associated with a 3-fold increase of PTEN. However, PTEN-levels dropped 1,5-fold during tube formation of shPEAR1 HUAECs, compatible with increased phosphorylation levels of Akt in shPEAR1 ECs.
Our study provides the first evidence that PEAR1-expression is variable in ECs from multiple vascular beds and that PEAR1 plays a suppressive role on proliferation, migration and tube formation of ECs through regulation of the PI3K/Akt/PTEN-pathway. Our data suggest a regulatory role for PEAR1 in the NOTCH-pathway via HES1. The upregulation of PEAR1 during EC maturation is compatible with this suppressive role.
Abstract 336: The Imbalanced Redox Homeostasis in Senescent Endothelial Cells is Due to Dysregulated Thioredoxin-1 and NADPH-Oxidase 4
Authors: Altschmied Joachim , Goy Christine , Jakob Sascha , Haendeler Judith 
Published: March 2022

Senescent endothelial cells are not only present in aged vessels, but also in atherosclerotic lesions, where they contribute to endothelial dysfunction. One hallmark of cellular senescence is an imbalanced redox homeostasis. However, the underlying mechanisms how this imbalance occurs are not completely understood. Thioredoxin-1 (Trx-1) is one of the most important redox regulators in primary human endothelial cells. It has previously been shown that Trx-1, in addition to its oxidoreductase activity has anti-apoptotic properties. Cellular generators of reactive oxygen species (ROS) are NADPH oxidases (NOXs), of which NOX4 shows highest expression levels in endothelial cells. Therefore, the aim of the study was to investigate whether and how Trx-1 and NOX4 are regulated during stress-induced premature senescence in endothelial cells, an ex vivo model mirroring the situation in aged or diseased vessels.
We treated primary human endothelial cells for two weeks with hydrogen peroxide to generate stress-induced premature senescence in these cells. Senescence induction was confirmed by an increase in senescence-associated beta-Galactosidase and nuclear p21, two established senescence markers. Moreover, cytosolic and mitochondrial reactive oxygen species formation were enhanced. Concomitantly, we observed decreased Trx-1 levels and elevated NOX4 levels, which could explain the imbalance in the cellular redox homeostasis. Moreover, the lysosomal protease Cathepsin D, known to degrade Trx-1 under short-term oxidative stress, was over-activated, providing a mechanistic basis for reduced Trx-1 protein levels. Inhibition of “over-active” Cathepsin D by the specific, cell-permeable inhibitor pepstatin A counteracted the increase in nuclear p21, ROS formation and degradation of Trx-1, thus leading to blockade of stress-induced premature senscence by stabilizing the cellular redox homeostasis. Moreover, aortic Trx-1 levels negatively correlated with NOX4 expression in NOX4 transgenic mice exclusively expressing NOX4 in the endothelium and their wildtype littermates.
Our data show that loss of Trx-1 and upregulation of NOX4 may importantly contribute to the imbalanced redox homeostasis in senescent endothelial cells ex vivo and in vivo.
Abstract 428: Scavenger Receptor Class B Type I Is Required for Protection by High-Density Lipoprotein Against Doxorubicin-Induced Apoptosis in Both Mouse and Human Cardiomyocytes and Cardiotoxicity in Mice
Authors: Durham Kristina , Trigatti Bernardo 
Published: March 2022

Doxorubicin (DOX) is a clinically used anti-tumor drug that induces cardiotoxicity. HDL protects against DOX induced apoptosis in isolated cardiomyocytes, though the pathways of protection are not well known. The scavenger receptor class B type I (SRBI) is a high affinity HDL receptor, and its absence in atherogenic mice leads to occlusive CAD and MI, though a direct role for SRBI in cardiomyocytes has yet to be studied. Here we utilized both in vitro cell culture and in vivo mouse models to assess whether SRBI is required for HDL induced protection against DOX induced cardiotoxicity.
Mouse neonatal cardiomyocytes were isolated from C57Bl6 and SRBI KO hearts. SRBI was knocked down in SV40 immortalized human ventricular cardiomyocytes using siRNA. All cells were treated as follows: 24h lipoprotein starvation, 24h ±100μg/ml HDL, and 6h ±1μM DOX. Cells were then assessed for apoptosis. Secondly, male C57Bl6 or SRBI KO mice were injected i.p. with 5mg/kg DOX or saline by 5 weekly. Plasma was assessed for cardiac troponin-T. Heart sections were assessed for apoptosis and examined histologically.
HDL protected against DOX-induced apoptosis in both C57Bl6 mouse cardiomyocytes (7.7±5.0 vs 23.7±1.8% apoptotic cardiomyocytes n=3 p<.05), and in immortalized human ventricular cardiomyocytes (6.7±2.1 vs 24.5±4.8% apoptotic cardiomyocytes n=3 p<.05). Conversely, HDL was unable to protect against DOX induced apoptosis in cardiomyocytes from SRBI KO mice (37.4±6.8 vs 35.5±8.3% apoptotic cardiomyocytes, n=3 p<.05), or immortalized human cardiomyocytes transfected with siSRBI (27.2±8.9 vs 31.3±3.7% apoptotic cardiomyocytes n=3 p<.05).
In vivo studies showed that SR-BI KO hearts are more sensitive to DOX as shown by increased apoptosis (6.9±0.6 vs 3.9±0.7% apoptotic cardiomyocytes n=5/group p<.05) and greater plasma cardiac troponin T levels (1.7±0.3 vs 1.3±0.04ng/ml n=5/group p<.05). Hearts from DOX treated SRBI KO mice also exhibited greater myofibrillar loss, vacuolization, and inflammatory cell infiltration compared to C57Bl6 hearts.
Our results indicate that SRBI mediates HDL induced protection in both mouse and human cardiomyocytes. We show here that SRBI is an important factor for the protection of the heart as a whole against DOX induced cardiotoxicity.
Abstract 431: Lipoprotein Metabolism and Transendothelial Apolipoprotein A-I Transport in Mice With an Endothelium-Specific Knockout of ATP Binding Cassette Transporter A1
Authors: Hasballa Reda , Rohrer Lucia , Fotakis Panagiotis , Zannis Vassilis I , Parks John S , von Eckardstein Arnold 
Published: March 2022

Low plasma levels of high density lipoprotein (HDL) cholesterol as well as apolipoprotein A-I (ApoA-I) are associated with increased risk of coronary heart disease. HDL and ApoA-I exert multiple potentially anti-atherogenic activities. Several of them the including stimulation of cholesterol efflux from macrophage foam cells for reverse cholesterol transport are exerted in the vascular wall rather than in the plasma compartment. HDL or apoA-I must hence pass the endothelium to enter and leave the arterial wall. We previously demonstrated in vitro that the transendothelial transport of ApoA-I involves the interaction with the ATP-binding cassette transporter (ABC) A1.
To investigate the physiological relevance of transendothelial ApoA-I transport and its regulation by ABCA1, we generated an endothelium-specific ABCA1 knock-out mouse model (ABCA1e-/-) by expressing the Cre recombinase under the control of the Ve Cadherin gene in floxed ABCA1 -/- mice.
Under both chow diet and a high fat/high cholesterol diet for 16 weeks the ABCA1e-/- mice did not differ from wild type mice by plasma levels of cholesterol or triglycerides or lipoprotein profiles. After injection of radio-iodinated ApoA-I, we found no difference in the decay of plasma radioactivity and the accumulation of radiolabel in different organs except the aorta. Compared to the aortas of wild type mice, aortas of ABCA1e-/- mice showed a significantly reduced enrichment of the radiolabel. Aortas of ABCA1e-/- mice also showed a slightly decreased enrichment of radiolabeled HDL but no difference in the enrichment of radiolabeled LDL or albumin. An ApoA-I mutant with defective ABCA1 interaction showed enhanced catabolism in both wild type and ABCA1e-/- mice as compared to wild type apoA-I. However the ABCA1-defective ApoA-I mutant did not accumulate differently in the aortas of wild type and ABCA1e-/- mice.
Taken together the data support our concept and provide first in vivo evidence that transendothelial ApoA-I transport is a specific process modulated by ABCA1.
Abstract 520: Peptide and SiRNA Modulation of HSP20 Influences Vasoreactivity
Authors: Hocking Kyle M , Evans Brian , Komalavilas Padmini , Duvall Craig L , Brophy Colleen M 
Published: March 2022

Subarachnoid hemorrhages affect 30,000 people each year and accounts for between 1-7% of all strokes. Nearly 85% of the time that the hemorrhage occurs, it is due to a cerebral aneurysm that leads to delayed vasospasm and stroke. Traditional vasodilators cannot be used because they will cause systemic hypotension leaving an unmet clinical need. HSPB1 is a small heat shock related protein that modulates vasorelaxation, is downregulated in vasospastic vessels, and represents a potential therapeutic modality for the treatment of SAH-induced vasospasm.
siRNA against HSPB1 that was encapsulated with a diblock copolymer to enhance uptake, or Rat aortic smooth muscle was tested with a HSPB1 phospho-peptide (HSPB1-p) +/- a polyplex delivery system (polyplexHSPB1).
Intracellular calcium was measured concurrently with force using a Fluoroplex sysem.

Knockdown of HSPB1 inhibited sodium nitroprusside (SNP)-induced relaxation of rat aorta. In conclusion, HSP20 modulates calcium independent relaxation of vascular smooth muscle and HSPB1-p may represent a novel therapeutic for vasospasm. Tissue treated with HSPB1-p or polyplex HSPB1-p had a dose dependent inhibition of contraction. The polyplex inhibited contraction at a lower dose of HSPB1-p than peptide alone with complete inhibition of contraction at 500 μM). Complete inhibition of contraction occurred without inhibition of increases in intracellular calcium concentrations ([Ca
2+
]
i
).
Abstract 561: Human Stem Cell--Derived Models of Endothelial Cell Morphogenesis: Label-Free Analysis of Invasion, Migration and Sprouting Angiogenesis
Authors: Belair David , Carlson Coby , Abassi Yama , Irelan Jeff , Mann David 
Published: March 2022

Endothelial cell (EC) morphogenesis during angiogenesis is a highly dynamic process tightly regulated by growth factor activity in the extracellular milieu.

Current
in vitro
models of EC morphogenesis employ endpoint analyses of primary endothelial cells, such as HUVECs. These vascular cells can exhibit functional variability between donors and methods for measuring morphogenesis in response to soluble cues typically employ potentially destructive labeling and thus fail to capture the full complexity of processes involved in EC morphogenesis. Here, we describe use of human induced pluripotent stem cell derived endothelial cells (iPSC-ECs) in a real-time electronic cell sensor impedance array technology to monitor invasion and migration behavior. Additionally, an image processing algorithm was developed to assess sprouting behavior from aggregated iPSC-EC organoid cultures.

iPSC-EC responses on the impedance platform assessed the impact of serum, growth factors (
e.g.
VEGF, FGF-2), and small-molecule angiogenesis inhibitors (
e.g.
Sunitinib, SU5402) on the proliferation, migration, and invasion of iPSC- ECs compared to primary HUVECs.

Our results indicated that iPSC-ECs were more sensitive to inhibition of VEGF mediated invasion by SU5402 than HUVECs with EC50s of 4.4nM and 99nM, respectively.
In addition, matrigel invasion toward VEGF was not observed appreciably with HUVECs, whereas iPSC-ECs demonstrated a marked invasive phenotype that could be inhibited by Nocodazole at 1nM. Using a novel method to monitor the sprouting behavior of the iPSC-ECs in an aggregated organoid into matrigel by HCA and image processing, IC50s for SU5402 (2.7nM) and Nocodazole (0.4nM) were determined.
The use of iPSC-derived endothelial cells provides a robust and reproducible source of cells that outperform HUVECs. The real-time monitoring of cellular processes offers important advantages over traditional end-point assays - specifically, the ability to measure receptor activation along with morphologic and adhesive remodeling of EC upon growth factor activation and inhibition. The combination of iPSC- ECs together with real time monitoring systems provides a more biologically relevant human model system for vasculogenesis investigations.
Abstract 85: MicroRNA-210 Induces Atherosclerotic Plaque Stability Through Repression of Apoptotic Pathways in Vascular Smooth Muscle Cells
Authors: Jin Hong , Eken Suzanne , Chernogubova Ekaterina , Osterholm Cecilia , Perisic Ljubica , Razuvaev Anton , Gonzalez-Diez Maria , Roy Joy , Eriksson Per , Hedin Ulf , Maegdefessel Lars 
Published: March 2022

Aim of the present study was to explore the role of miRNAs as potential regulators in patients with carotid artery stenosis and concordant vulnerable atherosclerotic plaques.
Utilizing patient material from the Biobank of Karolinska Endarterectomies (BiKE), we were able to profile the regulation of miRNAs in patients with symptomatic vs. asymptomatic carotid stenosis undergoing carotid endarterectomy. A PCR-based miRNA array discovered 8 miRNAs (miRs-15b, -29c, -30c/d, -150, -191, -210 and -500) being substantially altered in symptomatic vs. asymptomatic patients. Next we investigated miRNA expression in 2 murine models of vascular injury and plaque vulnerability; balloon injury in Sprague-Dawley rats, and a newly-developed plaque rupture model in ApoE-/- mice, using incomplete ligation of the carotid artery with consecutive cuff injury to induce lesion rupture. In both models miR-210 was significantly repressed in response to vascular injury and plaque instability. In silico analysis revealed 48 previously validated targets of miR-210 in the vasculature. Functional miR-210 modulation analyses in both murine models revealed that the expression of three known tumorsuppressor genes (Aifm3, Apc, Casp8ap2) could be substantially altered through miR-210 modulation, affecting the proliferation rate of smooth muscle cells (SMCs) and plaque stability. By utilizing an in vitro cell streamer system, we subjected human carotid artery endothelial cells (hCAECs) and hCASMCs to laminar and oscillatory flow conditions. Interestingly, laminar flow induced miR-210 up-regulation, whereas turbulent flow suppressed miR-210 expression in both cell lines, suggesting that miR-210 repression in atherosclerotic plaques is a potential result of altered shear stress and accelerated flow patterns. Aifm3, Apc and Casp8ap2 were significantly up-regulated with disturbed flow.
The present study explores the role and therapeutic potential of miRNA regulation in patients with vulnerable atherosclerotic lesions, carotid artery stenosis and increased risk of stroke. We were able to identify miR-210 as a key modulator of pathologic processes in atherosclerosis-related vascular remodelling.
Abstract 119: Identifying and Quantifying Eight ApoA-I HDL Subspecies by a Novel Modified Sandwich ELISA
Authors: Yamamoto Rain , Furtado Jeremy D , Sacks Frank M 
Published: March 2022

HDL-cholesterol is a well-established independent risk factor used in CVD risk stratification. However, the cardioprotective effect of HDL-cholesterol is challenged by genetic studies and pharmaceutical trials, which do not support the causal relationship between increased HDL-cholesterol levels and CVD. Previously, our lab found that HDL can be classified into subspecies based on the presence of apoC-III, where for example 7% of the total plasma apoA-I in HDL has apoC-III. We also showed that the apoA-I concentration of HDL that has apoC-III predicts increased risk of CHD, whereas that of HDL that does not have apoC-III predicts decreased risk of CHD. We hypothesized that some of the many other proteins that are located in HDL are present in apoA-I HDL subspecies.
We developed a novel sandwich ELISA to measure apoA-I in HDL that is associated with a specific protein of interest, and applied it to apoA-II, apoC-III, and apoE, which define known subspecies of apoA-I HDL. We also applied the ELISA to apoC-I, apoC-II, and plasminogen, to determine whether their association with HDL constitutes a subspecies of apoA-I HDL, and in what concentrations they exist in HDL. We confirmed the existence of all the apoA-I lipoprotein subtypes using pooled plasma samples collected from healthy men and women (age: 40±14 (mean±SD), triglyceride: 116±63 mg/dl, LDL-cholesterol: 118±28 mg/dl, HDL-cholesterol: 45±10 mg/dl). The concentration (percentage) of total plasma apoA-I in HDL associated with apoA-II was 103±11 mg/dl (71±12%), with apoC-I was 23±4 mg/dl (16±2%), with apoC-II was 4±1 mg/dl (2±1%), with apoC-III was 9±3 mg/dl (6±1%), with apoE was 8±5 mg/dl (5±3 %) and with plasminogen was 4±1 mg/dl (3±1%). The results for apoA-II, apoC-III, and apoE are consistent with previously reported values. In conclusion, we established a convenient sandwich ELISA, which demonstrated the existence of new apoA-I HDL subspecies that are defined by presence of apoC-I, apoC-II, and plasminogen, as well as known subspecies of apoA-I HDL; and we quantified the concentration of such subspecies. Future studies will investigate these abundant HDL subtypes in relation to CVD risk.