bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 143: Role of Chronic Kidney Disease Promoted Caspase-1 Activation in Neointimal Hyperplasia
Authors: Ferrer Lucas M , Monroy Alexandra , Lopez Jahaira , Yang Xiao-Feng , Wang Hong , Choi Eric 
Published: July 2021

The major cause of vascular access failure is venous stenosis due to neointimal hyperplasia (NH) [62]. Vascular smooth muscle cells (VSMC) are critical for the development of NH lesions, as they have the ability to modulate their phenotype from “contractile” to a “synthetic” in the presence of uremia, through the regulation of sensor genes for uremia danger signals and VSMC-specific differentiation genes. Recent research indicates that Caspase-1 (casp-1) activation plays an essential role in sensing metabolic danger signal-associated molecular patterns[4] and initiating vascular inflammation. Therefore, the goal of this project is to examine the role of CKD-driven casp-1 activation in VSMC and CKD-related NH.
We have established a CKD mouse model and published CKD-associated vascular remodeling. We exposed wild type and caspase-1 knockout mice to our CKD model, analyzed and quantified the NH lesion formed. We also examined in vitro and ex-vivo changes in VSMC-specific differentiation genes when exposed to uremic serum and cLDL, in the presence or absence of casp-1 inhibitor.
We found that CKD serum induces with casp-1 activation and phenotypic changes in VSMCs from a “contractile” to a “synthetic” phenotype, which are reversed with casp-1 inhibition. In an ex-vivo model using relative quantification we found that VSMC contractile markers α -Actin, Calponin, SM-22, and Smoothelin gene expression of CKD mouse carotid VSMC were higher in casp-1 knockout mice when compared to wild-type (1.40, 1.28, 1.22, 1.41 respectively). Also using an in-vivo model, relative quantification of α-actin decreased from 1.0 to 0.329 when VSMCs were exposed to uremic serum and increased back to 0.588 when casp-1 inhibitor is added. The relative quantification of Calponin also decreased from 1.0 to 0.394 when exposed to uremic serum and increased back to 0.601 with caspa-1 inhibitor. We also found that casp-1 deficiency significantly reversed CKD-related vascular remodeling by reducing NH volume from 1,440,023.70 in wild-type mice to 71,069.97 μm2 in casp-1 knockouts (p-value 0.002). Our results provide a novel insight over the therapeutic potential of casp-1 inhibitors for CKD induced NH.
Abstract 15: Novel Functions of Small GTPase Rap1 in Regulating Endothelial Homeostasis: Control of Nitric Oxide Release, Vascular Function and Blood Pressure
Authors: Lakshmikanthan Sribalaji , Zheng Xiaodong , Nishijima Yoshinori , Vasquez-Vivar Jeannette , Zhang David X , Chrzanowska-Wodnicka Magdalena 
Published: July 2021

Endothelial dysfunction, resulting from decreased nitric oxide (NO) bioavailability is a pathology linked to endothelial vasomotor dysfunction and hypertension, inflammation and atherosclerosis, perturbed endothelial barrier and progression of diabetes. In blood vessels, NO is produced by the endothelial NO synthase (eNOS), the activity of which is regulated by Ca2+/calmodulin, binding of regulatory cofactors, and posttranslational modifications, including phosphorylation events on Ser1177, which stimulate NO production. Rap1 is a ubiquitously expressed small GTPase implicated in promoting vascular barrier. We have shown that endothelial cell (EC)-specific Rap1 deletion leads to defective angiogenesis in vivo due to faulty VEGFR2 activation and signaling. Importantly, EC-specific Rap1 knockout mice developed hypertension and pathological left ventricular hypertrophy.
The objective of the study was to determine the role of small G protein Rap1 in regulating endothelial NO production and endothelial-dependent vasorelaxation in vivo and ex vivo.
Using ex vivo myography and tamoxifen-inducible, endothelial-specific Rap1-knockout mice (Cadh5-CreERT2+/0;Rap1f/f), we demonstrate that Rap1 deficiency completely abrogates NO-dependent vasodilation and attenuates NO production. Mechanistically, we show that Rap1 is rapidly activated in response to receptor agonists that activate eNOS via Ca2+/calmodulin- dependent pathway and in response to shear flow, which modules eNOS activity by its phosphorylation. Rap1 deletion in human ECs, in vitro, leads to deficient NO release in response to both these stimuli, and interferes with PI3K/Akt pathway and eNOS Ser1177 phosphorylation. Further, we demonstrate Rap1 is required for transducing signals from the endothelial mechanosensing complex comprising PECAM-1, VE-cadherin and VEGFR2 in response to shear flow, leading to ligand-independent VEGFR2 activation and signaling to stimulate NO production.
We conclude that Rap1 in endothelium is critically required for endothelial homeostasis and NO production, thereby affecting vascular tone and regulation of blood pressure. Furthermore, this study establishes Rap1 as a novel regulator of mechanotransduction in response to shear flow.
Abstract 248: Hepatocyte Nuclear Factor 4α Regulates Lipoprotein Metabolism and Atherosclerosis
Authors: Xu Yang , Zalzala Munaf , Li Yuanyuan , Zhang Yanqiao 
Published: July 2021

Hepatocyte nuclear factor 4alpha (HNF4a) is a nuclear hormone receptor and highly expressed in the liver. Mutations in human HNF4a causes maturity-onset diabetes of the young type 1 (MODY1) and lipid disorder.
We investigated the role of loss of hepatic HNF4a in lipid and lipoprotein metabolism using both adenovirus-mediated gene knockdown (Ad-shHNF4a) and liver-specific HNF4a knockout (L-HNF4a-/-) mice. Our data show that loss of hepatic HNF4a caused hypolipidemia and fatty liver disease. Plasma total cholesterol and triglyceride levels were significantly reduced whereas hepatic triglyceride levels were markedly increased. There effects were caused by impaired VLDL secretion. We further determined the effect of loss of hepatic HNF4a in atherosclerosis using ApoE-/- or Ldlr-/- mice. Our data show that loss of hepatic HNF4a caused a marked reduction in the atherosclerotic lesions when mice were challenged with a Western diet. Our data indicate that hepatic HNF4a plays a critical role in modulating lipid and lipoprotein metabolism and the development of atherosclerosis. Since human HNF4a mutations are found in MODY1 patients, this study may be of clinical importance.
Abstract 252: Is Vascular Disease a Result of Too Much Vascular Damage? Evidence for an Active Endogenous Plasma Membrane Repair System in Endothelial Cells
Authors: Bernatchez Pascal-Nicolas , Sellers Stephanie 
Published: July 2021

Many studies have correlated vascular disease with excessive vascular damage. For instance, vascular endothelial cells (EC) at sites of arterial branching are exposed to turbulent flow, a highly atherogenic factor, whereas chronic hypertension accelerates endothelial dysfunction. At the molecular level, EC are routinely exposed to damaging oxidants, cholesterol and cigarette smoke chemicals. However, how the vascular endothelium ‘repairs’ after ‘damage’ was until recently unknown.
Recently, we have shown that EC express FERLINS ‘repair’ proteins, documented to allow the fusion of lipid ‘patches’ to a damaged plasma membrane. Indeed, Dysferlin and Myoferlin mRNA and proteins were found to be highly expressed in every EC line and human/rodent blood vessel investigated. Proteomics analyses revealed that both Dysferlin and Myoferlin are caveolae/lipid raft resident proteins, well known to be rich in signaling proteins at the heart of laminar flow sensing/ mechanotransduction. In vitro, loss of Dysferlin and Myoferlin resulted in attenuated Platelet Endothelial Cellular Adhesion Molecule-1 (PECAM-1) and Vascular Endothelial Growth Factor Receptor-2 (VEGFR-2) expression, respectively, as well as blunted membrane repair following physical injury. In vivo, Dysferlin-null and Myoferlin-null mice show blunted angiogenic response in response to various stimuli. Together, these data clearly show that the vasculature has complex, endogenous physical repair machinery at the plasmalemma, that the lipid ‘patches’ likely contain transmembrane protein cargo, and that Ferlins clearly play a role in vascular homeostasis.
Finally, recent preliminary data from our laboratory suggests that Dysferlin is highly overexpressed in human atherosclerotic sections, and that Dysferlin-null mice are more susceptible to cardiovascular disease that their WT counterparts, indicating a protective role for Ferlin-associated repair in the cardiovascular system.
Abstract 351: Development of an Engineered Base Cerebrovasculature Model to Study Alzheimer’s Disease in vitro
Authors: Robert Jerome , Stukas Sophie , Wellington Cheryl L 
Published: July 2021

Alzheimer disease (AD) is the most common cause of cognitive decline in the elderly, and affects more than 40% of people over 85 years of age. In addition to the defining neuropathological hallmarks of neurofibrillary tangles and parenchymal amyloid deposits within brain tissue, over 80% of AD patients also develop cerebrovascular amyloid deposits known as cerebral amyloid angiopathy (CAA). As the presence of cardiovascular and metabolic risk factors including hypertension, type II diabetes and dyslipidemia also increase AD risk, cerebrovascular function is believed to play a critical role in the etiology of AD, likely through regulating the clearance of amyloid-beta (Aβ) peptides from the brain. The cerebrovasculature has two major known mechanisms to facilitate Aβ clearance. First, Aβ interacts with the LDL receptor related protein (LRP) to directly cross the blood brain barrier (BBB). Second, Aβ can be cleared via interstitial fluid drainage pathways that form the equivalent of the lymphatic system in the central nervous system (CNS). However, a critical roadblock to progress in delineating how cerebrovascular function contributes to AD pathogenesis is the lack of model systems that recapitulate the complexity of cerebral blood vessels.
We recently engineered a novel model system using primary human endothelial cells (EC) and smooth muscle cells (SMC) cocultured under pulsatile, native-like flow condition that produces a three dimensional (3D) artery equivalent. Histological analyses showed the formation of a dense tissue composed of a tight monolayer of endothelial cells supported by a basement membrane and multiple smooth muscle cell layers. We will in a close future investigate if the engineered model developed CAA when expose to Aβ.
Abstract 545: Generation of Oxidized Phosphatidylcholines During Hypoxia-Reoxygenation Triggers Cell Death of Postnatal Cardiac Myocytes
Authors: Hasanally Devin , Margulets Victoria , Chaudhary Rakesh , Kirshenbaum Lorrie A , Ravandi Amir 
Published: July 2021

Reperfusion of ischemic heart tissue leads to a rapid burst of reactive oxygen species which initiates a cascade of events that cumulate in cell death. The most susceptible biomolecules to oxidative injury are membrane associated phospholipids. Oxidized phospholipids including oxidized phosphatidylcholines (OxPC), have shown to be potent activators of the intrinsic apoptotic signaling cascade. We hypothesize that the production of OxPC within hypoxic myocytes mediates damage from hypoxia-reoxygenation (H-R) by triggering inappropriate cardiac myocytes death. Hence, we determined whether, bioactive OxPC are produced in cardiomyocytes during H-R, and whether OxPC species induces apoptotic response resulting in cell death of cardiomyocytes.
Post-natal rat cardiomyocytes were isolated from 2 d old rat pups and submitted to primary culture in serum-free medium. Cardiac myocytes were subjected to hypoxia (H) for 1 hour in a specialized gas-tight chamber under continuous nitrogen atmosphere followed by 1 hour reoxygenation (R). Lipidomic analysis of cardiac myocytes following H-R was determined by high performance liquid chromatography linked to tandem mass spectrometry was used to quantitate 82 distinct OxPC species from cell lipid extracts. Cell viability of cardiomyocytes was assessed in the absence and presence of known OxPC molecules (POVPC, PONPC, PGPC, PAzPC) and control non-oxidized PSPC.
Lipidomic analysis of OxPC species showed increased generation of fragmented OxPC after H-R (10.20±0.67μg/mL vs1.95±0.24μg/mL, p<0.005) when compared to control cells. Cells exposed to POVPC and PONPC showed marked dose dependent increase in cell death compared to equal molar concentration of PSPC. There was also marked increase in mitochondrial permeability upon exposure to fragmented OxPC molecules.
To our knowledge this is the first demonstration that OxPC are generated within post-natal cardiomyocytes during H-R and potent inducers of cell death of cardiac myocyte during H-R. Interventions designed to modulate the synthesis of cytotoxic OxPC during H-R may prove beneficial in suppressing aberrant cardiac myocyte cell death and loss of ventricular function in patients with ischemic heart disease or following myocardial infarction.
Abstract 554: Multimerin2 Influences Vascularization Through Modulation of Perlecan
Authors: Burgisser Petra E , Tempel Dennie , Hermkens Dorien , Schulte-Merker Stefan , Cheng Caroline , Duckers Henricus J 
Published: July 2021

Vascular development is crucial in normal physiology and pathophysiology, and is in part orchestrated by a dynamic balance between extracellular matrix (ECM) and endothelial cells. The molecular mechanisms that govern this balance and as such play a role in the formation of new blood vessels remain largely unknown.
In order to identify new genes involved in blood vessel formation, we performed a genome-wide gene expression array analysis in Flk1+ angioblasts during mouse development using a commercial platform (Affymetrix). Multimerin2 (Mmrn2) expression was found to be upregulated in angioblasts and in the developing vasculature of zebrafish and mice.

In vitro
, using immunofluorescence, Mmrn2 was detected outside of human aortic endothelial cells (HAECs), were it co-localized with the extracellular matrix components perlecan, laminin and fibronectin. Direct interaction with these proteins was confirmed using a combination of co-immunoprecipitation followed by proteomics (LC-MS), and western blot to identify possible binding partners.

Mmrn2 gain-of-function and loss-of-function
in vitro
studies, using Mmrn2 siRNA and adenoviral constructs likewise decreased and increased the expression of its binding partner perlecan. Perlecan is a well known modulator of the pro-angiogenic factor, FGF2, and is required for further signaling through the FGF receptor. Therefore, Mmrn2 may influence FGF2 signaling through perlecan expression and complex formation.

Subsequent
in vitro
studies using an adenoviral construct to induce Mmrn2 overexpression resulted in a G1-arrest in HAECs (P<0.001, N=8). In FGF2 depleted medium a G1 arrest was also observed (P<0.05, N=3), Mmrn2 overexpression combined with FGF2 depleted medium did not enhance the G1 arrest, suggesting a role of Mmrn2 in FGF2 signaling.

In addition, Mmrn2 silencing or overexpression in HAECs led to a loss of ECM organization or improved organization, respectively. In line, the binding partner of Mmrn2, perlecan, has been shown to be involved in stabilization of the ECM.
These findings suggest that Mmrn2 plays a role during blood vessel formation by binding to perlecan and the ECM, thereby modulating FGF2 signaling and ECM organization and subsequently, regulating endothelial cell proliferation.
Abstract 558: Microchanneled Polysaccharide Hydrogel Laden With Endothelial Cells and Mesenchymal Stem Cells With VEGF Facilitate Formation of Vascular Structures and Are Effective for Repairing Tissue Ischemia
Authors: Lee Sangho , Lee Min Kyung , Kong Hyunjoon , Yoon Young-sup 
Published: July 2021

Various hydrogels are used to create vascular structure in vitro or to improve cell engraftment to overcome low cell survival in vivo, a main hurdle for bare cell therapy Recently we developed a modified alginate hydrogel within which microchannels are aligned to guide the direction and spatial organization of loaded cells. We investigated whether these cell constructs in which HUVECs and human mesenchymal stem cells (hMSCs) are co-loaded in this novel microchanneled hydrogel facilitate formation of vessels in vitro and in vivo, and enhance recovery of hindlimb ischemia.
We crafted a modified alginate hydrogel which has microchannels, incorporates a cell adhesion peptide RGD, and was encapsulated with VEGF. We then compared vascular structure formation between the HUVEC only (2 x 105 cells) group and the HUVEC plus hMSC group. In the HUVEC+hMSC group, we mixed HUVECs and hMSCs at the ratio of 3:1. For cell tracking, we labeled HUVECs with DiO, a green fluorescence dye. After loading cells into the microchannels of the hydrogel, these constructs were cultured for seven days and were examined by confocal microscopy. In the HUVEC only group, HUVECs stands as round shaped cells without forming tubular structures within the hydrogel. However, in the HUVEC+hMSC group, HUVECs were stretched out and connected with each other, and formed vessel-like structure following pre-designed microchannels. These results suggested that hMSCs play a critical role for vessel formation by HUVECs. We next determined their in vivo effects using a mouse hindlimb ischemia model. We found that engineered HUVEC+hMSC group showed significantly higher perfusion over 4 weeks compared to the engineered HUVEC only group or bare cell (HUVEC) group. Confocal microscopic analysis of harvested tissues showed more robust vessel formation within and outside of the cell constructs and longer term cell survival in HUVEC+hMSC group compared to the other groups.
In conclusion, this novel microchanneled alginate hydrogel facilitates aligned vessel formation of endothelial cells when combined with MSCs. This vessel-embedded hydrogel constructs consisting of HUVECs and MSCs contribute to perfusable vessel formation, prolong cell survival in vivo, and are effective for recovering limb ischemia.
Abstract 600: Asymmetric Dimethylarginine is a Novel Regulator of mTOR Expression in Adipocytes
Authors: Dowsett Laura B , Boruc Olga , Lee Ben , Leiper James 
Published: July 2021

The metabolic syndrome, in which cardiovascular disease and obesity play major roles, has become an important healthcare issue. Systemic concentrations of asymmetric dimethylarginine (ADMA) correlate with obesity and insulin resistance and it is considered to be an independent marker of cardiovascular disease. Currently, ADMA is understood to be an endogenous inhibitor of nitric oxide synthase; however, the direct effect of ADMA on adipocytes and in obesity has not been investigated. Here, we establish that ADMA in adipocytes regulates the expression and activity of the mTOR lipid synthesis pathway which results in adipocyte hypertrophy both in vitro and in vivo.
Treatment of 3T3-L1 adipocytes with ADMA (10μM) resulted in adipocyte hypertrophy (control 27μm2 ±1.4, 10μM ADMA 33μm2± 1.6). This could not be mimicked through the blockade of NO by L-NAME (28μm2 ± 1.5) and PB-ITU (25μm2± 1.2). Hypertrophy was driven by an increase in mTOR expression (x2.4 ± 0.4) which upregulates ACC (x4.6 ± 0.8) and FASN (x1.4 ± 0.1) suggesting ADMA increases lipid production in an NO-independent manner.
Investigations in vivo were explored through the genetic manipulation of DDAH1, an enzyme responsible for ADMA breakdown. Specific deletion of DDAH1 within adipocytes increased intracellular ADMA concentrations. This in turn caused an upregulation of mTOR, ACC and FASN expression and resulted in adipocyte hypertrophy. Conversely, genetic overexpression of DDAH1 reduced ADMA production within adipocytes and caused a down-regulation in mTOR expression.
These data suggests that ADMA is a novel regulator of mTOR expression and activity at physiological concentrations. Pathological concentrations of ADMA are associated with adipocyte hypertrophy a known risk factor for insulin resistance and inflammation.
Abstract 665: Activation of Histone Deacetylase 2: A Novel Strategy for Reversing Vascular Dysfunction in Atherogenesis
Authors: Pandey Deepesh , Sikka Gautam , Bergman Yehudis , Kim Jae H , Ryoo Sungwoo , Romer Lewis , Berkowitz Dan 
Published: July 2021

Arginase 2 is a critical target in atherosclerosis as it regulates both endothelial NO, fibrosis and inflammation. The increase in Arg2 activity with endothelial cell activation is dependent on both early post-translational dependent mechanisms as well as a later increase in Arg 2 expression. The regulators of Arg2 transcription in the endothelium have not been characterized. The goal of current study is to determine the role of specific HDACs in the regulation of endothelial Arg2 transcription and thereby endothelial function.
The global HDAC inhibitor, trichostatin (TSA) both time and concentration-dependently increased Arg2 mRNA, protein levels and activity in both HAECs and mouse aortic rings, a process that leads to Arg2-dependent endothelial dysfunction. TSA and atherogenic stimulus enhances activity of common promoter regions of Arg 2. All non-selective Class I HDAC inhibitors (TSA, Scriptaid, varinostat) enhanced Arg2 expression, while only the, the HDCA 1 and 2 selective inhibitor, mocetinostat (MGCD) enhances Arg2 expression. Overexpression of HDAC 2, 3 or 8 in HAECs have no effect on Arg 2 expression while HDAC2 cDNA overexpression concentration-dependently suppresses Arg2 expression. Conversely, siRNA knockdown of HDAC2 enhances Arg2 expression. Additionally like TSA, mouse aortic rings pre-incubated with MGCD resulted in endothelial dysfunction. Finally HDAC inhibition with TSA decreased endothelial NO and increased ROS production in an arginase-inhibitable manner.
In conclusion, HDAC2 is critical regulator of Arg2 expression thereby regulating endothelial NO and ROS production, and consequently endothelial function. Overexpression or activation of HDAC2 thus represents a novel therapy for the prevention and treatment of endothelial dysfunction and atherosclerosis.