bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 53: Ablation of Thrombin Signaling in Platelets but Not in Endothelial Cells Impairs Hemostasis in a Mouse Model of Spontaneous Gastrointestinal Bleeding
Authors: Wilson Stephen J , Stevens Maria M , Coughlin Shaun R
Published: July 2021

Human PAR1 is expressed in endothelial cells as well as in platelets where it facilitates the response to thrombin and platelet activation. Vorapaxar, a PAR1 antagonist, prevents myocardial infarction and stroke in patients with prior MI or peripheral arterial disease at a cost of increased bleeding risk. Par1 is also highly expressed in endothelial cells in mice, and Par1-deficiency is associated with bleeding in the mouse embryo at midgestation. Additionally, known actions of endothelial PAR1 activation suggest pro-hemostatic functions. This raises the question of whether inhibition of PAR1 function in endothelial cells (in addition to PAR1 inhibition in platelets) contributes to the bleeding risk associated with Vorapaxar treatment. Our previous work demonstrated that Par1 deficiency results in loss of thrombin signaling in mouse endothelial cells but not mouse platelets, while Par4 deficiency ablated thrombin-induced platelet activation in mice. Thus, mice allow us to separate loss of thrombin signaling in platelets from loss of thrombin signaling in endothelial cells. Accordingly, we used Apc min/+ mice, which develop intestinal polyposis and spontaneous GI bleeding, as a model to determine whether loss of thrombin signaling in platelets (Par4 KO) or endothelial and other cells (Par1 KO) exacerbates spontaneous bleeding. Hematocrit and other hematologic parameters were measured biweekly from 7 weeks through 15 weeks of age. Hematocrits in mice wild-type for Apc were stable over this period (41.48 ± 0.48 at 7 weeks; 40.48 ± 0.37 at 15 weeks, n=15). Hematocrits in Apc min/+ mice fell approximately linearly from 37.06 ± 0.82 at 7 weeks to 14.39 ± 1.12 at 15 weeks (n=15). Hematocrits in Par1-deficient Apc min/+ mice were indistinguishable from those in Apc min/+ without Par deficiency (14.39 ± 1.12 vs 14.47 ± 1.66 at 15 weeks; n=6-15). By contrast, Par4-deficient Apc min/+ mice were already severely anemic at 7 weeks compared to Apc min/+ mice (19 ± 2.0 vs 39 ± 3.6; p<0.01, n=4). Par-dependent differences in polyp count and size were not detected. Taken together, our results suggest that loss of thrombin signaling in platelets promotes spontaneous GI bleeding in the Apc min model while loss of thrombin signaling in endothelial cells is without effect in this system.
Abstract 536: Poor Correlation Between FMD and PAT-Ratio in Patients With Metabolic Syndrome
Authors: Tomohiro Ueda
Published: July 2021

Background:
Flow mediated dilatation (FMD) and peripheral arterial tonometry (PAT) are commonly used methods for assessing endothelial function in a research setting but it is unclear how well they correlate. The aim of this study is to compare and correlate these methods in patients with metabolic syndrome.

Methods:
The study involved 105 subjects (mean age68±10 years) with metabolic syndrome. Based on the results of coronary angiography, they were divided into 2 groups: a study group with coronary lesions (n=68) and a control group without coronary lesions (n=37). Flow mediated vasodilatation (FMD) and nitroglycerine-induced vasodilatation (NID) in the brachial artery was measured by using UNEXEF18G (UNEX CO, Japan). At the same time, PAT ratio was measured by using Endo-PAT 2000 (Itamar Medical, Israel)

Results:
FMD was not correlated with PAT ratio by Spearman`s analysis. FMD was significantly impaired in the study group compared to that in the control group (3.9±1.8% vs. 2.6±1.5%, respectively; P<0.001). However, NID and PAT ratio had no deference in the two groups. Multivariable analysis revealed that FMD (odds ratio: 0.53, 95% confidence interval [CI]: 0.31-0.90) were independent variables for CAD in metabolic syndrome patients.

Conclusion:
This study showed that poor correlation between FMD and PAT in patients with metabolic syndrome. PAT may not be used as a substitute for FMD as a measure of endothelial function
Abstract 545: Cinnamon Extract Inhibit Lipids and Inflammation by Modulation of Transcription Factors:
in vivo
Model
Authors: Tuzcu Zeynep , Orhan Cemal , Sahin Nurhan , Sahin Kazim , Juturu Vijaya
Published: July 2021

Objectives:
CNM (CNM) polyphenol has antioxidant and anti-inflammatory properties. Therefore, we hypothesized CNM decrease heart disease risk factors and may enhance anti- inflammatory properties in rats fed a diet containing CNM. In this study, we evaluated the effects of CNM polyphenol on insulin resistance (IR), hyperlipidemia, hepatic transcription factors expressions [sterol regulatory element-binding protein1c (SREBP-1c), liver X receptor-α (LXR-α), nuclear factor kappa B p65 (NF-κB p65), nuclear factor-E2-related factor-2 (Nrf2)] in rats fed high fat diet (HFD).

Method:
Twenty-eight Wistar rats were allocated into four groups; (i) normal control; animals fed with normal chow (C ) (ii) CNM (C+CNM 100 mg/kg b.wt.), (iii) HFD (42% of calories as fat, high fat diet [HFD]), and (iv) HFD + CNM for 12 weeks. Blood analysis for triglycerides (TG) and cholesterol (CHOL), glucose, insulin, malonaldehyde (MDA a marker of oxidative stress [OS]) were estimated. Body weight, visceral fat and liver weight recorded and liver MDA assessed. SREBP-1c, LXR-α, ATP-citrate lyase (ACLY), fatty acid synthase (FAS), NF-κB p65 expressions and decreased the PPAR-α, p-IRS-1, Nrf2, HO-1 proteins were evaluated by Western blotting.

Results:
HFD rats of the liver had increased SREBP-1c, LXR-α, ATP-citrate lyase (ACLY), fatty acid synthase (FAS), NF-κB p65 expressions and decreased the PPAR-α, p-IRS-1, Nrf2, HO-1 expressions compared to control group. CNM supplementation decreased body weight (8.4%), visceral fat (36.6%), liver weight (17.7%), serum glucose and insulin concentrations, lipid profile (
P
< 0.05), and serum and liver MDA (23.3% and 25.4 %) concentration compared to HFD rats (
P
< 0.05). CNM decreased hepatic SREBP-1c (18.1 %), LXRα (27.9%), ATP-citrate lyase (ACLY, 22.7 %), fatty acid synthase (FAS, 15.8 %), NF-κB p65 (23.3%) and enhanced the PPAR-α, IRS-1 (72.7 %), Nrf2 (111.7 %) and HO-1 (72.1 %) proteins in HFD rat livers (
P
< 0.05).

Discussion:
These results suggest CNM supplementation reduces hyperlipidemia, inflammation, and oxidative stress through activating transcription factors (SREBP-1c, LXR-α, NF-κB, and Nrf2) and anti-oxidative defense signaling pathway.
Abstract 560: Ideal Cardiovascular Health and Subclinical Markers of Carotid Structure and Function the Paris Prospective Study III
Authors: Gaye Bamba , Mustafic Hazrije , Laurent Stéphane , Perier Marie-Cécile , Thomas Frédérique , Tafflet Muriel , Pannier Bruno , Boutouyrie Pierre , Jouven Xavier , Empana Jean-Philippe
Published: July 2021

Objective:
We hypothesized that subclinical markers of vascular structure and function, which are independent predictors of cardiovascular disease, would be less frequent in subjects with ideal than poor cardiovascular health (CVH) as defined by the American Heart Association (AHA).

Approach and Results:
Carotid parameters were measured using high-precision echotracking device in 9155 nonreferred participants attending a health checkup in a large health center in Paris (France) between 2008 and 2012. According to the AHA, participants with 0 to 2, 3 to 4, and 5 to 7 metrics (smoking, physical activity, body mass index, diet, blood glucose and total cholesterol, blood pressure) at the ideal level were categorized as having poor, intermediate, and ideal CVH. Carotid parameters were dichotomized according to their median value, and multivariable logistic regression analysis was performed. Mean age was 59.55 (SD 6.3) years; 39% were females, and ideal CVH was present in 10.11% of the study participants. After adjustment for age, sex, education, and living alone and compared with a poor CVH, an ideal CVH was associated with lower common carotid artery intima–media thickness (odds ratio=1.64; 95% confidence interval 1.40, 1.93), absence of carotid plaques (odds ratio=2.14; 95% confidence interval 1.60, 2.87), lower Young’s elastic modulus (odds ratio=2.43; 95% confidence interval 2.07, 2.84), and higher carotid distensibility coefficient (odds ratio=2.90; 95% confidence interval 2.47, 3.41).

Conclusions:
In community subjects aged 50 to 75 years, ideal CVH was associated with substantially less arterial stiffness and thickness. These associations might contribute to the lower risk of cardiovascular diseases in subjects with ideal CVH.
Abstract 562: Human Carotid Atherosclerosis Plaque Progression and Vessel Material Stiffness at Follow Up Had Positive Correlation: An in vivo Vessel Stiffness MRI Follow Up Study
Authors: Wang Qingyu , Tang Dalin , Canton Gador , Guo Jian , Guo Xiaoya , Hatsukami Thomas S , Billiar Kristen L , Wu Zheyang , Yuan Chun
Published: July 2021

It is hypothesized that artery stiffness may be associated with plaque progression. However, in vivo vessel material stiffness follow-up data is lacking in the literature.

In vivo 3D multi-contrast and Cine magnetic resonance imaging (MRI) carotid plaque data were acquired from 8 patients with follow-up (18 months) with written informed consent obtained. Cine MRI and 3D thin-layer models were used to determine parameter values of the Mooney-Rivlin models for the 81slices from 16 plaques (2 scans/patient) using our established iterative procedures. Effective Young’s Modulus (YM) values for stretch ratio [1.0,1.3] were calculated for each slice for analysis.

Stress-stretch ratio curves from Mooney-Rivlin models for the 16 plaques and 81 slices are given in Fig. 1. Average YM value of the 81 slices was 411kPa. Slice YM values varied from 70 kPa (softest) to 1284 kPa (stiffest), a 1734% difference. Average slice YM values by vessel varied from 109 kPa (softest) to 922 kPa (stiffest), a 746% difference. Location-wise, the maximum slice YM variation rate within a vessel was 306% (139 kPa vs. 564 kPa). Average slice YM variation rate within a vessel for the 16 vessels was 134%. Average variation of YM values from baseline (T1) to follow up (T2) for all patients was 61.0%. The range of the variation of YM values was [-28.4%, 215%]. For progression study, YM increase (YMI=YM
T2
-TM
T1
) showed negative correlation with plaque progression measured by wall thickness increase (WTI), (r= -0.6802, p=0.0634). YM
T2
showed strong negative correlation with WTI (r= -0.7764, p=0.0235). Correlation between YM
T1
and WTI was not significant (r= -0.4353, p= 0.2811).

Conclusion
In vivo carotid vessel material properties have large variations from patient to patient, along the vessel segment within a patient, and from baseline to follow up. Use of patient-specific, location specific and time-specific material properties could potentially improve the accuracy of model stress/strain calculations.
Abstract 571: Short Term Taste Preference for Cholesterol Rich Diet in ABCA1 Null Mice
Authors: Tsujita Maki , Ugawa Shinya , Yokoyama Shinji
Published: July 2021

Background:
Plasma HDL particle is one of the essential player for maintaining cholesterol homeostasis in mammal. Even though cholesterol can be ubiquitously biosynthesized upon requirement, cholesterol in diet is efficiently used to maintain cholesterol homeostasis which implicates existence of sensing system for cholesterol in diet. The study of taste preference on cholesterol was not well explored. In this project we examined taste preference in mice using a novel method called dual diet test (DDT) system.

Methods:
Chow pellets of 1.2% cholesterol or 0.02% Acesulfame potassium (AmK, calorie-free sweet molecule) were prepared based on standard mouse chow, CE-2(0.2% cholesterol, Crea Japan). The mouse chow tray was divided into three sections. Water nozzle was set in the center section. Each chow was weighed then set ether right or left section. One session was composed by 1-3 days and remaining chow was weighed to calculate consumption of each chow diet. The diet position was switched randomly at each session. The operation was performed between ZT9-ZT10 when mice were less active in a day. To avoid social isolation stress, 2-5 littermates were placed per cage.

Results:
C56B/6 wild type mice and LCAT null mice significantly preferred 1.2% cholesterol containing chow compared to control chow during 6 sessions by 18-28 weeks old littermates (n=16, n=8, respectively). These mice also significantly preferred AmK-chow compared to the control chow. ABCA1 null and ABCA1 heterozygote mice exhibited significance on the preference of 1.2% cholesterol in the first 3 sessions but not for the later sessions. Repeating the experiment after interval period improved their cholesterol preference in ABCA1 null mice. No gender difference was observed. Preferences to AmK-chow were significant in all trials.

Conclusions:
The preference in higher cholesterol or sweet in diet were detected at this novel DDT system, significantly. In ABCA1 null or heterozygote mice, long-term preferences to high cholesterol diet were abolished perhaps due to their lack of focus on cholesterol-rich chow of which less reward to these mice. In this experiment, mice synchronized in consuming higher cholesterol containing diet indicating mice are sensing cholesterol as a preferred tastant.
Abstract 575: ApoE Determines the Functional Fate of ApoC-III: Hepatic Clearance vs. Lipase Activity
Authors: Ramms Bastian , Nora Chelsea , Al-assam Norah , Graham Mark , Gordts Philip
Published: July 2021

Clearance of triglyceride (TG)-rich lipoproteins (TRLs) is mediated by three main receptors syndecan-1 (SDC1), low-density lipoprotein receptor (LDLR) and LDLR-related protein 1 (LRP1). Previous studies revealed that interactions of apolipoprotein (apo) E and apoA-V on TRLs with heparan sulfate chains of SDC1 are required for TLR clearance. ApoC-III is another apolipoprotein and emerging factor in regulating TG metabolism. We recently showed that apoC-III inhibits hepatic TRL clearance through the LDLR/LRP1 axis and that apoC-III accumulates on TRLs in mutant mice lacking functional SDC1 (
Ndst1
f/f
Alb-Cre
+
), suggesting that apoC-III enriched TRLs are preferentially cleared by SDC1. In this study we determined the impact of apoE, a common ligand for all three receptors, on apoC-III metabolism. In
Apoe
-/-
Ndst1
f/f
Alb-Cre
+
mice antisense oligonucleotide (ASO) lowering of apoC-III reduced plasma TG levels. In contrast to
Ndst1
f/f
Alb-Cre
+
mice the apoC-III ASO was not associated with improved hepatic TRL clearance in [
3
H]retinol excursion studies in
Apoe
-/-
Ndst1
f/f
Alb-Cre
+
mice. Binding and uptake experiments in primary hepatocytes with apoE-deficient TRLs were not affected by the presence or absence of apoC-III. Reconstitution of the same TRLs with apoE inhibited clearance only of apoC-III enriched TRLs indicating that apoE is essential for apoC-III-mediated inhibition of TRL clearance. Further analysis revealed that apoC-III ASO improved lipase activity in
Apoe
-/-
Ndst1
f/f
Alb-Cre
+
mice. It remains to be determined if apoC-III lowering in the absence of apoE stimulates lipoprotein lipase or hepatic lipase or both. In conclusion we demonstrate that apoC-III blocks TRL clearance in an apoE-dependent manner and inhibits lipase activity in the absence of functional apoE.
Abstract 576: Salicylamine, A γ-ketoaldehyde Scavenger, Improves HDL Function and Reduces Atherosclerosis in Apoe Deficient Mice
Authors: Huang Jiansheng , Yancey Patricia , Ding Lei , Zhang Youmin , Oates John , Amarnath Venkataraman , Roberts Jackson , Davies Sean , Linton MacRae F
Published: July 2021

Background:
Lipid peroxidation products impair the cholesterol efflux capacity of high-density lipoprotein (HDL) and promote the development of atherosclerosis. The impact of inhibition of malondialdehyde (MDA)-HDL adduct formation by scavengers on HDL function and whether small molecule aldehyde scavengers protect against the development of atherosclerosis was examined.

Methods and Results:
Western blot analysis of ApoAI revealed that the amount of ApoAI crosslinking increased with MDA concentration. In the presence of LPS, MDA-HDL (HDL modified by 1mM MDA) versus control HDL stimulated 2- and 1.8-fold more expression of TNF-α and IL-1β in
Apoe-/-
macrophages demonstrating that MDA-HDL has reduced anti-inflammatory function. HDL-mediated macrophage cholesterol efflux was decreased by ~ 42%, 55%, 70%, and 80%, respectively, for HDL modified with 0.125 mM, 0.25 mM, 0.5 mM, and 1mM MDA, demonstrating that MDA modification of HDL affects its cholesterol efflux capacity in a dose dependent manner. Analysis by Western blot demonstrated that 5mM of salicylamine (SAM) and 5mM of pentylpyridoxamine (PPM), γ-ketoaldehyde scavengers, attenuated MDA mediated crosslinking of apoA-I in HDL (molar ratio of MDA and HDL is 1:5) by 60% and 80 % (P<0.05), respectively. Both SAM and PPM maintained the cholesterol efflux capacity of MDA treated HDL in
Apoe-/-
macrophages. In addition, pretreatment of LDL with SAM prevented MDA-ApoB adduct formation, and compared to incubation with LDL containing MDA-ApoB adducts, SAM treatment resulted in 57% less cholesterol accumulation in J774 macrophages. Importantly, administration of the ketoaldehyde scavenger, SAM, versus the nonreactive analogue, 4-SAM, to
Apoe-/-
mice consuming a Western diet for 16 weeks reduced the extent of proximal aortic atherosclerosis by 28% (P<0.05).

Conclusions:
Treatment with salicylamine, a γ-ketoaldehyde scavenger: 1) inhibits MDA-ApoA1 adduct formation thereby preserving HDL cholesterol efflux capacity; 2) prevents MDA-apoB100 formation resulting in less macrophage cholesterol accumulation; 3) reduces atherosclerosis in

Apoe
-/-

mice. These results support the therapeutic potential of salicylamine in the treatment of atherosclerotic cardiovascular disease.
Abstract 590: Autophagy is Required for Endothelial Atheroprotective Signaling Under Physiological Flow
Authors: Kheloufi Marouane , Vion Anne-Clemence , Hammoutene Adel , Poisson Johanne , Lasselin Juliette , Devue Cecile , Pic Isabelle , Dupont Nicolas , Busse Johanna , Stark Konstantin , Loyer Xavier , Souyri Michele , Viollet Benoit , Julia Pierre , Tedgui Alain , Codogno Patrice , Boulanger Chantal M. , Rautou Pierre-Emmanuel
Published: July 2021

Objective:
Preferential development of atherosclerotic lesions in areas of low shear stress is associated with increased endothelial inflammation, apoptosis and senescence. On the contrary, high shear stress areas are protected from plaque development, but the mechanisms remain elusive. Autophagy is a protective mechanism allowing recycling of defective organelles and proteins to maintain cellular homeostasis. Our aim was to understand the role of autophagy in athero-protective effects of high shear stress.

Approach and Results:
We used the parallel plate chamber system in vitro to generate different shear stress conditions on endothelial cell deficient or not in autophagy (shRNA Atg5 vs. control) and examined autophagic flux in cells transfected with RFP-GFP-LC3 plasmid. Endothelial autophagy was silenced in

Atg5
flox/flox
; VE-cadherin-cre

mice. We first demonstrated in human and murine arteries and in cultured endothelial cells that atheroprotective shear stress activates endothelial autophagic flux. On the opposite, endothelial cells exposed to atheroprone low shear stress displayed inefficient autophagy, associated with activation of mTOR, inhibition of AMPKα pathways and blockade of the autophagic flux. Interestingly, plaque burden augmented specifically in areas usually atheroresistant in

ApoE
-/-

hypercholesterolemic mice deficiency in endothelial autophagy. Both in cultured endothelial cells and in transgenic mice, deficiency in endothelial autophagy was associated with a defect in endothelial alignment with flow direction, a hallmark of endothelial cell health. Deficiency in endothelial autophagy increased endothelial senescence and TNFα-induced inflammation under high shear stress conditions. These effects were associated with impaired KLF2 activation. In transgenic mice, endothelial senescence and apoptosis was augmented in high shear stress areas of the descending thoracic aorta when compared to control littermate mice.

Conclusions:
Altogether, these results show that adequate endothelial autophagic flux under high shear stress limits atherosclerotic plaque formation by preventing inflammation, senescence and apoptosis.
Abstract 606: Reciprocal Gene-Gene Regulation Between GWAS Genes
ADTRP
and
MIA3
Contributing to Susceptibility of Coronary Artery Disease
Authors: Wang Fan , Luo Chunyan , Ren Xiang , Ke Tie , Xu Chengqi , Qin Subo , Tang Bo , Yao Yufeng , Chen Qiuyun , Wang Qing K
Published: July 2021

Genome-wide association studies (GWAS) have identified >60 genomic loci for coronary artery disease (CAD), including
ADTRP
(regulator of TFPI and coagulation) and
MIA3
(involved in ER trafficking of collagens). In this study, we hypothesized that some GWAS genes form a molecular regulatory network involved in the pathogenesis of CAD. Global microarray analysis showed that
ADTRP
regulated expression of
MIA3
and
PIK3R3
encoding the regulatory subunit 3 of PI3K.
ADTRP
-mediated up-regulation of
PIK3R3
activates AKT, resulting in up-regulation of
MIA3
. Knockdown of
ADTRP
expression promoted oxidized-LDL-mediated monocyte adhesion to endothelial cell (EC) and transendothelial migration of monocytes, inhibited EC proliferation and migration, and increased apoptosis, which was reversed by expression of constitutively active AKT1, while the over-expression of
ADTRP
in ECs blunted these processes. Knockdown of
MIA3
expression also promoted monocyte adhesion to ECs and transendothelial migration of monocytes. However, knockdown of
MIA3
increased
ADTRP
expression, suggesting that
MIA3
negatively regulates
ADTRP
expression. Genetically, no significant interaction between
ADTRP
SNP rs6903956 and
MIA3
SNP rs17465637 was detected in 2,185 CAD patients and 2,156 non-CAD controls by classical two-locus genotypic analysis, relative excess risk due to interaction or INTERSNP programs. We have recently found that one molecular mechanism for gene-gene interaction is positive cyclic cross-regulation of gene expression: Gene A positively regulates gene B, whereas gene B also positively regulates gene A. Here we have identified a different scenario in which gene A positively regulates gene B, but gene B negatively regulates gene A, thereby resulting in lack of gene-gene interaction. In conclusion, we have uncovered a novel molecular signaling pathway involving
ADTRP
and
MIA3
for the pathogenesis of CAD. We show that
ADTRP
positively regulates
PIK3R3
expression, which leads to activation of AKT and up-regulation of
MIA3
, thereby regulating endothelial cell functions directly relevant to atherosclerosis. These results further support that positive cyclic cross-regulation of gene expression is a molecular mechanism for gene-gene interaction.
Abstract 627: Human Osteopontin Isoforms Differentially Promote Neovascularization in Response to Ischemia via Macrophage Recruitment and Survival
Authors: Lee Grace S , Caroti Courtney M , Joseph Giji , Weiss Daiana , Salazar Hector F , Shin Yee Lok Zoe , Taylor W. Robert , Lyle Alicia N
Published: July 2021

Background:
Coronary and peripheral artery diseases result in vessel occlusion and ischemia, initiating neovascularization to restore blood flow and preserve function. We previously established that osteopontin (OPN), a matricellular cytokine, is critical to ischemia-induced neovascularization. Unlike rodents, humans express 3 OPN isoforms (a, b, and c); however, the roles of these isoforms in neovascularization and cell migration remain undefined.

Methods and Results:
Using a murine model of hindlimb ischemia in OPN
-/-
mice and 1.5x10
6
lentivirus particles expressing OPNa, OPNb or OPNc delivered IM, we found that OPN isoforms have different effects on functional perfusion recovery
in vivo
. OPNa increased limb perfusion 30.4%±0.8 and OPNc by 70.9%±6.3, as measured by laser Doppler perfusion imaging (d14; p<0.001 vs. LVGFP). Increases in perfusion translated to significant increases in functional limb use in OPNa and OPNc treated animals (61.1%±8.2; 76.2%±9.7; p<0.05), as assessed by voluntary running wheel use, and was not due to isoform expression differences (ELISA, n=6, p=ns). While OPN isoforms did not differentially affect angiogenesis, OPNa and OPNc significantly increased arteriogenesis (enlargement of arterioles), as measured by the increase in SM α-actin positive vessels in the small (200 - 700 μm
2
; 47.2%±6.1; 55.9%±6.7) and large artery (1000 - 2500 μm
2
; 54.2%±6.1; 76.5%±10.9) ranges
in vivo
(n=9; p<0.001 vs. OPNb). We hypothesized that OPN isoform-dependent effects on arteriogenesis are due to differential effects on macrophage function. OPN isoforms did not differentially affect macrophage polarization and all 3 isoforms increased macrophage survival (64.9%±1.1 - 78.6%±1.9 vs. control; p<0.0001). However, OPNa and OPNc both increased macrophage migration, where OPNc was the more potent migratory stimulus (n=4, p<0.001 vs. no trx, OPNa, OPNb).

Conclusion:
In conclusion, human OPN isoforms exert divergent effects on neovascularization through differential effects on arteriogenesis and macrophage migration and survival. Altogether, these data support that human OPN isoforms may represent novel therapeutic targets to improve neovascualrization and preserve tissue function in obstructive artery disease patients.
Abstract 633: Protein Kinase D Regulates VEGFR-2 Transcriptional Activity in Endothelial Cells Through AP-2β
Authors: Hoeppner Luke H , Bhattacharya Resham , Wang Ying , Angom Ramcharan Singh , Wang Enfeng , Dutta Shamit , Doeppler Heike R , Scarisbrick Isobel A , Storz Peter , Mukhopadhyay Debabrata
Published: July 2021

Vascular endothelial growth factor A (VEGF) signals primarily through its cognate receptor VEGFR-2 to control vasculogenesis and angiogenesis. Dysregulation of these physiological processes contributes to the pathologies of heart disease, stroke, and cancer. Protein kinase D (PKD) plays a crucial role in the regulation of angiogenesis by modulating endothelial cell proliferation and migration. In human umbilical vein endothelial cells (HUVEC) and human blood outgrowth endothelial cells (BOEC), knockdown of PKD-1 or PKD-2 downregulates VEGFR-2 and significantly inhibits VEGF-induced endothelial cell proliferation and migration. We sought to determine the molecular mechanism through which PKD modulates VEGFR-2 expression. Based on bioinformatics data, activating enhancer binding protein 2 (AP2) binding sites exist within the VEGFR-2 promoter. Thus, we hypothesized PKD may downregulate VEGFR-2 through AP2-mediated transcriptional repression of the VEGFR-2 promoter. Indeed, AP2β binds the VEGFR-2 promoter upon PKD knockdown in HUVEC as evident by chromatin immunoprecipitation assay. Luciferase reporter assays using serial deletions of AP2β binding sites within the VEGFR-2 promoter revealed transcriptional activity negatively correlated with the number of AP2β binding sites, thus confirming negative regulation of VEGFR-2 transcription by AP2β. Next, using siRNA, we demonstrated that upregulation of AP2β decreased VEGFR-2 expression and loss of AP2β enhanced VEGFR-2 expression.
In vivo
studies confirmed this finding as we observed increased VEGFR-2 immunostaining in the dorsal horn of the spinal cord of embryonic day 13 AP2β knockout mice. We hypothesize that PKD directly regulates AP2β function by serine phosphorylation and ongoing studies are being conducted to determine phosphorylation sites in AP2β directly regulated by PKD. Taken together, we demonstrate AP2β negatively regulates VEGFR-2 transcription and VEGFR-2 is a major downstream target of PKD. Our findings describing how PKD regulates angiogenesis may contribute to the development of therapies to improve the clinical outcome of patients afflicted by heart disease, stroke, and cancer.
Abstract 98: Chronic Kidney Disease-Associated Atherosclerosis is Attenuated by Dual Inhibition of microRNA-92a and microRNA-489
Authors: Wiese Carrie , Xu Zhi-Qi , Zhang Youmin , Guo Yan , Zhong Jianyong , Ramirez Marisol , Zhu Wanying , Kon Valentina , Vickers Kasey
Published: July 2021

Chronic kidney disease (CKD) subjects have an increased risk of developing cardiovascular disease, namely atherosclerosis. Endothelial dysfunction and inflammation are linked to the development of these diseases and recent work has identified a number of microRNAs (miRNAs) involved in these pathologies. As such, endothelial miRNAs are potential novel therapeutic targets to prevent and treat atherosclerosis. This study identified elevated aortic endothelial miR-92a-3p and miR-489-3p levels in a mouse model of CKD-associated atherosclerosis,
Apoe
-/-
mice with 5/6 nephrectomy. A combinatorial miRNA inhibition strategy resulted in the loss of both miR-92a-3p and miR-489-3p in the endothelium and significantly reduced the atherosclerotic lesion area by 33%. Total RNA sequencing of the aortic endothelium identified many altered genetic pathways and metabolic processes in response to
in vivo
miRNA loss-of-function, including inflammation, phospholipid metabolism, and protein degradation pathways. Results suggest that the reduction in atherosclerosis levels were not likely to be linked to alterations in plasma cholesterol levels or kidney function since these physiological parameters were not improved upon miRNA inhibition. Nevertheless, novel miRNA targets were identified to be significantly elevated in the aortic endothelium that may reduce inflammation leading to the improved physiological phenotype.
Fam220a
, a negative regulator of signal transducer and activator of transcription 3 (STAT3) phosphorylation, mRNA levels were significantly reduced in CKD-atherogenic mice compared to controls, but miRNA inhibition
in vivo
blocked the
Fam220a
repression. Moreover, gene reporter (luciferase) assays with site-directed mutagenesis confirmed
FAM220A
as a direct target of miR-92a-3p. Furthermore,
FAM220A
mRNA levels were repressed in human coronary artery endothelial cells (HCAEC) with miR-92a-3p over-expression, which resulted in increased phosphorylation of STAT3. Collectively, these results suggest that endothelial miR-92a-3p and miR-489-3p are novel therapeutic targets to treat CKD-associated atherosclerosis.
Abstract 12561: Validation of Score System Excluding Female Gender From CHA
2
DS
2
-VASc in Japanese Patients With Non-Valvular Atrial Fibrillation: A Subanalysis of the J-RHYTHM Registry
Authors: Tomita Hirofumi , Okumura Ken , Inoue Hiroshi , Atarashi Hirotsugu , Yamashita Takeshi , Origasa Hideki , Tsushima Eiki ,   J-RHYTHM Registry Investigators
Published: July 2021

Recent American and European guidelines recommend CHA
2
DS
2
-VASc (VASc) score for thromboembolic risk stratification in patients with non-valvular atrial fibrillation (NVAF), which includes female gender (Sc) as a risk. Although we validated this score in Japanese NVAF patients (Circ J 2014), our recent analysis showed that female gender is not a risk in this population (Am J Cardiol 2014). In the present analysis, we evaluated the score system excluding female gender from VASc in Japanese NVAF patients enrolled in J-RHYTHM Registry.

We prospectively assessed the incidence of systemic thromboembolism (SE) for 2 years in 997 Japanese NVAF patients without warfarin treatment (mean age, 68±12 years, 294 females). VASc score was determined, in which V was replaced by coronary artery disease (Co). Another score excluding female gender was defined as CHA
2
DS
2
-VA, or CHA
2
DS
2
-CoA (CoA). The predictive value of VASc and CoA scores for SE was compared by the c-statistics and the Net Reclassification Improvement (NRI). SE occurred in 7/294 females (1.2%/year) and 23/703 males (1.6%/year) (odds ratio 0.72, 95% confidence interval (CI) 0.28-1.62, p=0.44). Overall, there were no differences in the c-statistic and NRI between VASc and CoA scores (c-statistic 0.66, 95%CI 0.55-0.75, and 0.64, 0.54-0.74, respectively, p=0.76, and NRI 0.064, 95%CI -0.091-0.218, p=0.42). In male patients of VASc score 0, 1, and >2, SE occurred in 2/141 (0.7%/year), 4/194 (1.0%/year), and 17/368 (2.3%/year), respectively, and in females of VASc score 1 and >2, it occurred in 0/39 (0%/year) and 7/255 (1.4%/year), respectively. In female patients of CoA score 0, 1, and >2, SE occurred in 0/39 (0%/year), 1/51 (1.0%/year), and 6/204 (1.5%/year), respectively. There was a trend to be a significant NRI between the two scores in this stratification (NRI 0.059, -0.008-0.126, p=0.08). Finally, only in patients of VASc score 0 and 1 (n=374), there were significant differences in the c-statistic and NRI between VASc and CoA scores (c-statistic 0.52, 95%CI 0.32-0.72, and 0.58, 0.37-0.76, respectively, p<0.0001, and NRI 0.106, 95%CI 0.075-0.137, p<0.0001).

These findings indicate that CoA, a score excluding female gender from VASc, is useful for thromboembolic risk stratification in Japanese NVAF patients.
Abstract 003: Growth Trajectories of Body Mass Index During Childhood and Adult Hypertension
Authors: Li Shengxu , Zhang Tao , Wheltong Paul K , Krousel-Wood Marie , Bazzano Lydia , He Jiang
Published: July 2021

Background:
Childhood body mass index (BMI) predicts adult hypertension. However, information is lacking regarding the relationship between BMI growth trajectories during childhood and adult hypertension risk. We aimed to test the hypothesis that BMI growth rates at different childhood ages have differential influences on adult hypertension risk, independent of BMI levels.

Methods:
The study included 1,772 Whites and 960 Blacks with repeated BMI and blood pressure measurements from childhood (4-19 years) to adulthood (20-51 years). A random-effects mixed model was used to construct BMI growth curves by race and sex. Model-estimated linear growth rates of BMI at different childhood ages were linked to adult hypertension (defined by measured blood pressure or antihypertensive medication) in multivariable logistic regression models.

Results:
Hypertensive adults had higher BMI across childhood than normotensive adults. Overall, linear slope and cubic parameters of BMI were higher in hypertensive adults than in normotensive adults in all four race-sex groups (p ≤ 0.023). Odds ratio of adult hypertension for each kg/m
2
/year increase in BMI linear slope was 1.22 (95% CI =1.11-1.34) at age 4, was non-significant between ages 5-9, and gradually increased thereafter and plateaued beginning at age 15 (odds ratio = 1.41 and 95% CI = 1.28-1.56) (Figure).

Conclusions:
Rapid increase in BMI during and after puberty is associated with elevated risk of hypertension in adult life, which has implications for early prevention.
Abstract 007: Comparative Effectiveness of Blood Pressure Lowering Drugs in Reducing Cardiovascular Events: Results From a Network Meta-Analysis
Authors: Wei Jingkai , Galaviz Karla I , Kowalski Alysse , Magee Matthew J , Haw J. Sonya , Narayan K.M. Venkat , Ali Mohammed K
Published: July 2021

Introduction:
Blood pressure lowering drugs help prevent cardiovascular events though less is known about the comparative effectiveness of different drug classes. We aimed to compare the effect of different blood pressure lowering drugs classes on cardiovascular risk.

Hypothesis:
We hypothesized that the cardiovascular preventive effects of blood pressure lowering drugs would vary among different types of drug classes.

Methods:
PubMed, EMBASE, and Cochrane Library databases were searched for articles published between January 1, 1990 and June 30, 2014. Study eligibility criteria included randomized controlled trials testing blood pressure-lowering drugs, including angiotensin-converting-enzyme inhibitors, angiotensin receptor blockers, calcium-channel blockers, beta-blockers, and thiazide diuretics that reported cardiovascular outcomes (cardiovascular death, myocardial infarction, stroke, coronary revascularization and composite cardiovascular events) with at least 6 months of follow-up. Data from studies that met inclusion criteria were independently extracted by two reviewers using a standardized extraction form. For each outcome, a Frequentist, random effects network meta-analysis model was used to compare risk reductions between drug classes. The relative ranking probability of each blood pressure lowering drug class being the most effective was estimated using rankogram curves, surface under the cumulative ranking curves, and mean ranks.

Results:
A total of 102 trials that included 415,047 participants (mean age 65.1 years, 60.0% male) met inclusion criteria. Compared to placebo, blood pressure lowering drugs reduced the risk of composite cardiovascular events by 11-18% in aggregate. Calcium channel blockers were the most effective in reducing composite cardiovascular events risk (RR=0.82, 95% CI: 0.77, 0.88), stroke (RR=0.68, 95% CI: 0.61, 0.75) and revascularization (RR=0.84, 95% CI: 0.77, 0.92); angiotensin-converting-enzyme inhibitors were the most effective in reducing cardiovascular death (RR=0.85, 95% CI: 0.80, 0.91) and myocardial infarction (RR=0.83, 95% CI: 0.77, 0.91). Effects of drugs were influenced by blood pressure, where each 10mmHg reduction in systolic and diastolic blood pressure, regardless of drug class, was associated with a 2.5% (95% CI: -3.8, -1.2) and 5.4% (95% CI: -8.0, -2.4) lower risk of composite cardiovascular events, respectively.

Conclusion:
Angiotensin-converting-enzyme inhibitors had an advantage over other blood pressure lowering drugs in preventing myocardial infarction and cardiovascular death, while calcium channel blockers were more effective in preventing overall cardiovascular events, stroke and revascularization. Future studies should compare the effectiveness of combination of drug classes in reducing cardiovascular events.
Abstract 012: Capture-Recapture Using Multiple Data Sources: Estimating the Prevalence of Congenital Heart Disease
Authors: Duca Lindsey M , Khanna Amber D , Rausch Christopher M , Kao David , Crume Tessa
Published: July 2021

Introduction:
Improvements in the treatment of congenital heart disease (CHD) have resulted in the majority of infants born with CHD surviving into adulthood; completely modifying the epidemiologic profile of patients with CHD. Although the prevalence of CHD at birth has been robustly estimated, the prevalence of CHD in adolescents and adults in the U.S. is uncertain due to a lack of systematically collected population-based data. The unique disjointed healthcare system in the U.S. makes population-based surveillance of conditions like CHD difficult.

Hypothesis:
Use of capture-recapture methodology in a state-wide CHD surveillance system will result in a higher estimated prevalence of CHD in adolescents and adults by adjusting for incomplete case ascertainment.

Methods:
Adolescents and adults age 11 to 64 years with a CHD lesion listed as a diagnostic code on an encounter occurring between January 1, 2011 to December 31, 2013 were captured by the Colorado CHD surveillance system. Five primary data sources, representing electronic medical records (EMR) from participating healthcare systems and claims data from the All Payer Claims database, were used for case ascertainment. These sources provide inpatient, outpatient and emergency care across the state of Colorado. Once CHD cases were identified in one of the above data sources, a probabilistic record linkage algorithm was used for de-duplication of cases within and across data sources. Crude prevalence estimates were generated and then capture-recapture methods were employed to estimate the number of adolescents and adults with CHD in Colorado that were not captured in the surveillance system. Data were analyzed using a log-linear model incorporating severity of CHD as a variable of potential heterogeneous catchability.

Results:
The five primary data sources identified 24,907 CHD cases that met our case definition corresponding to 19,849 unique individuals during our 3-year surveillance period. The observed overall crude prevalence rate of CHD in adolescents and adults was 5.19 per 1000 population (95% CI 5.07 - 5.31 per 1000 population). Using capture-recapture methodology, the estimated prevalence of CHD in adolescents and adults corrected for incomplete case ascertainment was 5.68 per 1000 population (95% CI 5.59 - 5.77 per 1000 population), so an estimated 3,641 CHD cases were not identified in the five primary case finding data sources.

Conclusion:
Our study provides novel insight into strategies for EMR-based surveillance at the population-level by demonstrating the utility of capture-recapture methodology to estimate, and then correct for, cases missed in standard surveillance techniques.
Abstract 014: Comparative Cost-Effectiveness of 10-Year Atherosclerotic Cardiovascular Disease Risk Equations Over 10 Years of Follow-up: The Multi-Ethnic Study of Atherosclerosis
Authors: Tajeu Gabriel S , Kohli-Lynch Ciaran , Zhang Yiyi , Muntner Paul , Shea Steven , Moran Andrew
Published: July 2021

Introduction:
Uncertainty remains regarding the most efficient and cost-effective 10-year atherosclerotic cardiovascular disease (ASCVD) risk prediction tool for identifying moderate to high-risk patients for primary prevention statin treatment.

Methods:
We utilized the CVD Policy Model, a computer microsimulation model of ASCVD incidence, prevalence, mortality, and costs, to compare cost-effectiveness of statin treatment at varying 10-year predicted ASCVD risk thresholds for Framingham CVD (FRS-CVD), Reynolds Risk Score (RRS), and Pooled Cohorts Risk Equations over a 10-year time horizon in the Multi-Ethnic Study of Atherosclerosis (MESA) cohort. Cost effectiveness was assessed at predicted 10-year risk ≥ 20.0%, 15.0%, 10.0%, 7.5%, 5.0%, and 2.5%. We restricted the simulation cohort to participants aged 50 to 74 years who were not taking statins at baseline (n = 2,871). Moderate intensity statin treatment effectiveness was parameterized in the model as a 29% low-density lipoprotein cholesterol reduction. Total cost comprised statins ($100/year), side effect costs, and ASCVD event costs. Disability from treatment side effects and ASCVD events were included.

Results:
Average FRS-CVD, RRS, and Pooled Cohorts 10-year predicted ASCVD risks were 18.8%, 11.3%, 12.2%, for men and 8.9%, 4.3%, 6.6%, for women, respectively. At the same predicted risk, FRS-CVD consistently selected the most patients for treatment, and RRS the fewest (
Figure
). Compared with no treatment, treating patients with RRS ≥ 20% was cost saving in men. Subsequent risk threshold strategies with incremental cost effectiveness <$75,000/quality-adjusted life-year (QALY) for men were: FRS-CVD ≥ 20% ($13,046), RRS ≥ 7.5% ($17,774), and RRS ≥ 5.0% ($19,891). For women, the non-dominated thresholds were: Pooled Cohorts ≥ 15% ($27,908) and Pooled Cohorts ≥ 7.5% ($72,377).

Conclusions:
At cost-effectiveness thresholds less than $75,000/QALY, RRS was the highest value tool for men while the Pooled Cohorts Risk Equations performed best for women.
Abstract 028: Infant Growth Trajectories and Lipid Levels in Adolescence: Evidence From a Chilean Infancy Cohort
Authors: Von Holle Ann , North Kari E , Gahagan Sheila , Blanco Estela , Graff Misa , Justice Anne , Lozoff Betsy , Burrows Raquel , Green Howard Annie , Voruganti Saroja
Published: July 2021

Introduction:
Growth in early infancy is hypothesized to affect chronic disease risk factors later in life. To date, most reports draw on European ancestry cohorts with few observations of early growth. To determine if previous findings generalize to diverse study populations and to accommodate more detailed growth estimates, we investigated the association between monthly infant growth from birth to 5 months and dyslipidemia in adolescents in a Hispanic/Latino cohort.

Methods:
We characterized infant growth in males (n=345) and females (n=308) from the Santiago Longitudinal Study (SLS) using three metrics: weight (kg), length (cm) and weight-for-length (g/cm). Nonlinear mixed effects (SITAR) and latent growth mixture models (LGMM) were two approaches to estimate infant growth characteristics. Growth functioned as an exposure and lipid levels at 17 years were the outcome, including HDL-C, LDL-C, and TG. We used a false discovery rate of 0.05 to report findings.

Results:
Height trajectories presented the strongest evidence for an effect on dyslipidemia in adolescence in both the SITAR and LGMM models. SITAR analyses demonstrated an inverse relationship between height velocity before six months of age and HDL-C levels in adolescence. LGMM models offered more nuanced findings. A two-class height trajectory model offered the best fit: one group (n~268) had shorter length at birth accompanied by lower velocity/higher acceleration; the other group (n~406) had higher length at birth and higher velocity/lower acceleration. The lower velocity/higher acceleration group had lower mean HDL-C (mg/dL) (35.4, se=0.9) than the higher velocity/lower acceleration group (43.8, se=0.7), χ2(1)=36.4, p-value=<0.001. Similarly, the best-fitting 3-class weight trajectory model indicated the lower velocity/medium acceleration group had a higher mean LDL-C (mg/dL) (97.7, se=1.6, n~441) than the highest velocity/lowest acceleration group (88.8, se=2.0, n~202), χ2(1)=15.2, p-value=<0.001. A similar pattern for LDL-C emerged for the weight-for-length trajectories, χ2(1)=8.6, p-value=0.003.

Summary:
This study provides evidence of associations between infant growth from 0 and 5 months and blood lipid profiles during adolescence. Based on two different analytic approaches, characteristics of infant length trajectories were associated with HDL-C at mean age 17 years. These findings align with the well-established relationship between height and CVD in adulthood. Furthermore, groups with higher acceleration in all three trajectory types were more likely to have adverse lipid outcomes. Future research can inform the role of infant body size change in the context of downstream effects and CVD risk.