bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 304: Varicose Vein Development in Mice Is Blocked by Ator- and Rosuvastatin
Authors: Eschrich Johannes , Meyer Ralph , Pfisterer Larissa , Hecker Markus , Korff Thomas 
Published: July 2021

Venous diseases such as varicose veins and chronic venous insufficiency constitute a substantial source of morbidity. Despite their high prevalence, neither molecular mechanisms initiating these diseases nor pharmaceutic therapies have been explored so far. We recently reported that chronically increased filling pressure or biomechanical venous wall stress is sufficient to cause development of varicose-like veins in mice through a mechanism preceded by activation of the transcription factor activator protein 1 (AP-1). Considering these findings, we hypothesized that pharmacologic inhibition of AP-1 activity may attenuate venous remodeling. To prove this therapeutic concept, human venous smooth muscle cells (SMCs) were subjected to biomechanical stretch (13%, 0.5Hz) and treated with statins - HMG-CoA reductase inhibitors that are supposed to interfere with the activity of AP-1. EMSA and immunofluorescence studies revealed that ator- and rosu- but not simvastatin blocked stretch-induced AP-1 activity. Furthermore, ator- and rosuvastatin inhibited expression of the AP-1 target monocyte chemotactic protein 1 (MCP-1) and proliferation of these cells. In mouse veins exposed to an increased level of intraluminal pressure, atorvastatin diminished the abundance of MCP-1 and MMP-2 by ~90 % and inhibited proliferation of endothelial cells and SMCs (n=5, p<0.05). In line with this observation, growth of varicose-like veins in mice was suppressed by almost ~80 % upon treatment with rosu- and atorvastatin but not simvastatin (n=5, p<0.05) which was accompanied by a decrease in cellular proliferation as well as MMP-2 abundance in remodeling blood vessels. Consequent analyses of human varicose vein samples from patients chronically treated with statins indicated a decrease in SMC proliferation, MCP-1 and MMP-2 abundance (50-60 %, n=10, p<0.05).
Collectively, our current findings imply that both ator- and rosuvastatin effectively inhibit the development of varicose veins presumably by interfering with the wall stress-mediated activity of AP-1 in VSMCs. For the first time, this study revealed a possible pharmaceutic treatment that may be suitable to prevent growth of varicose veins and to limit formation of recurrences after varicose vein surgery.
Abstract 311: Network Analysis of Pathways Associated with Genetic Regulation of Trimethylamine-N-Oxide
Authors: Bennett Brian , Smallwood Tangi L , Wright Garron , Bostrom Meredith 
Published: July 2021

Common complex diseases, such as atherosclerosis, diabetes and obesity, are the result of genetic, environmental and gene/environmental interactions. Genetic reference panels are powerful tools to interrogate how the environment interacts with the underlying genetic architecture to influence disease susceptibility. Recently 2 genetic reference panels have been created: the Diversity Outbred (DO) mouse population and Collaborative Cross (CC). The mice comprising the CC/DO are mosaics of C57BL6/J, A/J, NOD/ShiLtJ, NZO/HiLtJ, WSB/EiJ, CAST/EiJ, PWK/PhJ, and 129S1/SvImJ. Both the DO and CC populations have tremendous genetic diversity, containing approximately 45 million segregating single-nucleotide polymorphisms (SNPs).
Our primary goal was to characterize the genetic regulation of risk factors for atherosclerosis susceptibility in the 8 founder strains of the DO/CC reference panels. Using a systems genetic approach, we perform integrative analysis of clinical traits, targeted metabolomics and RNA expression. We quantitated genome wide mRNA levels using RNA sequencing followed by network analysis. The network was then related to clinical traits and a targeted panel of 5 metabolites (trimethylamine-N-oxide (TMAO), Betaine, Choline and Creatinine) to identify key groups of genes (called modules). We performed weighted gene network co-expression analysis (WGCNA) using 17,122 genes across 46 mice (N = 5-8 mice per strain). The network contains 47 independent co-expression modules and associated these modules with plasma lipoproteins, liver triglyceride levels and the novel plasma risk factors TMAO and Betaine. For example, the turquoise module was most strongly associated with TMAO levels (p < 2 x 10-8) and the black module was associated with plasma Betaine concentrations. Both modules contain several candidate genes and enrichment analysis using Database for Annotation, Visualization and Integrated Discovery (DAVID ) v6.7 Studies are ongoing to validate the key molecular drivers of the network with a particular interest in the transcriptional network regulating plasma TMAO and Betaine concentrations.
Abstract 425: Lycopene Modulates T Lymphocyte Function by Modulating Th1 Responses and Treg Lymphocyte Population
Authors: Mills Lynsey M , Wilson Heather , Thies Frank 
Published: July 2021

Increased lycopene intake might have cardiovascular benefits, potentially through anti-inflammatory mechanisms. We recently showed that lycopene can influence lymphocyte activity by modulating processes involved in early cellular activation. T lymphocytes comprise different subsets, T cytotoxic, T helper 1 (Th1), T helper 2 (Th2) and T regulatory cells (Treg). We aimed to determine whether lycopene could specifically modulate T-cell subsets function and activity.
Peripheral blood mononuclear cells from 11 healthy adults were cultured for 18hr to 60h in the presence of lycopene-enriched liposomes (0-1.18μg lycopene/ml) with or without mitogens. The secretion of cytokines representative of Th1,Th2 and Treg activities were measured by ELISA (IL-2, IL-1β, IL-10, IFN-γ and TGF-β) or cytometric bead array (IL-4, IL-10, IL17 and IFN-γ). The population profile of Tc (CD3+/CD8+), Th (CD3+/CD4+), Treg (CD4+/CD25+), and the Treg subsets nTreg (CD4+/CD25+/FoxP3+) and iTreg (CD4+/CD25+/IL-10+) was determined by flow cytometry.
After 18h incubation, IL-2 concentration in the medium was significantly reduced (-29%, p=0.001) in the presence of lycopene (1.18μg/mL). Similar effects were observed after 36h and 60h culture for IFN-γ (-23%, p=0.015), Il-10 (-30%, p=0.023), IL-17 (-30%, p=0.019) but not IL-4 or TGF-β. The proportion of Treg cell was also significantly increased by 36% (p=0.001) in the presence of lycopene (1.18μg/mL) compared with non-treated activated cells. Furthermore, the proportions of iTreg cells were significantly increased by after incubation with lycopene while the proportion of nTreg cells decreased (-20.5 %, p=0.049).
We conclude that increased lycopene intake may be beneficial against atherogenesis by modulating T lymphocyte function, particularly in relation toTh1 and Treg.
Abstract 50: A Phase I Clinical Trial Evaluating the Safety of With Allogeneic Adipose Tissue-derived Multilineage Progenitor Cells-transplantation Therapy in Homozygous Familial Hypercholesterolemia Patients
Authors: Koseki Masahiro , Yamashita Shizuya 
Published: July 2021

Familial hypercholesterolemia (FH) is an inherited disorder, mainly caused by defects in low-density lipoprotein (LDL) receptor gene. The patients are characterized by high LDL cholesterol levels in the blood and premature cardiovascular disease. Although most of heterozygous FH patients are usually treated with statin, ezetimibe and bile acid sequestrants, homozygous FH patients are resistant to drug therapy. Therefore, in Japan, many of homozygous FH patients are treated by LDL-apheresis. LDL-apheresis is a great procedure to remove LDL cholesterol from the blood and contribute to improve prognosis of homozygous FH patients. However, the effect of removing LDL cholesterol is temporary and still not enough. As a definitive therapy, liver transplantation therapy could be one of options to recover LDL receptor, but donor is limited in Japan.
Therefore, based on the increase of the evidence about the safety of mesenchymal stem cells and percutaneous transhepatic portal approach in islet transplantation, we have developed a cell transplantation therapy with allogeneic adipose tissue-derived multilineage progenitor cells (ADMPCs), as an alternative treatment instead of liver transplantation.
Our group has already proved that xenogenic transplantation of human ADMPCs into Watanabe heritable hyperlipidemic rabbits resulted in significant reductions in total cholesterol, and the reductions were observed within 4 weeks and maintained for 12 weeks. These results suggested that hADMPC transplantation could correct the metabolic defects and be a novel therapy for inherited liver diseases.
Here, we report a protocol for the first-in-human clinical trial, which has been approved by the institutional review board and Ministry of Health, Labour and Welfare, Japan.
Abstract 542: The Effects of Soluble Epoxide Hydrolase Inhibition in Atherogenesis and Atherosclerotic Progression
Authors: Li Dan , Zhang Xu , Zhu Yi 
Published: July 2021

Epoxyeicosatrienoic acids (EETs), metabolites of arachidonic acid (AA), are known to have anti-inflammatory effects and hydrolyzed by soluble epoxide hydrolase (sEH). We aimed to investigate the effect of sEH inhibition on atherosclerotic progression.
LDLR-/- mice with or without sEH inhibitor and LDLR and sEH double knockout (DK) mice were fed with western diet for 6 weeks to induce arteriosclerosis, and then half of the mice switched to chow diet supplemented with sEH inhibitor for another 6 weeks. To further assess the role of sEH in atherosgenesis, we transplanted LDLR-/- or DK bone marrow into LDLR-/- recipients and also fed with 6 weeks western diet. The progression of atherosclerosis was evaluated and AA metabolites were analyzed by LC-MS/MS.
The results revealed that sEH inhibition and gene knockout decreased western diet-induced high plasma LDL cholesterol level and the size of the atherosclerotic lesion in the aorta of the mice. Furthermore, mice with DK bone marrow transplantation developed less aortic lesion by less ly6chi monocyte infiltiation without diminished the cholesterol level in plasma. After diet switch, the cholesterol level dropped back to control level and the size of arotic lesion became smaller compared to the western diet group. When sEH inhibitor decreased sEH activity in the liver and increased EETs/DHETs ratio in the plasma, the plaque size seemed not being further reduced after diet switch. However, M2 type macrophage marker at protein and mRNA level was increased in the aortic lesions. In cultured macrophage, sEH inhibition and EETs supplements could increase the markers of M2 macrophage at mRNA level.
Thus, sEH inhibition and depletion in macrophage attenuated atherosclerosis when fed with western diet. Although the role of the sEH inhibitor on atherosclerosis progression and plaque stability need to be further investigated. EETs levels and sEH activity may play a pivotal role in atherogenesis and monocyte/macrophage in filtration and differentiation.
Abstract 566: Experimental Investigation of Blood Flow in the Femoral Bifurcated Artery
Authors: Lotfi Azadeh , Lawler Zachary , Jan Omar , Barber Tracie , Simmons Anne 
Published: July 2021

Stent implantation is one of the most widely used interventional treatments for arterial stenosis which occurs predominantly due to atherosclerosis. Although stent placement can ensure very good patency of the lumen, stent-induced hemodynamic disturbance, which can lead to further stenosis, still remains a common clinical complication.
This study investigates the degree of hemodynamic disturbance induced by stenting an idealized bifurcated popliteal artery, which branches into the anterior and posterior tibial arteries, and is known as a site prone to atherosclerosis. Both stent-free and stented bifurcated arteries were examined, and the local flow patterns analysed for the comparative disturbance through the use of Micro Particle Image Velocity (micro-PIV) system. A life-size model of the artery was reconstructed using dimensions obtained from a patient specific MRI scan. The experiments were conducted under steady flow conditions, and the flow rates across the bifurcation were visualized and measured using the micro-PIV system.
It was shown that hemodynamic disturbances induced by the blood flow over the stent can further disrupt the arterial wall downstream of the stent causing further downstream vascular damage in addition to the in-stent restenosis. This downstream vascular disruption may require additional treatment depending on the type and severity of the damage. The results also support the hypothesis that links certain flow dynamic behaviour with the development of early intimal thickening, as the near wall low fluid momentum regions are found at locations where thickening was localized in bifurcated arteries in clinical studies.
Abstract 607: Kininogen Regulates Thrombin Generation in a Mouse Model of Sickle Cell Disease
Authors: Sparkenbaugh Erica M , Chandarajoti Kasemsiri , Key Nigel S , Gruber Andras , Mackman Nigel , McCrae Keith , Pawlinski Rafal 
Published: July 2021

Sickle cell disease (SCD) is associated with activation of coagulation and vascular inflammation. We previously demonstrated that, in mouse models of SCD, tissue factor (TF) expressed on leukocytes activates coagulation and contributes to inflammation via microvascular thrombosis, whereas non-coagulant form of TF expressed by endothelial cells mediates factor Xa (FXa)-PAR2 signalling and contributes to inflammation via IL-6 expression. We also showed that both rivaroxaban and dabigatran, oral inhibitors of FXa and thrombin, attenuated the hypercoagulable state in sickle mice.
It has been proposed that anticoagulants that target the intrinsic coagulation pathway may be associated with a lower risk of bleeding compared to targeting the extrinsic or common coagulation pathways. Therefore, we tested if the intrinsic pathway contributes to the activation of coagulation in sickle cell mice. Bone marrow from sickle (SS) and non-sickle (AA) Townes mice was transplanted into wild-type (WT) (n=18 AA, 15 SS), FXII-/- (n=14 AA, 22 SS), or FXI-/- (n=21 AA, 23 SS) recipients (all on C57Bl/6 genetic background). In addition, bone marrow from AA or SS mice was transplanted into high molecular weight kininogen (HK)+/+ (n=17 AA, 16 SS) or HK-/- (n=10 AA, 19 SS) mice. All mice were used for experiments 5 months after BMT.
Thrombin generation, measured by plasma thrombin anti-thrombin (TAT) levels, was increased in WT mice injected with SS bone marrow (WT/BMSS) compared to WT/BMAA mice (4.4±0.7 vs 2.6±0.3 ng/mL, p<0.05, mean ± S.E.M), however neither FXII nor FXI deficiency affected this parameter. Consistent with these data, inhibition of FXIIa-dependent activation of FXI with 14E11 antibody also did not reduce plasma TAT levels in SS Townes mice. Interestingly, elevated plasma TAT levels observed in HK+/+/BMSS mice was significantly reduced in HK-/-/BMSS mice (4.8±0.5 versus 3.48±0.2 ng/mL, p<0.05). These data indicate that HK, but not FXII and FXI, contributes to thrombin generation in SCD at steady state of disease. In vitro, HK fragments induce TF expression on monocytes via activation of CD11b/18. We are now investigating if inhibition of HK fragments-induced TF expression on monocytes may attenuate the hypercoagulabIe state in SCD mice without impacting hemostasis.
Abstract 637: Hyperhomocysteinemia-mediated sCD40L induction and CD16
+
CD40
+
Monocyte Differentiation in Chronic Kidney Disease
Authors: Yang Jiyeon , Choi Eric T , Kunapuli Satya P , Yang Xiaofeng , Wang Hong 
Published: July 2021

Chronic kidney disease (CKD) with uremia is associated with high mortality of cardiovascular disease (CVD) and Hyperhomocysteinemia (HHcy) is highly prevalent in uremic CKD patients. Elevated inflammatory monocyte (MC) is a cellular hallmark of chronic inflammation and CVD. Here we investigated mechanism in uremia-associated HHcy on MC differentiation in CKD-associated CVD. Data base mining revealed that CD40 is induced in MC from CKD subjects and associated with CVD, inflammatory disease and MC activation. Blood samples were obtained from 28 vascular disease patients with or without CKD.
By flow cytometric analysis using CD14+ as a MC marker, we observed inflammatory CD16+ MC is increased in CVD, whereas CD40+ MC and CD16+CD40+ MC are increased in CKD-associated CVD. CD40+ MC expresses T cell activation markers CD86, HLA-DR, adhesion receptor CD62L, and chemokine receptor Ccr2. Plasma CD40L levels are increased in CVD, positively correlated with CD16+ MC. Interestingly, plasma Hcy levels are increased in CKD-associated CVD, positively correlated with cellular Hcy, plasma creatinine, CD16+CD40+ MC, and negatively correlated with S-adenosylmethionine/S-adenosylhomocysteine (SAM/SAH), an indicator of methylation. In addition, MC and T cell inflammatory cytokines TNFα, IL-6, and IFN[[Unable to Display Character: &#612;]] are induced in CKD-associated CVD subjects.
Next, we examined mechanism of CD16+CD40+ MC differentiation using cultured human peripheral blood mononuclear cells (hPBMC). CKD serum, Hcy, and CD40L induced CD16+CD40+ MC differentiation, which were prevented by folic acid and CD40L antibody. IFN[[Unable to Display Character: &#1003;]], TNFα, and IL-6 synergistically induced CD16+CD40+ MC differentiation, which was blocked by neutralizing antibodies to TNFα and IL-6.
Hcy inhibited DNA methyltransferase 1 activity in isolated human blood MC. Finally, by gene analysis and pyrosequencing, we identified that the core promoter of CD40 gene is located at sole CpG island and hypomethylated at p65 consensus element in WBC from CKD-associated CVD subjects with low SAM/SAH ratio.
In conclusion, we identified CD16+CD40+ MC as a novel inflammatory MC subset which is increased in uremic HHcy-associated CVD. CD16+CD40+ MC differentiation may be due to CD40 promoter DNA hypomethylation.
Abstract 638: Flow Modulates the Endothelial Expression of Neuronal Guidance Cues
Authors: Zhang Huayu , de Bruin Ruben G , Dane Martijn J , Sol Wendy M , van der Veer Eric P , van den Berg Bernard M , Rabelink Ton J , van Zonneveld Anton Jan , van Gils Janine M 
Published: July 2021

Disturbed hemodynamic conditions lead to impaired homeostasis of the vascular endothelium and, subsequently, influx and retention of monocytes in the arterial wall. Emerging data suggest that, aside from a role in the development and maintenance of the nervous system, neuronal guidance cues (NGCs) are also required for physiological and pathological immune responses. We recently reported that NGCs are differentially expressed in atherosclerosis-prone and -resistant aortic sites in LDLR-/-mice during early atherogenesis. Here, we detail our investigation of the role of shear stress on the expression levels of NGCs (ephrins, slits, netrins and semaphorins) in human umbilical vein endothelial cells (ECs) under atherosclerosis-prone (static) and -protective (laminar flow) conditions.
To gain insight into the consequences of laminar flow on the mRNA expression levels of NGCs in ECs, the cells were exposed to 0, 1 or 7 days of laminar flow (shear stress 10 dynes/cm2, IBIDI-flow system). The mRNA levels of the leukocyte attractant NGCs EphrinA1, B1 and B2 were all reduced by laminar flow, in particular for EphrinB2 (52%[[Unable to Display Character: &#61615;]], p=0.02). In contrast, the potentially anti-inflammatory Slit2 displayed a striking early response to flow, with a marked increase in mRNA expression after 1day (2.2-fold, p=0.02), but was not sustained at Day 7. Interestingly, the expression levels of netrin-4, semaphorin 3F and semaphorin 4B were upregulated in ECs exposed to laminar flow for 1 and 7 days (2.1-fold, p=0.05; 3.0-fold, p=0.04; 2.1-fold, p=0.05; respectively), suggesting these NGCs as potentially athero-protective anti-inflammatory leukocyte repellent. Moreover, overexpression of the shear-induced transcription factor Krüppel-like factor 2 (KLF2) in static cultured ECs induced the expression of both semaphorin3F mRNA and protein (3.8 fold and 3.9 fold, respectively), while netrin-4 mRNA and protein upregulation appeared to be independent of KLF2.
Taken together, our study provides novel insight into regulation of the endothelial NGC expression profile by flow, which could implicate NGCs as mediators of the EC function by facilitating repulsion or attraction of monocytes under varying hemodynamic conditions.
Abstract 668: The Role of IL-27 Receptor Signaling in the Development of Abdominal Aortic Aneurysm
Authors: Peshkova Iuliia , Makhov Petr , Hou Vivianly , Koltsova Ekaterina 
Published: July 2021

Abdominal aortic aneurysm (AAA) is a cardiovascular disease, which is characterized by aortic wall dilation with subsequent rupture and internal bleeding. Inflammatory reactions in the vessel wall are likely essential for AAA pathogenesis, just like they are important for the development of another vascular pathology- atherosclerosis, which can predispose to AAA formation in a context-dependent manner. While inflammatory cytokines were shown to promote atherosclerosis and AAA, little is known about contribution of anti-inflammatory cytokines with regard to their ability to control vascular inflammation. Interleukin (IL)-27 signaling is required to suppress atherosclerosis development but its function in AAA remains unknown.
We utilize Angiotensin II (AngII) model to evaluate the role of IL-27R signaling in pathogenesis of AAA. AngII containing pumps were surgically implanted into Il27ra-/- x Ldlr-/-; Il27ra -/- x Apoe-/- and Il27ra+/- littermate controls and AAA progression was analyzed 4 weeks later.
Surprisingly, we found attenuated AAA progression in Il27ra-/- mice compared to Il27ra+/- and wt counterparts. The latter developed large AAA with visual hemorrhage into the artery wall, while Il27ra-/- mice developed small AAA with fewer myeloid cells and T cells. Moreover, opposite to aortic arches, T cells in abdominal aortas of Il27ra-/- mice produced less inflammatory IL-17A, while IFNγ production was unchanged. Interestingly, we found enhanced production of “Th2-like” cytokines IL-4 and IL-13, by NK cells and Type 2 Innate lymphoid cells (ILC2) in Il27ra deficient mice, which correlated with the protection from AAA.
Overall, we conclude that immunoregulatory cytokine IL-27 can differentially control atherosclerosis and AAA development by regulating innate and adaptive immune cell recruitment and cytokine production in the aortic wall.