bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 211: Sam68 Is a Transcriptional Repressor of Endothelin-Converting Enzyme-1b Gene and Regulates Blood Pressure
Authors: Zhou Junlan , Wu Min , Cheng Min , Boriboun Chan , Biyashev Dauren , Kishore Raj , Qin Gangjian 
Published: March 2022

We have recently reported that E2F2, a transcription factor known for its role in cell proliferation and apoptosis, regulates the expression of endothelin-converting enzyme-
1b (ECE-1b) in endothelial cells and contributes to the maintenance of vascular contractility and blood pressure. However, the molecular mechanisms that underlie this novel, cell-cycle independent function of E2F2 remain largely elusive. In this current study, we profiled E2F2 interactome in the nucleus of human umbilical vein endothelial cells by utilizing a proteomic approach. Of 20 E2F2-interacting proteins identified is the Sam68, a classic RNA-binding protein known as a Src kinase substrate. Both endogenous and ectopically expressed Sam68 interact with E2F2 as confirmed by coimmunoprecipitation. Overexpression of Sam68 repressed whereas knockdown of Sam68 increased E2F2-induced ECE-1b promoter activity and mRNA expression. Chromatin immunoprecipitation analyses further confirmed that E2F2 and Sam68 colocalize on ECE-1b promoter, indicating that ECE-1b is a direct transcriptional target of E2F2 and Sam68. In vivo, Sam68 knockout (KO) mice displayed a significantly lowered blood pressure (BP) as compared to WT littermates (tail-cuff measurements of mean BP, KO vs. WT: 104 ± 6 vs. 123 ± 7 mmHg, n=25 males of age 4-6 months, P<0.05). Together, our studies have revealed a previously unknown function of Sam68 as a transcriptional co-repressor of E2F2 in the regulation of ECE-1b expression. Sam68 may link endothelial Src kinase pathways to BP regulation
Abstract 352: Unhealthy Diet and Air Pollution Induce Cardiovascular Dysfunction
Authors: Buechner Nicole , Altschmied Joachim , Ale-Agha Niloofar , Sydlik Ulrich , Jakob Sascha , Kunze Kerstin , Unfried Klaus , Haendeler Judith 
Published: March 2022

Cardiovascular diseases remain the dominant cause of deaths worldwide. Unhealthy diet and air pollution have each been described as risk factors for cardiovascular diseases. The molecular mechanisms involved in translating unhealthy diet and/or exposure to air pollution into cardiovascular dysfunctions remain yet to be identified.
First, we investigated the impact of an unhealthy diet on disease-related signaling pathways in human primary cardiovascular cells. Based on nutrition health reports on diet composition in industrialized countries, we supplemented cell culture media of human primary endothelial cells (EC), smooth muscle cells (SMC) and cardiomyocytes (CM) with 100 mg/dl LDL and replaced 1/3 of the glucose with fructose. This treatment did not induce cell death in any of the cell types. However, we observed hypertrophy in CM, enhanced proliferation in SMC and increased senescence, loss of endothelial nitric oxide synthase (eNOS) and increased nuclear localization FoxO3A in EC.
Based on several studies, which demonstrated that ultrafine particles can enter the circulation and thus directly interact with EC, we investigated the impact of ultrafine carbon black particles (ufCB), one of the major constituents of industrial and exhaust emissions, on human primary EC. Treatment with ufCB in non-toxic concentrations increased reactive oxygen species and dramatically reduced the S-NO content, a marker for NO-bioavailability. NO has been shown to increase the activation of Telomerase Reverse Transcriptase (TERT), which is required for proper endothelial cell function and is inactivated by the Src kinase under conditions of oxidative stress. ufCB significantly reduced Telomerase activity while increasing Src kinase activity. Subsequently, ufCB dramatically increased senescence of EC. To investigate whether ufCB effects also occur in vivo, we instilled ufCB in non-inflammatory concentrations into mice. Indeed, eNOS expression was reduced in the abdominal aorta of animals treated with ufCB.
Taken together, common unhealthy diet as well as low-level air pollution, which we are exposed to permanently, seem to accelerate cardiovascular diseases.
Abstract 359: Investigation of the Protein Interactions of the Shear Responsive Protein, Palladin, and Its Presence in Endothelial-Derived Microparticles
Authors: Mcginn Ciaran , Fitzpatrick Paul , Chunxu Shan , Murphy Andrew , MacDonnell Brian , Dowling Paul , Cummins Philip , Murphy Ronan 
Published: March 2022

Palladin, an actin filament associated protein, plays an important role in cell cytoskeletal regulation, with particular regards to the formation and stablilization of stress fibres. The protein is part of the myotilin-myopalladin-palladin family, but unlike myotilin and myopalladin, which only appear in striated muscle, Palladin has been observed in various cell types. The protein is encoded by the PallD gene, which through alternative splicing, results in Palladin existing in multiple different isoforms. While a number of these isoforms have been documented in cell types, it is the 90kDa isoform which is most ubiquitously expressed.
Physiological mechanotransduction such as cyclic strain or shear stress, experienced by the vasculature, has been shown to regulate expression of many genes that play an important role in the patho-physiology of cardiovascular disease (CVD), by altering cell fate and function. We have previously documented the presence of the 90kDa isoform of Palladin in Human Aortic Endothelial Cells (HAECs), along with the investigation of gene expression in response to strain and shear at different time points by utilising Western Blotting and Real Time PCR techniques.
It has been shown that gene expression increases at early time points of mechanic stimuli, followed by a reduction of expression after chronic exposure. We also observe the presence of Palladin in Endothelial Microparticles, released under inflammatory conditions. We hypothesize that, following the inflammatory response, the cell compensates for excess protein production through microparticle secretion.
Mass Spectrometric analysis of Immunoprecipited Palladin from HAECs appears to show an interaction with Drebrin (developmentally regulated brain protein), which itself binds to F-actin and is involved in cell migration. While Drebrin is mainly expressed in the brain, it has also been located in heart and muscle. We investigate here also the interactions between Palladin and Drebrin. It appears that Drebrin expression decreases as Palladin expression increases following shear stress, indicative that the two proteins compete with one another in binding to actin.
Abstract 369: Localization and Specification of a Kindlin-2 Binding Core Within Integrin Beta Cytoplasmic Tails
Authors: Bledzka Kamila , Liu Jiamin , Xu Zhen , Perera Dhanuja H , Yadav Satya P , Qin Jun , Ma Yan-Qing , Plow Edward F 
Published: March 2022

Integrins are widely expressed cell adhesion receptors. One of the key regulatory mechanisms of integrins is their ability to undergo activation. Based on in vivo studies kindlins and talin are know to play indispensable roles in integrin activation, and this function depends on their binding to the cytoplasmic tail (CT) of integrin β subunits, However, the requisite sequences within the integrin β subunit that specify kindlin is as of yet unresolved.
Using overlapping synthetic peptides corresponding to the highly conserved C-terminal region of integrin β CT as inhibitors of β3 CT: kindlin-2 interaction in pull-down assays, we identified the kindlin-2 binding core to a twelve amino-acid sequence. This sequence includes the membrane-distal NxxY region and S/T cluster, which are conserved across integrin β CT, but also variable residues flanking these two motifs. We postulated that the contribution of these flanking residues in β subunits might impose selectivity upon the interaction of kindlin-2 with various β subunits. Indeed, we found that kindlin-2 interacts with β1and β3, but poorly with the β2 CT, in both pull-down and surface plasmon resonance experiments. This selectivity was determined primarily by the non-conserved residues at the extreme C-termini of the β CT. By swapping amino acids in this region, reactivity with kindlin-2 could either be enhanced or blunted. By surface plasmon resonance, we also demonstrated that the β3 subunits can accommodate both kindlin-2 and talin head, even though their binding sites are very close in the β3 subunit. Kindlin-2 and talin head do not interact with each other but can bind simultaneously to the integrin β3 CT without mutual disturbance. This work specifies a binding site within β CT for kindlin-2, provides the first evidence for selectivity in the interaction of kindlin-2 with integrin β subunits, and excludes a spatial influence of talin and kindlin-2 on their binding to integrin CT in supporting integrin activation.
Abstract 378: Microvascular Endothelial Dysfunction and Age-Dependent Endothelial Activation in Mice with Fabry Disease
Authors: Kang Justin J , Shu Liming , Shayman James A , Bodary Peter F 
Published: March 2022

Fabry disease, a lysosomal storage disorder, leads to an age-dependent accumulation of globotriaosylceramide (Gb3) in vascular tissue that ultimately causes cardiac disease and strokes. It has been demonstrated that this accumulation of Gb3 is consistent with a decrease in nitric oxide bioavailability and an impaired endothelial function in macrovessels including the aorta. Based on this, we hypothesized that microvessels would also have vascular dysfunction in older Fabry disease mice. To evaluate this, 8 month old wild-type (WT) and alpha-galactosidase A deficient (Fabry; FA) mice (n=4 / group) were euthanized, and mesenteric artery function was tested using a pressure myograph. Vessels from WT relaxed with Ach in a dose-dependent manner, while FA vessels were nearly devoid of Ach-mediated dilatation. The maximum relaxation with the highest dose of Ach (10^-4 M) was significantly different between groups (WT: 59 ± 11 %; FA: 1 ± 12 %; p=0.0004). However, endothelium-independent relaxation mediated by sodium nitroprusside was similar in vessels from both groups (WT: 81 ± 10 %; FA: 70 ± 9 %; p=0.15), confirming that the smooth muscle layer was intact and functional. Given the similar macro- and microvascular endothelial dysfunction evident in FA, we sought to identify serum measures of endothelial activation that could be used as a surrogate determinant of the age-dependent vascular dysfunction. We collected serum from age-matched WT and FA between 2 and 17 months of age (n=4-8 / group). We observed a significant age-dependent rise in FA for several serum markers of endothelial activation including soluble e-selectin, p-selectin and vascular cell adhesion molecule. However, von willebrand factor (vWF) provided the most sensitive age- and genotype-dependent changes between WT and FA (p=0.046 at 2 months and p=0.0004 at 17 months between groups).
In conclusion, the data provide evidence of microvascular endothelial dysfunction in FA, demonstrated by a prominent defect in the mesenteric artery at 8 months of age. In addition, a robust age-dependent rise in vWF observed in FA suggests that this marker of endothelial activation might serve as a sensitive surrogate measure of endothelial dysfunction in therapeutic studies of Fabry disease.
Abstract 391: Insulin-like Growth Factors Promote Vasculogenesis in Embryonic Stem Cells
Authors: Piecewicz Stephanie M , Pandey Ambarish , Roy Bhaskar , Soh Hua Xiang , Zetter Bruce R , Sengupta Shiladitya 
Published: March 2022

The ability of embryonic stem cells to differentiate into endothelium and form functional blood vessels has been well established and can potentially be harnessed for therapeutic angiogenesis. However, after almost two decades of investigation in this field, limited knowledge exists for directing endothelial differentiation. A better understanding of the cellular mechanisms regulating vasculogenesis is required for the development of embryonic stem cell-based models and therapies. In this study, we elucidated the mechanistic role of insulin-like growth factors (IGF1 and 2) and IGF receptors (IGFR1 and 2) in endothelial differentiation using an embryonic stem cell embryoid body model. Both IGF1 and IGF2 predisposed embryonic stem to differentiate towards a mesodermal lineage, the endothelial precursor germ layer, as well as increased the generation of significantly more endothelial cells at later stages. Interestingly, differentiation into other mesoderm lineages was not augmented while endothelial cell differentiation was significantly increased. The percentage of endothelial cells within differentiated embryoid bodies was amplified by greater than 7% in embryoid bodies, a substantial increase compared to other characterized vasculogenic factors like VEGF. Inhibition of IGFR1 signaling using neutralizing antibody or a pharmacological inhibitor, picropodophyllin, significantly reduced IGF-induced mesoderm and endothelial precursor cell formation. We confirmed that IGF-IGFR1 signaling stabilizes HIF1[[Unsupported Character - Symbol Font α]] and leads to up-regulation of VEGF during vasculogenesis in embryoid bodies, which may be one of the critical mechanisms of IGF-induced promotion of vasculogenesis. Understanding the mechanisms that
are critical for vasculogenesis in various models will bring us one step closer to enabling cell based therapies for neovascularization.
Abstract 543: The Janus-Faced Transcription Factor Sister-of-Mammalian Grainyhead: 2 Isoforms with Opposing Effects in Endothelial Cells and in Vivo
Authors: Mlynek Arne , Lukosz Margarete , Köhrer Karl , Deenen René , Jakob Sascha , Güttler Christopher , Graf Martin , Winkler Christoph , Haendeler Judith , Altschmied Joachim 
Published: March 2022

Maintenance of apoptosis protection and migratory capacity are absolutely required for proper endothelial function. Several pathways are described, which are necessary for these processes including activation of the protein kinase B (Akt) and the endothelial nitric oxide synthase (eNOS). Recently, we identified a new player, the transcription factor sister-of-mammalian grainyhead (SOM)/Grainyhead-like 3, which demonstrated anti-apoptotic and pro-migratory properties in endothelial cells (EC).
In humans, three isoforms of SOM are expressed, two of which, SOM1 and SOM3, are derived from an alternatively spliced transcript. SOM3 lacks exon 2, described to contain the putative transactivation domain, and thus, has been suggested to be a repressor. SOM1 and SOM3 are co-expressed in numerous tissues, however, their exact cellular functions have not been investigated so far.
Here we demonstrate that SOM1 and SOM3 are expressed in primary human EC. In contrast to published data, which were based on a yeast two hybrid analysis with isolated domains, we could show that both full length proteins are transcriptional activators. Moreover, they have opposing functions in EC and in a whole animal model. Overexpression of SOM1 inhibited apoptosis and induced migration of EC. The latter effect could be due to the observed activation of eNOS and its upstream regulator Akt. On the contrary, SOM3 has no effect on apoptosis and significantly inhibits migration and eNOS activation. To assess the in vivo relevance of the opposing effects of SOM1 and SOM3, we made use of zebrafish as a whole animal model. SOM3, but not SOM1, induced severe malformations in embryos and reduced the number of normally developed embryos significantly. Microarray analyses showed that SOM1 and SOM3 have some overlapping, but an even larger number of distinct target genes.
Our data suggest that the opposing effects of the two SOM isoforms are mediated by transcriptome alterations and that a switch in isoform expression could change the cellular fate with dramatic consequences for a whole organism.
Abstract 93: Natural CD4+CD25+ Regulatory T Cells Control the Development and Complications of Abdominal Aortic Aneurysm in Mice
Authors: Ait-Oufella Hafid , Wang Yu , Herbin Olivier , Potteaux Stephane , Bourcier Simon , Laurans Ludivine , Esposito Bruno , Tedgui Alain , Mallat Ziad 
Published: March 2022

Abdominal aortic aneurysm (AAA) is characterized by a prominent inflammatory cell infiltrate and local destruction of extracellular matrix. Following the acute phase, the inflammatory infiltrate is predominately composed of T lymphocytes and macrophages. We have previously reported that inhibition of Th1 or Th2 cell responses had no effect on the development or the complications of experimental murine AAA. We hypothesized that another subset of T cell, natural Treg cells (Tregs), plays a role in the pathophysiology of AAA.
Male C57Bl/6 mice were infused with Angiotensin II (1μg/kg/mn) for 28 days using osmotic pumps. A control group received non-immune isotype-matched IgG antibody (n=20) and a group received anti-CD25 antibody (n=20) every 10 days that depleted more than 95% of Tregs in lymphoid organs. Treg depletion induced AAA rupture (15% vs 0%, P<0.05) and more severe AAA stages compared to control mice (P<0.01). Histological analysis showed an increase of elastin degradation (mean layer number 2.5 vs 5.6, P=0.01) and an increase of T cell accumulation in the adventitia of Treg-depleted mice (2816 vs 330 μm2, P<0.01).

CD28 or B7 deficiency leads to a profound reduction in Treg cells. We infused Angiotensin II into C57Bl/6 CD28+/+ (n=17), CD28
-/-
mice (n=11) or B7
-/-
mice (n=16). CD28 or B7 deficiency did not alter the pressor response to Angiotensin II. CD28 and B7 deficiency induced an increase of AAA incidence compared to control mice, respectively 68%, 72%, and 10%, P<0.001. Moreover, CD28 and B7 deficiency induced 20% of AAA rupture whereas no rupture was observed in control mice. In another set of experiments, transfer of Tregs (3x106 cells) into CD28
-/-
mice abolished AAA rupture (0% vs 15%, P<0.05) and significantly reduced AAA severity (P<0.01). Moreover, Treg transfer decreased elastin degradation (mean layer number 3.7 vs 2.5, P=0.04) and T cell infiltration in the adventitia (390 vs 2747 μm2, P<0.01).

Conclusion:
We have shown for the first time that natural regulatory T cell protected against AAA development and rupture.
Abstract 94: Predicting Aortic Aneurysm Rupture: A Computational Fluid Dynamics Analysis
Authors: Boyd April J , Kuhn David C , Kulbisky Gordon P , Lozowy Richard J 
Published: March 2022

Aortic size is the primary factor used to predict abdominal aortic aneurysm (AAA) rupture potential; however, this method fails to account for AAA that rupture at smaller sizes, or for those that reach extreme sizes without rupture. Currently there is no truly reliable way to evaluate the susceptibility of a particular AAA to rupture. Although computational fluid dynamic (CFD)methods have been used previously to evaluate AAA flow dynamics, these studies have yet to predict the rupture potential for specific AAA anatomy. We hypothesize that the site of maximal pressure and wall shear stress (WSS) within individual AAA will lead to biomechanical wall failure and will predict the site of aortic rupture.

In order to test this hypothesis we used commercially-available CFD software (ANSYS CFX v. 12.1) to solve the governing equations for mass and momentum for CTA-derived 3D images of ruptured AAA (RAAA) (n=5; 4 male, 1 female). Blood flow was considered to be laminar, Newtonian, and steady-state. The simulations were performed on a UNIX platform with 8 platform processors. The solution was considered to be converged when the maximum residuals for all governing equations were below 1.0 x 10
-5
. ANSYS software was used to generate a fully hexahedral mesh. Predicted intra-aortic pressure and WSS profiles were obtained.

The average AAA size at rupture was 8.3 +/- 1.52 cm. Three of the five RAAA ruptured at or near the site of maximal diameter. The maximal predicted intra-aortic pressure was 14.25 +/- 6.29 Pa and generally was localized on the anterior aortic wall. In most cases the site of actual rupture was the lateral wall of the AAA where the pressure was not significantly different from that at the site of maximal pressure (12.12 +/- 6.27 Pa, p >0.05, ns). In the normal aorta, velocity profiles are laminar to the aortic bifurcation, whereas in AAA there is significant vortex flow pattern within the aneurysm. In these RAAA the highest predicted WSS was on the anterior aortic wall and measured 0.184 +/- 0.02 Pa. At the actual site of rupture, WSS was significantly lower at 0.044 +/- 0.0006 Pa (p<0.0001). In all cases the rupture occurred in a low WSS region and was always associated with eddy formation or flow recirculation.
This CFD study was the first to model blood flow in the geometry of actual RAAA. Interestingly, the site of rupture was not in the region of maximal pressure or WSS as predicted. Rupture occurred in all cases in flow recirculation zones where low WSS predominated. This work will provide the basis for future research on a more precise prediction of rupture risk.
Abstract 120: Effects of Anacetrapib Treatment on CETP Metabolism
Authors: Reyes-Soffer Gissette , Millar John S , Lassman Michael E , Jumes Patricia , Wagner John A , Gutstein David E , Ramakrishnan Rajasekhar , Holleran Stephen , Rader Daniel J 
Published: March 2022

Background
Cholesteryl ester transfer protein (CETP) is required for the exchange of core lipids, cholesteryl esters and triglycerides (TG), between lipoproteins. This results in net exchanges of TG in very-low-density (VLDL) or low-density lipoproteins (LDL) for cholesteryl esters in high-density lipoproteins (HDL). Treatment with anacetrapib, a CETP inhibitor in phase 3 development, is associated with an increased HDL-C and apo AI, and reduced both LDL-C and triglycerides. There are no published data on the regulation of CETP levels in humans. CETP mass has been reported to increase following CETP inhibition which prompted us to examine CETP kinetics. We developed a novel method using stable isotopes and LC-MS analysis to study the effects of anacetrapib on CETP turnover.

Methods
Thirty-nine moderately hyperlipidemic participants were enrolled in a fixed-sequence study: 75% (N=29) were on atorvastatin (20mg/day) plus placebo for four weeks followed by atorvastatin plus anacetrapib (100 mg/day) for 8 weeks (S-ANA). Twenty-Five percent (N=10) of participants received double placebo for four weeks followed by placebo plus anacetrapib for 8 weeks (P-ANA). At the end of each period, we measured CETP mass, and kinetic studies were performed using D3 leucine to determine the fractional clearance and production rates of CETP.

Results
Eight weeks of anacetrapib treatment was associated with 129% increase in CETP plasma levels and an 18% decrease in CETP activity (RFU/sec) in both groups. The increase in CETP mass was associated with a significant reduction in the fractional clearance of CETP from plasma (0.48 vs. 0.23 pools/day) p <.001. The decrease in fractional clearance of CETP was similar in Panel A and Panel B (Panel A 57%, Panel B 55%). There were no significant changes in the production rate of CETP.

Conclusions
Using a novel method, we demonstrated significant effects of CETP inhibition with anacetrapib on the metabolism of CETP in plasma.