bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 536: Prolyl-hydroxylase Inhibition as a Strategy of Therapeutic Angiogenesis
Authors: HoWangYin Kiave-Yune , Loinard Céline , Zouggari Yasmine , Cochain Clément , Récalde Alice , Richart Adèle , Smadja David M , D’Audigier Clément , Duong Van Huyen Jean-Paul , Bruneval Patrick , Emmerich Joseph , Silvestre Jean-Sébastien 
Published: July 2021

Therapeutic neovascularization represents an alternative treatment modality for patients with advanced ischemic coronary disease or critical limb ischemia (CLI).
Upregulation of HIF-1α, through specific PHDs inhibition, can be thought of as a master switch that coordinates the expression of a wide repertoire of genes involved in regulating vascular growth and remodeling. The aim of this study was to unravel the efficiency of specific prolyl-hydroxylase domain protein (PHDs) silencing in both cell and non cell-based therapeutic strategies in CLI.
We first showed that PHD2 and PHD3 mRNA levels were up-regulated whereas that of HIF-1α were down-regulated in mononuclear circulating cells and in ischemic legs of diabetic and non diabetic patients with CLI. We then used short hairpin RNAs (shRNAs) to downregulate murine PHD1, PHD2 or PHD3 levels and analyzed their efficiency as a therapeutic strategy in diabetic mice with hindlimb ischemia. PHDs silencing increased post-ischemic neovascularization. This increase was associated with an up-regulation of two key pro-angiogenic factors, VEGF-A and endothelial NOS and with that of pro-arteriogenic factors including fractalkine, MCP-1, and SDF-1 leading to activation of postischemic immunoinflammatory response. In addition, concomitant silencing of PHD2 and HIF-1α abrogated the shPHD2-related effects, demonstrating that activation of HIF-1α signaling mediated the pro-angiogenic and pro-arteriogenic effects of shPHD2. Finally, we assessed the putative beneficial effects of PHDs silencing on human endothelial progenitor cells (hEPC) or bone-marrow derived mesenchymal stem cells (hMSC)-based therapies. Cells were transfected with siRNA directed against PHDs and injected in Nude mice with hind-limb ischemia. We showed that PHDs silencing enhanced human cells therapeutic effect in the ischemic leg of Nude mice compared to animals receiving untreated cells.
In conclusion, our study reveals, for the first time, that silencing of PHDs by the transient and local upregulation of endogenous HIF-1α improves therapeutic angiogenesis. This study also paves the way for future strategy based on administration of siRNAs directed against PHDs to enhance the efficiency of cell-based strategy.
Abstract 545: Redox-Sensitive Regulation of Bone Marrow Hematopoietic Progenitor Cells
Authors: Pan Shi , Wang Nadan , Sheu Shey-Shing 
Published: July 2021

Bone marrow failure is a group of hematopoietic stem cell disorders that affect one or more lineages of blood cells. Hematopoietic progenitor cells are highly sensitive to reactive oxygen species (ROS)-induced oxidative stress. ROS impair the self-renewal of hematopoietic progenitor cells in the bone marrow and cause bone marrow failure. Glutaredoxin (Grx) is an antioxidant enzyme important for many cellular processes such as resistance against oxidative stress, DNA synthesis, sulfur assimilation, apoptosis, and cellular differentiation. However, the role of Grx in hematopoietic progenitor cells remains unknown.
We study the role of Grx in hematopoietic progenitor cells using Grx KO mice as the model system. Bone marrow cells were isolated from 9-10 weeks old wild type and Grx KO mice and the ratio of KSL cells, total blood count and p38 activation was studied. Our results showed no significant difference on the ratio of KSL cells between wild type and Grx KO mice in basal condition. There is also no significant difference in the numbers of red blood cells, lymphocytes, white blood cells and platelet in basal condition between wild type mice and Grx KO. Interestingly, upon hydrogen peroxide treatment, there is higher level of p38 activation in Grx KO mice compared to wild type mice. Our results suggest that Grx is involved in redox-sensitive regulation of hematopoietic progenitor cells. Grx may play a regulatory role in the prevention of bone marrow failure.
Abstract 101: Reconstituted High-density Lipoprotein - More Than the Sum of Its Parts.
Authors: Pirianov Grisha , Shoreim Azza , De Souza Ayesha , Torsney Evelyn , Cockerill Gillian 
Published: July 2021

Rationale
Reconstituted high-density lipoprotein (rHDL, CSL-111) particles have been shown to reduce multiple organ failure following hypovolemic shock, perturb irritable bowel syndrome and, more recently, to inhibit aneurysm formation in animal models. To identify new endothelial targets of the protective effect we examined the differential changes that occur in the proteome of activated primary endothelial cells following treatment with rHDLs.

Methods
Primary human endothelial cells were isolated from human umbilical cords, grown to confluence and activated with TNFα (10ng/ml), following a 4 hr incubation half the cells received rHDLs (1mg/ml, ApoAI) and the remaining cells received carrier only (10% sucrose). Protein lysates were prepared following 20 hr incubation and differential protein expression compared by scanning DiGE-2dimentional PAGE. Protein identities were confirmed using LC/MS of tryptic digests of proteins recovered from preparatory gels. Western blotting confirmed differential expression and established whether the response required the whole particle or solely the protein component of rHDLs.

Results
Of the proteins identified by LC/MS which were differentially expressed in response to rHDLs, Perilipin 3 (PLPN3, 1.3 fold, p<0.003); Growth factor receptor-bound protein 2 (GRB2, 2.5 fold, p<0.003) and Vimentin (1.6 fold, p<0.007) were induced, whilst sinexin 6 (SNX6, 1.2 fold p< 0.007); Glutathione S Transferase-1 (GST-1, 1.6 fold, p<0.02) and Crystallin-zeta like (CrystZL, 1.2 fold, p<0.004) were reduced. We found that the entire lipoprotein particle was necessary for the induction of PLN3 and reduction of SNX6. Concentration of rHDL < 100μg/ml were able to modulate the maximal effect observed. However, both rHDL and lipid free ApoAI were able to induce cytokine-inhibited expression of GRB2 (rHDL>ApoAI).

Conclusion
The PC component of rHDL is necessary for an acute effect on proteins involved in endosomal trafficking. Further studies are needed to understand the role this could play in the protective effect of HDL therapy.
Abstract 118: The Effect of CETP Inhibition With Anacetrapib on the Metabolism of PCSK9
Authors: Millar John S , Reyes-Soffer Gissette , Ginsberg Henry N , Rafeek Hashmi , Lassman Michael , Wong Allen , Jumes Patricia , Wagner John A , Gutstein David E , Rader Daniel J 
Published: July 2021

Introduction
Inhibition of CETP activity with anacetrapib has been associated with a significant reduction in low density lipoprotein (LDL)-cholesterol (C) levels both as a monotherapy and in combination with a statin. We recently reported that this reduction in LDL-C is due to a significant increase in the fractional clearance rate (FCR) of LDL apoB. One potential mechanism by which anacetrapib could influence LDL apoB clearance is through effects on PCSK9 metabolism. We conducted kinetic studies on PCSK9 to determine if a change in the metabolism of PCSK9 is responsible for the reduction in LDL-C levels following treatment with anacetrapib.

Methods
Moderately hyperlipidemic volunteers (n=39) were enrolled in a fixed-sequence study: 75% were on atorvastatin (20mg/day) plus placebo and 25% were on double placebo for four weeks. Following this baseline period subjects added anacetrapib (100 mg/day) to their background treatment for 8 weeks. Studies using D3 leucine tracer were performed at the end of each treatment period to determine the kinetics PCSK9.

Results
Statin treated subjects had lower LDL-C levels (84 vs 118 mg/dL) and higher PCSK9 levels (234 vs. 208 ng/mL) at baseline than subjects on placebo. The difference in PCSK9 was due to a higher PCSK9 production rate (PR) (19.7 vs. 16.7 ug/kg/day) but similar PCSK9 FCR (1.87 vs. 1.78 pools/day). Adding anacetrapib to background treatment was associated with a significant reduction in LDL cholesterol (~ -35%) and apoB levels (~ 20%) in both panels. Anacetrapib had no significant effect on PCSK9 levels in either of the background treatment groups. Consistent with the lack of change in PCSK9 levels, there was no effect of anacetrapib on the PR (18.1 vs. 19.3 ug/kg/day) or FCR of PCSK9 (1.82 vs. 1.93 pools/day).

Conclusion
These results indicate that in our study population the increase in LDL FCR observed in response to anacetrapib treatment is not likely to be related to changes in the metabolism of PCSK9.
Abstract 134: Kidney and Blood Pressure Abnormalities, and Cardiac Enlargement Accompany Moderate Coronary Artery Atherosclerosis in Western Diet Fed SR-BI/LDLR Double Knockout Mice
Authors: Fuller Mark T , Trigatti Bernardo 
Published: July 2021

Mice lacking the HDL receptor, SR-BI, are susceptible to spontaneous or diet-inducible occlusive coronary artery (CA) atherosclerosis and myocardial infarction (MI) in the context of absence, or reduced expression of apoE, respectively. Our lab has generated mice lacking both SR-BI and the LDL receptor (dKO mice). When fed diets that are very high in cholesterol, dKO mice develop rapid CA atherosclerosis and large transmural MIs, and exhibit significantly reduced survival. However, when fed a western type diet (WTD) with only moderately high cholesterol (22% fat, 0.125% cholesterol), dKO mice do not appear to exhibit reduced survival, and develop only mild CA atherosclerosis with little to no cardiac fibrosis. However they have significantly larger hearts than LDLR single KO (sKO) controls fed the same diet for the same period of time. We therefore examined additional factors that may account for the cardiac enlargement observed in these mice such as blood pressure (BP) and kidney pathology. 10 week old dKO mice and sKO controls were fed a WTD for 10 weeks; BP was measured before feeding, and periodically throughout feeding. Mice were euthanized after 10 weeks and blood and tissues were collected for analysis.
DKO mice had reduced cholesterol associated with VLDL and LDL sized lipoproteins compared to sKO control mice after 10 weeks of WTD feeding. As expected, heart weights were significantly increased in dKO mice compared to sKO controls. Additionally, dKO mice had significantly lower body weights compared to sKO controls treated equivalently. Before the onset of feeding, there was no significant difference in BP between dKO and sKO mice. After 1 week of WTD feeding, dKO mice had significantly increased BP compared to both pre-diet levels and to sKO controls fed the same diet for the same timeframe. However, BP in dKO mice dropped significantly after 9 weeks of WTD compared to pre-diet measurements, while BP in sKO mice was constant throughout the study. Oil red O staining of kidney sections revealed striking lipid deposition and enlargement of glomeruli in kidneys from dKO mice, this was not observed in sKO control mice.
Abstract 17: The Association Between Erythrocyte n-3 Polyunsaturated Fatty Acids (n-3 PUFAs) Content and Inflammation in Male Patients With Peripheral Artery Disease (PAD)
Authors: Grenon Marlene , Owens Christopher D , Alley Hugh , Chong Karen , Vittinghoff Eric , Harris William , Hughes-Fulford Millie , Rapp Joseph , Conte Michael 
Published: July 2021

Objectives
Dietary intake of n-3 PUFAs has been associated with cardiovascular disease, though the relationship to PAD is unclear. PAD patients have an increased burden of systemic inflammation. In a cross-sectional PAD cohort study, we evaluated the relationship between n-3 PUFAs content of red blood cells (omega-3 index) and biomarkers of systemic inflammation.

Methods
This was a prospective cohort study of patients (n=83) presenting to vascular surgery clinic for evaluation of PAD. The omega-3 index was measured using gas chromatography and inflammatory markers (hsCRP, IL-6, TNF-α and ICAM-1) via ELISA kits. We used linear regression to evaluate the independent association between the omega-3 index and inflammation.

Results
70 patients had PAD while 13 were found to have a normal ankle-brachial index (ABI). Mean (±SD) age was 67 ± 7 years. Mean ABI was 0.85 ± 0.23. The omega-3 index decreased across defined AHA hsCRP categories (Figure 1, p=0.04). One percentage point decrease in the omega-3 index was associated with increases in CRP (14%, 95% CI 0, 25, p=0.04), IL6 (8%, 95% CI 1, 15, p=0.02) and possibly ICAM-1 (4%, 95% CI -1, 12, p=0.13), but not TNF-α. After adjusting for age, race, HDL, smoking status, ABI and the body-mass index, the omega-3 index remained significantly (negatively) associated with systemic inflammation as measured by hsCRP in a male population at risk or suffering from PAD (p=0.05).

Conclusions
In a contemporary cohort of patients with PAD, the omega-3 index was negatively associated with biomarkers of inflammation. Our findings suggest a rationale for future studies of dietary manipulation of omega-3 index to reduce inflammation in patients with PAD.
Abstract 174: miR30c Regulates Plasminogen Activator Inhibitor-1 In Platelet
Authors: Luo Mao , Wan Qin , Liu Fei , Li Rong , Xia Jiyi , Ren Meiping , Chen Ni , Strawn Tammy , Fay William P , Wu Jianbo 
Published: July 2021

Objective.
PAI-1 mRNA and protein have been detected in human platelets. Recently some miRNAs have been found in human platelets, which are involved in the regulation of genes and the protein synthesis. However, little is known about the physiological roles of individual miRNAs in platelets. In this study, we investigated whether miR30c can regulate platelet-derived plasminogen activator inhibitor-1(PAI-1).

Methods and Results.
Expression of miR-30c, PAI-1, miR-21 and its targeted gene TIMP1 were found in healthy human leukocyte-depleted platelets (LDPs) by real time PCR. In luciferase reporter gene assay, miR-30c targets the 3’ untranslated region (3’ UTR) of PAI-1 mRNA through a miR-30c binding site. Transfection of miR-30c mimic into MEG-01, a megakaryoblastic cell line, significantly reduced PAI-1 protein level compared with negative control. Inhibition of miR-30c by transfecting miR-30c inhibitor significantly increased PAI-1 protein level. Furthermore, miR-21 expression was significantly down-regulated after transfecting with miR-30c mimic in PAI-1-/- mice LDPs, conversely, the expression of its target gene TIMP1 was significantly up-regulated after transfecting with miR-30c mimic in PAI-1-/- mice LDPs.

Conclusion.
These results provide a novel regulatory mechanism of miR30c- regulated PAI-1 protein through its influence on the downstream miR21 and its target gene TIMP1 expression in platelet, suggesting that miR-30c might be a potential new strategy for anti-thrombosis.
Abstract 181: BET Bromodomain Inhibition Suppresses Endothelial Inflammation and Atherosclerosis
Authors: Brown Jonathan , Duan Qiong , Griffin Gabriel , Paranal Ronald , Bair Steven , Newton Gail , Lichtman Andrew , Kung Andrew , Luscinskas Francis , Yang Tianlun , Croce Kevin , Bradner James , Plutzky Jorge 
Published: July 2021

Introduction
The BET bromodomain-containing family of proteins (BRD2, BRD3, BRD4) are epigenetic readers that coactivate transcription. Recent evidence indicates that BETs promote carcinogenesis and inflammation in sepsis, while BET bromodomain inhibitors are promising anti-cancer therapies. However, the role of chromatin remodeling in atherosclerosis in general and through BETs in particular remains unknown.

Hypothesis
We hypothesized that BET bromodomain-containing proteins coactivate proinflammatory responses in the vasculature with functional effects that promote atherogenesis.

Methods and Results

BET bromodomain inhibition, achieved with the highly selective, small-molecule inhibitor JQ1 significantly reduced early atherosclerosis (12 weeks) in cholesterol-fed, LDL receptor-null mice. In pursuing mechanisms for this effect, we identified BET protein expression in mouse and human endothelial cells (ECs) as well as endothelium from human atherosclerotic plaque. Treating human umbilical vein endothelial cells (HUVECs) with either JQ1 or siRNA to BRD2 or BRD4 potently suppresses TNFα-induced expression of adhesion molecules (SELE, VCAM1) and chemokines (CCL2, CXCL8). In chromatin immunoprecipation studies, TNFα stimulation of ECs recruited BETs to adhesion molecule and chemokine promoters coincident with RNA polymerase II and cyclin T1 localization, without altering NF-κB recruitment. In functional studies, JQ1 suppressed 1) monocyte adhesion to TNFα-activated HUVECs, 2) leukocyte rolling on cremaster post-capillary venules (intravital microscopy); 3) leukocyte transmigration (parallel-plate flow chamber); and 4) monocyte recruitment in thioglycolate-induced peritonitis
in vivo
.

Conclusions
BET bromodomain-containing proteins are novel determinants of pro-inflammatory transcription in the endothelium. Targeting chromatin by BET bromodomain inhibition may be a therapeutic strategy to limit atherosclerosis and other disorders involving endothelial inflammation.
Abstract 182: Quantitative Proteomics Reveals Dynamic Changes of Plasma Proteome in APOE-Deficient Mice at Four Phases of Atherosclerosis
Authors: Jing Linhong , Jin Zhu-qiu , Xie Wei , Zhang Shuaipeng J 
Published: July 2021

Introduction
To understand the mechanism for atherosclerotic disease at molecular level, we analyzed the dynamic changes of plasma proteome in mice at four different disease stages vs. control based on hypothesis that a panel of plasma proteins might associate with the progression of atherosclerosis.

Methods
Male APOE knockout (-/-) mice were fed with high fat diet and euthanized at 8, 12, 18 or 24 weeks of age, representing the early, intermediate, moderate and later stages of disease. Age-matched male wild type mice fed with high fat diet served as controls. At the time of euthanasia, aorta and plasma were collected. A mass spectrometry-based proteomics approach was employed to simultaneously quantify proteins in eight pooled samples.

Results
Image analysis of lesions in aortas (stained with Sudan IV) from 40 APOE -/- mice (10 mice at each time point) demonstrated the development of four different phases of disease (Fig.). No lesion was observed in the control mice. We have identified 57 dysregulated proteins (e.g., fibrinogen beta and gamma chains, complement C3, C4B, C9, apolipoproteins, etc.) at four disease stages. These protein’s expression levels, biological functions (e.g. immune modulation and inflammation), and pathways (e.g., complement and cogulation cascades) provide a better understanding of the basic molecular processes in atherosclerosis.

Conclusions
1) Proteomics analysis of plasma proteome changes at different stages of APOE-deficient mice facilitated the mechanistic studies of atherosclerosis. 2) A panel of plasma proteins was identified as potential “signatures” of atherosclerosis which may help early diagnosis and therapeutic intervention.
Abstract 19: Prognostic Value of Plasma Choline and Betaine Depend on the Intestinal Microflora-generated Metabolite Trimethylamine N-oxide
Authors: Wang Zeneng , Tang W. H. Wilson , Fu Xiaoming , Britt Earl B , Wu Yuping , Koeth Robert A , Levison Bruce , Hazen Stanley L 
Published: July 2021

Background
Recent metabolomics and animal model studies show trimethylamine-N-oxide (TMAO), an intestinal microbiota-dependent metabolite formed from dietary phosphatidylcholine (PC), is linked with coronary artery disease (CAD) pathogenesis. Prognostic determinants of other PC metabolites recently associated with prevalent CAD risk, such as choline and betaine, have not been described.

Methods
We examined the relationship between fasting plasma choline, betaine, and TMAO and future risk of major adverse cardiac events (MACE=death, myocardial infraction, stroke) over 3-year follow-up in 3,916 subjects undergoing elective coronary angiography. All analytes were quantified by stable isotope dilution LC/MS/MS.

Results
In our study cohort (mean age 63±11 years, 64% male), median [IQR] TMAO, choline, and betaine levels were 3.7 [2.4-6.2] μM, 9.8 [7.9-12.2] μM, and 41.1 [32.5-52.1] μM, respectively. Modest but statistically significant correlations were noted between TMAO and choline (r=0.33, p<0.001) and between TMAO and betaine (r=0.09, p<0.001). Higher plasma TMAO, choline and betaine levels were associated with a 2.7-fold, 1.9-fold, and 1.4-fold increased risk of MACE, respectively (Quartiles 4 vs 1: p<0.01, each). Following adjustments for traditional risk factors and hsCRP, elevated TMAO (Quartiles 4 vs 1: Hazard ratio, 1.97 [95% CI 1.50-2.60], p<0.01), choline (HR 1.34 [95% CI 1.03-1.74], p<0.05) and betaine levels (HR 1.33 [95% CI 1.03-1.73], p<0.05) each predicted increased MACE risk, but only TMAO predicted MACE risk when all 3 metabolites were included in the model (HR, 1.73 [95% CI 1.3-2.31], p<0.01). Choline and betaine only predicted increased MACE risk in those with elevated TMAO levels.

Conclusion
Elevated plasma levels of TMAO, and to a lesser extent choline and betaine, are associated with incident MACE risk. However, high choline and betaine levels are only associated with higher risk of future MACE with concomitant increase in TMAO. Collectively, these findings further support the atherogenic associations between intestinal-microflora dependent metabolism of PC and atherosclerosis in humans.
Abstract 234: Cigarette Smoke Induction of Mmp9 in Aortic Vascular Smooth Muscle Cells is Mediated by the Jak/stat Pathway
Authors: Ghosh Abhijit , Pechota Angela , Coleman Dawn , Upchurch Gilbert R , Eliason Jonathan L 
Published: July 2021

Introduction
Cigarette smoke has a strong correlation with abdominal aortic aneurysm (AAA) formation and is characterized by increased levels of MMP9 in the aorta. We hypothesized that smoke-induced MMP9 regulation is mediated by the Jak/Stat pathway thereby contributing to AAA formation.

Methods
Aqueous extract of cigarette smoke (AEC) was applied to male rat aortic vascular smooth muscle cells (RASMC) for 24 hours (h) in serum free medium (SFM), using SFM alone for controls. Media were collected for MMP2 and MMP9 zymography and proteins extracted from cells for phospho-Stat3 (pStat3), total Stat3 (T-Stat3), phospho-Jak2 (pJak2) and total Jak2 (T-Jak2) western blots. RASMC were transfected by siRNAs for Jak2 and Stat3 before AEC treatment to evaluate induction of MMPs. Coimmunoprecipitation and immunofluorescence microscopy investigated complex formation and cellular distribution of Jak2 and Stat3 following AEC treatment.

Results
MMP9 was increased in the medium of the AEC treated RASMC (P=0.005) and pro-MMP2 slightly elevated (P=0.055) at 24h compared to controls. By western blot protein extracts from AEC treated RASMC showed up regulation of pStat3, pJak2 and T-Jak2 and stable T-Stat3 compared with controls. siRNA transfection of RASMC for Jak2, Stat3 and a combination of Jak2 and Stat3 reduced MMP9 (P<0.005) and pro-MMP2 (P<0.05) in medium of AEC treated RASMC compared with AEC-treated cells transfected with control siRNA. Coimmunoprecipitation of total proteins from RASMC with total Jak2 antibody revealed increased levels of pStat3 in the AEC treated cells compared to controls, while separation of nuclear and cytoplasmic components followed by coimmunoprecipitation revealed more abundant Jak2/T-Stat3 complexes in the nuclear fraction than the cytoplasm following AEC treatment. Immunofluorescence microscopy revealed increased presence of pJak2, T-Jak2, pStat3 and T-Stat3 in the cytoplasm and nucleus of the AEC treated cells compared to the control cells.

Conclusion
This study suggests that cigarette smoke may result in AAA formation through Jak/Stat-mediated MMP9 production. siRNA inhibition of the Jak/Stat pathway greatly reduced AEC-induced MMP9 by RASMC and suggests a potential therapeutic target for the treatment of AAA.