Markers Of Vascular Inflammation Research Articles

Pericarotid Fat as a Marker of Cerebrovascular Risk.

Vascular inflammation is widely recognized as an important factor in the atherosclerotic process, particularly in terms of plaque development and progression. Conventional tests, such as measuring circulating inflammatory biomarkers, lack the precision to identify specific areas of vascular inflammation. In this context, noninvasive imaging modalities can detect perivascular fat changes, serving as a marker of vascular inflammation. This review aims to provide a comprehensive overview of the key concepts related to perivascular carotid fat and its pathophysiology. Additionally, we examine the existing literature on the association of pericarotid fat with features of plaque vulnerability and cerebrovascular events. Finally, we scrutinize the advantages and limitations of the noninvasive assessment of pericarotid fat.

Comparative Prognostic Value of Coronary Calcium Score and Perivascular Fat Attenuation Index in Coronary Artery Disease.

Coronary artery disease (CAD) is the leading global cause of mortality, accounting for approximately 30% of all deaths. It is primarily characterized by the accumulation of atherosclerotic plaques within the coronary arteries, leading to reduced blood flow to the heart muscle. Early detection of atherosclerotic plaques is crucial to prevent major adverse cardiac events. Notably, recent studies have shown that 15% of myocardial infarctions occur in patients with non-obstructive CAD, underscoring the importance of comprehensive plaque assessment beyond merely identifying obstructive lesions. Cardiac Computed Tomography Angiography (CCTA) has emerged as a cost-effective and efficient technique for excluding obstructive CAD, particularly in patients with a low-to-intermediate clinical likelihood of the disease. Recent advancements in CCTA technology, such as improved resolution and reduced scan times, have mitigated many technical challenges, allowing for precise quantification and characterization of both calcified and non-calcified atherosclerotic plaques. This review focuses on two critical physiological aspects of atherosclerotic plaques: the burden of calcifications, assessed via the coronary artery calcium score (CACs), and perivascular fat attenuation index (pFAI), an emerging marker of vascular inflammation. The CACs, obtained through non-contrast CT scans, quantifies calcified plaque burden and is widely used to stratify cardiovascular risk, particularly in asymptomatic patients. Despite its prognostic value, the CACs does not provide information on non-calcified plaques or inflammatory status. In contrast, the pFAI, derived from CCTA, serves as an indirect marker of coronary inflammation and has shown potential in predicting adverse cardiac events. Combining both CACs and pFAI assessment could offer a comprehensive risk stratification approach, integrating the established calcification burden with novel inflammatory markers to enhance CAD prevention and management strategies.

No effect of surgery on kidney and cardiovascular risk factors in mild primary hyperparathyroidism: secondary analyses from a 10-year randomized controlled trial.

Renal function and the skeleton are classic target organs in primary hyperparathyroidism (PHPT), affected by the chronic course of the disease. Most patients diagnosed today exhibit mild PHPT, characterized by slight hypercalcemia and no or unspecific symptoms. Concerns have been raised that PHPT could promote deteriorating kidney function and increase cardiovascular risk directly. To examine the effect of parathyroidectomy (PTX) on mild PHPT on renal function and markers for bone turnover, cardiovascular disease (CVD), and vascular inflammation. Prospective randomized controlled trial. ClinicalTrials.gov: NCT00522028. Eight Scandinavian referral centers. From 1998 to 2005, 191 patients with mild PHPT were included in Sweden, Norway, and Denmark. Of these 150 were included in the present analyses. Seventy patients were randomized to PTX and 80 to observation without intervention (OBS). e-GFR was calculated based on creatinine and cystatin C. Markers of CVD and systemic inflammation: osteoprotegerin, vascular cell adhesion molecule 1, soluble CD40 ligand, interleukin-1 receptor antagonist, von Willebrand factor. Bone turnover markers: C-terminal telopeptide of type 1 collagen (CTX-1) and serum Procollagen type 1 N-terminal propeptide. No differences in the development of renal function or vascular and systemic inflammation were detected. CTX-1 was lower in PTX after 10 years. Secondary analyses of a randomized controlled trial. PTX does not appear to affect renal function or markers of CVD and vascular inflammation in mild PHPT in a ten-year perspective.

C-reactive protein (CRP) and lactate dehydrogenase (LDH) as functional outcome predictors in stroke patients

Stroke is a neurologic disorder with high mortality and disability. Its pathophysiology is associated with vascular inflammation. However, studies between vascular inflammatory markers and stroke outcomes are still limited. This study aimed to investigate the association between inflammatory markers and functional outcomes of stroke. This was a retrospective cohort study involving all stroke patients at the Dr. Sardjito General Hospital, Yogyakarta from October 2020 to August 2021 who meet the inclusion and exclusion criteria. Mann-Whitney was used for bivariate analysis, followed by multivariate analysis. A total of 269 subjects, with 213 infarcts (79.2%) and 56 hemorrhagic (20.8%) strokes. There were 83 subjects deceased (30.9%), with 66 infarct (31%) and 17 hemorrhagic (30.4%) strokes. High CRP levels had significant and independent associations with worse GCS, ADL, IADL, NIHSS, BI, SSGM, MRS, and higher mortality rates (p < 0.05). High LDH levels had a significant and independent association with worse GCS scores and higher mortality rates (p < 0.05). Sub-analysis showed high CRP and LDH had associations with high mortality rates in infarct (p < 0.001), but only CRP (p = 0.029) had associations with high mortality rates in hemorrhagic. There was no significant association between fibrinogen and procalcitonin with stroke outcomes (p > 0.05). Cox-regression analysis showed CRP>24.5 mg/dL and LDH >300U/L associated with hazard ratios of 3.2 (p < 0.001) and 1.65 (p = 0.026). In conclusion. high CRP and LDH levels are associated with mortality rates in stroke patients.

Therapeutic effects of tilorone on mammary carcinogenesis through downregulation of pro-inflammatory cytokines and oxidative stress.

Tilorone dihydrochloride (tilorone) is an orally active interferon inducer with anticancer effects. The present study aimed to evaluate the anticancer effects of tilorone in breast cancer. MTT assay was done to measure the proliferation of MCF-7 and MDA-MB-231 breast cancer cells after treatment with tilorone. Mammary carcinogenesis was induced by subcutaneous injection (35 mg/kg, 0.5 mL) of dimethylbenz[a]anthracene (DMBA) in mammary pads of Sprague Dawley (SD) rats. Tumors were allowed to grow for 16 weeks till their sizes reached to 550-700 mm3, and then treated with 10 and 20 mg/kg of tilorone and standard drug doxorubicin (4 mg/kg) twice a week for 3 weeks. Normal and disease-control animals received normal saline. Tumor volumes and body weights were measured. Tumors were isolated to measure the levels of interferon-β (IFN-β), vascular endothelial growth factor-A (VEGF-A), P53 and inflammatory markers by enzyme-linked immunosorbent assay (ELISA). Serum biochemistry, lipid peroxidation (LPO) and antioxidant enzymes were measured by standard methods. Histopathology and immunohistochemistry (IHC) of P53 was done in tumor sections. Tilorone reduced the proliferation of MCF-7 and MDA-MB-231 cells with IC50 concentrations at 34.08 µM and 14.27 µM, respectively. Tilorone treatment showed reduced tumor volume, and increased survival with no significant changes in the body weights. Tilorone treatment also decreased levels of inflammatory markers and VEGF-A and increased IFN-β and P53 levels. Further, treatment with tilorone also decreased LPO and increased antioxidants levels. Histopathology of tumor sections showed normalizing morphology of treated animals. IHC of tumor sections showed increased levels of P53. In conclusion, tilorone has potential anticancer effects against breast cancer.

Pentraxin 3 as a marker of development and severity of stable coronary artery disease

PurposeInflammation plays a crucial role in the development of atherosclerotic plaques. Pentraxin 3 (PTX3) is produced at the site of inflammation and has been identified as a specific marker of atherosclerosis, vascular inflammation, and progression of the coronary artery disease (CAD).The aim of the study was to establish if PTX3 has potential relations with classical markers of cardiovascular risk, and if PTX3 may act as an independent risk factor of CAD occurrence and advancement. Materials and methodsThe study included 98 patients with stable CAD confirmed in coronary angiography (CAD group) (median age 65 interquartile range [IQR] 61–72 years; 72 ​% men). The control group consisted of 40 patients without CAD. ResultsThe CAD group had significantly higher PTX3 concentration compared to the control group. There was a correlation with age, male gender, lipid profile and intima-media thickness. There was no correlation between PTX3 concentration and the number of coronary vessels with significant atherosclerotic lesions and the advancement of atherosclerotic lesions on the Gensini scoring scale. The cut-off point was determined for 0.89 ​ng/ml for the exclusion of angiographically significant atherosclerotic lesions. ConclusionsPatients with CAD have significantly higher concentration of PTX3. There was no correlation between PTX3 and the advancement of angiographically significant atherosclerotic lesions in coronary arteries. Low PTX3 concentration may serve as an indicator for the absence of atherosclerosis.

Whole Body Vibration Training Has No Effect on Vascular Endothelial and Inflammatory Markers in Young Healthy Women.

Background: The aim of the study was to comparatively assess the impact of single and repeated whole body vibration training (WBVT) and training without vibration on changes in the concentration of vascular endothelial growth factor (VEGF), endothelial nitric oxide synthase (eNOS), and high-sensitivity C-reactive protein (hsCRP) in healthy, young, non-training women. Methods: The study involved 46 women (age 20.48 ± 1.72 years), who were divided into three groups: the experimental group participating in WBVT (EVG, n = 17); the comparison group performing the same exercises but without the vibration factor (EXG, n = 12); and the control group, which did not participate in any training (CON, n = 17). The program included participation in 36 training sessions conducted over 12 weeks, with a frequency of 3 times per week. In the EVG and EXG groups, venous blood was collected before and after the first and last training sessions, while in the CON group, blood was collected twice at a 3-month interval. Results: No significant changes were observed in the concentrations of the studied markers either after a single or repeated training session in both experimental groups (p > 0.05). Conclusions: The proposed WBVT protocol appears to be a safe form of exercise that does not induce negative inflammatory reactions. The applied vibration stimulus combined with physical exercises did not initiate pro-angiogenic processes or stimulate eNOS activity in healthy women, suggesting that similar studies should be conducted in individuals with circulatory problems or chronic inflammatory diseases.

Effects of resveratrol on changes in trimethylamine-N-oxide and circulating cardiovascular factors following exercise training among older adults

PurposeTrimethylamine-N-oxide (TMAO) is a gut-derived metabolite associated with cardiovascular disease (CVD). In preclinical and observational studies, resveratrol and exercise training have been suggested as potential strategies to reduce the systemic levels of TMAO. However, evidence from experimental studies in humans remains unknown. This project examined the dose-dependent effects of a combined resveratrol intervention with exercise training on circulating TMAO and other related metabolite signatures in older adults with high CVD risk. MethodsForty-one older adults [mean (±SD) age of 72.1 (6.8) years] participated in a 12-week supervised center-based, multi-component exercise training intervention [2×/week; 80 min/session] and were randomized to one of two resveratrol dosages [Low: 500 vs. High:1000 mg/day] or a cellulose-based placebo. Serum/plasma were collected at baseline and post-intervention and evaluated for TMAO and associated analytes. ResultsAfter the 12-week intervention, TMAO concentration increased over time, regardless of treatment [mean (±SD) Placebo: 11262 (±3970); Low:13252 (±1193); High: 12661(±3359) AUC; p = 0.04]. Each resveratrol dose produced different changes in metabolite signatures. Low dose resveratrol upregulated metabolites associated with bile acids biosynthesis (i.e., glycochenodeoxycholic acid, glycoursodeoxycholic acid, and glycocholic acid). High dose resveratrol modulated metabolites enriched for glycolysis, and pyruvate, propanoate, β-alanine, and tryptophan metabolism. Different communities tightly correlated to TMAO and resveratrol metabolites were associated with the lipid and vascular inflammatory clinical markers [|r| > 0.4, p < 0.05]. ConclusionThese findings suggest a distinct dose-dependent adaptation response to resveratrol supplementation on circulating metabolite signatures but not on TMAO among high-risk CVD older adults when combined with an exercise training intervention.

The beneficial effects of monoglucosyl hesperidin and monoglucosyl rutin on vascular flexibility: a randomized, placebo-controlled, double-blind, parallel-group study

Background: There is mounting evidence that the intake of polyphenols can help prevent cardiovascular complications. Furthermore, multiple intakes of different polyphenol compounds are expected to have a synergistic effect. However, few studies have examined the preventive effect of simultaneous polyphenol intake on cardiovascular complications in healthy adults. Objective: The present study aimed to evaluate the efficacy of daily intake of 70 mg of monoglucosyl hesperidin (MH) alone, or in combination with 140 mg of monoglucosyl rutin (MR), in improving vascular flexibility. Methods: This 8-week study was conducted on 66 healthy male and female participants with a relatively high body mass index (BMI) and low vascular flexibility. The participants were randomly assigned to an MH group (MH alone), MHMR group (combination of MH and MR), or placebo group by a computer-generated list (each n = 22). Participants took two tablets per day of either MH, MHMR or placebo during the intervention. Outcomes included vascular function indices, capillary flow, and inflammatory markers. Assessment points were at 4 weeks (4w) and 8 weeks (8w) of the intervention. Results: The primary outcome was flow-mediated dilation. In an analysis including participants whose left brachial-ankle pulse wave velocity (baPWV) was ≥1232.5 cm/s at baseline in the per-protocol set, the MHMR group showed significant improvement compared with the placebo group in flow-mediated dilation, maximum post-avascularization artery diameter, baPWV (each and average of both legs), and E-selectin at 8w. Conclusion: These results indicated that MH, in combination with MR, may act on vascular endothelial cells to improve vascular flexibility in healthy adults with a relatively lowvascular flexibility. (UMIN000046054). Keywords: Monoglucosyl hesperidin; Monoglucosyl rutin; flow-mediated dilation; E-selectin; pulse wave velocity, body mass index, vascular flexibility

Markers of vascular inflammation in pregnant women with gestational diabetes

Markers of vascular inflammation in pregnant women with gestational diabetes

Porphyromonas gingivalis Infection is Associated with Increased Vascular Inflammation in Patients with and Without Coronary Artery Disease

Vascular inflammation is a critical measure of risk for major adverse cardiac events in patients with and without known coronary artery disease. It is recognized that patients with poor oral hygiene tend to have an increased prevalence of coronary artery disease and myocardial infarction. Porphyromonas gingivalis (PG) is the keystone pathogen leading to periodontal disease. Despite evidence suggesting a link between active PG infection and coronary artery disease, there is limited data investigating whether PG can contribute to increased markers of vascular inflammation in patients with coronary artery disease. In this study, we studied 112 consecutive patients who presented at a community preventive cardiology clinic who consented to be assessed for the presence of PG. Subjects were enrolled in a mouth along assessment with measures of vascular inflammation, which was part of their routine clinical care. The average age of patients was 70.4 ± 8.5 years with 48.6% of the patients positive for PG and 20.5% with a history of coronary artery disease (CAD). The distribution of PG positivity was 48.3% and 34.8% in patients without and with a history of CAD, respectively. Patients with CAD have significantly greater levels of high sensitivity C-reactive protein (hsCRP_ (1.3 ± 0.3 vs. 3.2 ± 1.1 mg/dL, p = 0.04) and MPO (245.1 ± 19.4 vs. 371.4 ± 39.1 pmol/L, p = 0.003). Patients without a history of CAD, only Myeloperoxidase (MPO) was significantly increased. Statin use was significantly greater in those patients with a history of CAD. Interestingly, statin use tended to be greater in PG+ patients (71% vs. 62%) regardless of CAD status. The data from this early study demonstrate a link between active PG infection and increase in markers of vascular inflammation, especially in patients with established CAD. The findings suggest PG as a marker of cardiovascular risk, especially in patients with residual inflammatory risk.

Short, frequent, light-intensity walking activity improves postprandial vascular-inflammatory biomarkers in people with type 1 diabetes: The SIT-LESS randomized controlled trial.

To examine the effect of interrupting prolonged sitting with short, frequent, light-intensity activity on postprandial cardiovascular markers in people with type 1 diabetes (T1D). In a randomized crossover trial, 32 adults with T1D (mean ± SD age 28 ± 5 years, glycated haemoglobin 67.9 ± 12.6 mmol/mol, 17 women) completed two 7-h laboratory visits separated by >7 days. Participants either remained seated for 7 h (SIT) or interrupted sitting with 3-min bouts of self-paced walking at 30-min intervals commencing 1 h after each meal (SIT-LESS). Physical activity, insulin regimen, experimental start times, and meal consumption were standardized during each arm. Plasma levels of interleukin (IL)-1β, tumour necrosis factor (TNF)-α, plasminogen activator inhibitor (PAI)-1 and fibrinogen were sampled at baseline, 3.5 and 7 h, and assessed for within- and between-group effects using a repeated measures ANOVA. The estimated glucose disposal rate was used to determine the insulin resistance status. Vascular-inflammatory parameters were comparable between SIT and SIT-LESS at baseline (p > .05). TNF-α, IL-1β, PAI-1 and fibrinogen increased over time under SIT, whereas these rises were attenuated under SIT-LESS (p < .001). Specifically, over the 7 h under SIT, postprandial increases were detected in TNF-α, IL-1β, PAI-1 and fibrinogen (+67%, +49%, +49% and +62%, respectively; p < .001 for all). Conversely, the SIT-LESS group showed no change in IL-1β (-9%; p > .50), whereas reductions were observed in TNF-α, PAI-1 and fibrinogen (-22%, -42% and -44%, respectively; p < .001 for all). The intervention showed enhanced effects in insulin-resistant individuals with T1D. Interrupting prolonged sitting with light-intensity activity ameliorates postprandial increases in vascular-inflammatory markers in T1D. The trial was prospectively registered (ISRCTN13641847).

Expression of hTERT and Lp-PLA(2) genes is increased in primary hypothyroidism.

ObjectiveSightly low serum free T4 levels are associated with longer life expectancy. However, hypothyroidism is associated with increased risk of inflammation and vascular complications. Therefore, relation of thyroid hormones (THs) with life expectancy seems to be complex. This study was carried out in an effort to understand the contribution of THs in inflammation and aging. Methodology15 cases of treatment naïve primary hypothyroidism and 15 age and sex matched euthyroid controls were recruited. mRNA expression of Telomerase (hTERT), leucocyte telomere length (LTL) and Lipoprotein associated phospholipase A2 (Lp-PLA2), a marker of vascular inflammation and risk predictor of cardiovascular events were measured by qPCR in whole blood. ResultExpression of hTERT was found to be 6.95 times higher in the cases as compared to the controls. mRNA expression of Lp-PLA(2) was also 3.3 times higher in cases. LTL in euthyroid was approximately 81% of the length in hypothyroid patients. ConclusionA higher expression of hTERT indicates that hypothyroidism confers better cell survival in peripheral leucocytes inspite of the presence of vascular inflammation. However causal relationship cannot be ascertained with these results.

Higher Noncalcified Plaque Volume Is Associated With Increased Plaque Vulnerability and Vascular Inflammation.

Recently, it was reported that noncalcified plaque (NCP) volume was an independent predictor for cardiac events. Pericoronary adipose tissue (PCAT) attenuation is a marker of vascular inflammation and has been associated with increased cardiac mortality. The aim of this study was to evaluate the relationships between NCP volume, plaque vulnerability, and PCAT attenuation. Patients who underwent preintervention coronary computed tomography angiography and optical coherence tomography were enrolled. Plaque volume was measured by computed tomography angiography, plaque vulnerability by optical coherence tomography, and the level of coronary inflammation by PCAT attenuation. The plaques were divided into 2 groups of high or low NCP volume based on the median NCP volume. Among 704 plaques in 454 patients, the group with high NCP volume had a higher prevalence of lipid-rich plaque (87.2% versus 75.9%; P<0.001), thin-cap fibroatheroma (38.1% versus 20.7%; P<0.001), macrophage (77.8% versus 63.4%; P<0.001), microvessel (58.2% versus 42.9%; P<0.001), and cholesterol crystal (42.0% versus 26.7%; P<0.001) than the group with low NCP plaque volume. The group with high NCP volume also had higher PCAT attenuation than the group with low NCP volume (-69.6±10.0 versus -73.5±10.6 Hounsfield unit; P<0.001). In multivariable analysis, NCP volume was significantly associated with thin-cap fibroatheroma and high PCAT attenuation. In the analysis of the combination of PCAT attenuation and NCP volume, the prevalence of thin-cap fibroatheroma was the highest in the high PCAT attenuation and high NCP volume group and the lowest in the low PCAT attenuation and low NCP volume group. Higher NCP volume was associated with higher plaque vulnerability and vascular inflammation. The combination of PCAT attenuation and NCP volume may help identify plaque vulnerability noninvasively. URL: https://www.clinicaltrials.gov; Unique identifier: NCT04523194.

The BET inhibitor apabetalone decreases neuroendothelial proinflammatory activation in vitro and in a mouse model of systemic inflammation.

Brain vascular inflammation is characterized by endothelial activation and immune cell recruitment to the blood vessel wall, potentially causing a breach in the blood - brain barrier, brain parenchyma inflammation, and a decline of cognitive function. The clinical-stage small molecule, apabetalone, reduces circulating vascular endothelial inflammation markers and improves cognitive scores in elderly patients by targeting epigenetic regulators of gene transcription, bromodomain and extraterminal proteins. However, the effect of apabetalone on cytokine-activated brain vascular endothelial cells (BMVECs) is unknown. Here, we show that apabetalone treatment of BMVECs reduces hallmarks of in vitro endothelial activation, including monocyte chemoattractant protein-1 (MCP-1) and RANTES chemokine secretion, cell surface expression of endothelial cell adhesion molecule VCAM-1, as well as endothelial capture of THP-1 monocytes in static and shear stress conditions. Apabetalone pretreatment of THP-1 downregulates cell surface expression of chemokine receptors CCR1, CCR2, and CCR5, and of the VCAM-1 cognate receptor, integrin α4. Consequently, apabetalone reduces THP-1 chemoattraction towards soluble CCR ligands MCP-1 and RANTES, and THP-1 adhesion to activated BMVECs. In a mouse model of brain inflammation, apabetalone counters lipopolysaccharide-induced transcription of endothelial and myeloid cell markers, consistent with decreased neuroendothelial inflammation. In conclusion, apabetalone decreases proinflammatory activation of brain endothelial cells and monocytes in vitro and in the mouse brain during systemic inflammation.

Molecular imaging of large vessel vasculitis

Large vessel vasculitis (LVV) affects mainly large arteries with giant cell arteritis (GCA) and Takayasu arteritis (TAK) being the two most frequent forms. Clinical symptoms can be non-specific, including headache, fatigue, weight loss, and change in vision. Untreated, LVV may also lead to serious complications such as blindness, aortic aneurysm and dissection. Therefore, rapid recognition of the disease leading to accurate diagnosis and appropriate treatment is essential. FDG-PET/CT imaging has emerged as a sensitive marker of active vascular inflammation and its use in the management of LVV is now integrated in guidelines. In this article, we will discuss the role of FDG-PET/CT for the diagnosis of LVV and monitoring of therapy, as well as review technical and interpretation parameters.

The vitamin-k-cycle-enzyme VKORC1L1, but not its isoenzyme VKORC1, reduces inflammation in coronary endothelial cells

Abstract Introduction Vascular inflammation is a crucial contributor to atherosclerosis, in which oxidative stress, endoplasmic reticulum (ER) stress and transition of endothelial to mesenchymal cells (EndMT) are of critical importance. Vitamin-K-antagonists (VKA) promote vascular dysfunction, while vitamin K intake was proposed to reduce incidence of coronary artery disease. Key enzyme for regeneration of vitamin K is the Vitamin K epoxide reductase complex subunit 1 (VKORC1), which also represents the pharmacological target of VKA. VKOR-like1 (VKORC1L1) is an isoenzyme of VKORC1, which resides at the ER-membrane, exerts antioxidative properties and is involved in vitamin K maintenance. Aim of this study was to investigate the role of VKORC1 and VKORC1L1 in inflammation of human coronary artery endothelial cells (HCAEC). Methods and Results In silico analyses of the proteome of human coronary atherosclerosis revealed increased expression of VKORC1L1, but not of VKORC1. In vitro induction of oxidative stress (H2O2) and ER-Stress (tunicamycin) in HCAEC promoted time- and dose-dependent enhanced expression of VKORC1L1, without altering VKORC1 expression (H2O2: 2.3-fold vs. 0.8-fold after 40 minutes, p=0.04; tunicamycin: 1 μg/ml: 1.43-fold vs. 0.8-fold, p&amp;lt;0.01). Moreover, inducing EndMT by treatment with a differentiation medium (containing TGF-ß2, IL-1ß) resulted in a reduction of VKORC1L1, but not of VKORC1 (Fig.1). To analyze the relevance of the VKOR-enzymes on endothelial cell function, VKORC1 and VKORC1L1 were downregulated by siRNA. VKORC1L1 downregulation increased formation of reactive oxygen species (ROS) (DCFDA signal: 0.98-fold vs. 1.31-fold, p = 0.03) and reduced proliferation (EdU signal: 0.56 ± 0.02 vs. control, p &amp;lt; 0.01), whereas viability of HCAEC remained unchanged. ELISA and qPCR experiments revealed enhanced expression of a variety of markers of vascular inflammation after VKORC1L1 (e.g., IL-6: 3.42-fold ± 0.53, NF-κB: 1.95-fold ± 0.13, VCAM-1: 1.59-fold ± 0.12, vs. control). Additionally, expression of ER-stress markers was increased (e.g., GRP78; 1.32-fold ± 0.04 vs. control, p&amp;lt;0.01) (Fig.1). In contrast, VKORC1 downregulation significantly promoted proliferation (EdU signal: 3.9 ± 1.7 vs. control, p&amp;lt;0.01) as well as viability (alamarBlue® absorbance: 1.07 ± 0.08 vs. control, p=0.04), and decreased ROS (0.88 ± 0.17 vs. control, p=0.03). Pro-inflammatory and ER-stress proteins were either unaltered or decreased after VKORC1 knockdown (e.g., GRP78: 0.89-fold, VCAM-1: 0.68-fold) (Fig.2). To further elucidate these findings, we will perform overexpression experiments and stable knockdown of the VKOR-enzymes. Conclusion VKORC1 and its isoenzyme VKORC1L1 exhibit divergent effects on endothelial cell inflammation. Further studies are warranted to shed light on the regulation of the enzymes to improve our understanding regarding vascular side effects of VKA treatment and beneficial effects of vitamin K supplementation.

The endoplasmic reticulum (ER) chaperone GRP78 is secreted during ER Stress and alleviates endothelial cell inflammation

Abstract Introduction Glucose-Regulated Protein 78kD (GRP78) is a chaperone and the main regulator of the ER-stress response which is triggered by a variety of conditions that disturb folding of proteins in the ER. Upon ER Stress, GRP78 activates the unfolded protein response (UPR), which aims to clear unfolded proteins and restore ER homeostasis. Recently, extracellular secretion of GRP78 was described. However, the pathophysiological relevance of secreted GRP78 in atherosclerosis and endothelial cell inflammation remains to be elucidated. Aim of this study is to investigate the role of GRP78 secretion in endothelial cells. Methods and Results First, we sought to investigate if vascular cells secrete GRP78 during ER Stress. Human coronary artery endothelial cells (HCAEC) were treated with the ER stress inductor tunicamycin for up to 48h. After ER Stress induction, Western Blot and ELISA experiments detected an increased intracellular GRP78 expression. Intriguingly, prolonged ER Stress also promoted extracellular secretion of GRP78. Unbiased proteomic analysis of the HCAEC secretome confirmed that after ER-Stress induction, GRP78 is one of the most highly upregulated extracellular proteins (2.43-fold). Co-incubation with Brefeldin A, an inhibitor of ER-Golgi protein transport, abolished extracellular secretion (Fig.1). Hence, ER-Stress-induced GRP78 secretion is an actively regulated process. Next, the effect of GRP78 containing conditioned medium (CM) on HCAEC was analyzed. For control, HCAEC were co-incubated with BFA, or GRP78-knockdown was performed by siRNA. Treatment with GRP78 containing CM decreased GRP78 mRNA expression in target cells (0.35-fold vs. control [+BFA], p&amp;lt;0.0001). Furthermore, it increased viability ( BFA: 93.0 % vs. 83%, p=0.002; GRP78-siRNA: 94% vs. 85%, p=0.01) and decreased the rate of apoptosis (e.g. BFA: 1.7-fold vs. 5.1, p=0.02). Moreover, expression of markers of vascular inflammation and ER Stress (e.g., NF-κB and CHOP) were decreased when compared to CM without GRP78 (Fig.2). Furthermore, increased H2O2-generation was measured (BFA: 1.4-fold, p&amp;gt;0.001, GRP78-siRNA:1.3-fold, p=0.004) (Fig. 2). To confirm that the beneficial effects of the CM are in fact caused by GRP78, we treated HCAEC with recombinant GRP78 and observed an increased cell viability in a dose-dependent manner (1000 ng/ml: p=0.02 vs. control). CRIPTO is a membrane-bound co-receptor for growth factors and was previously linked to GRP78-signalling. siRNA-Knockdown of CRIPTO reduces viability and leads to cell apoptosis, irrespective of GRP78-treatment. Thus, GRP78-mediated effects may be mediated by CRIPTO. Conclusion Endothelial ER Stress promotes GRP78 secretion. Presence of GRP78 in conditioned medium ameliorates subsequent ER Stress and endothelial inflammation, which play a critical role in atherogenesis. Further investigations are important to shed light on the mechanism of release and the effect on target cells.

Abstract 13664: Relationship Between Vascular Inflammation Assessed by 18FDG-PET/CT and Coronary Calcification in Patients With Lung Cancer

Background: As the prognosis of lung cancer (LK) has improved over the years, atherosclerotic cardiovascular disease may affect the prognosis of LK. Previous studies have shown that vascular inflammation assessed by 18FDG-PET/CT and coronary artery calcification (CAC) assessed by chest CT can predict cardiovascular events. Objective: This study aimed to evaluate the relationship between vascular inflammation on 18FDG-PET/CT and CAC in patients with LK. Methods: Ninety-nine patients with LK who underwent both chest CT and 18FDG-PET/CT were included in this study. Vascular inflammation was assessed by 18FDG-PET/CT. Target-to-background ratio (TBR) was measured in 8 major arteries and was used as a marker of vascular inflammation. Agatston score was measured using chest CT images and was used as a marker of CAC. Results: The mean age was 70 years old (61 male). The median TBR was 1.39 (IQR: 1.33-1.43). Using a median value of TBR, patients were divided into 2 groups, high TBR group (≧1.39, n=50) and the low TBR group (&lt;1.39, n=49). High TBR group had more male gender (72% vs. 51%, p=0.03), was significantly older (71 (67-74)vs. 62(57-70)years, p&lt;0.001), and had diabetes (p=0.03). In contrast, the history of hypertension (p=0.993), dyslipidemia (p=0.622) and smoking(p=0.113) were similar between the groups. Agatston score was significantly higher in the high TBR group than in the low TBR group (168±338 vs. 56±97, p=0.005). Conclusion: Patients with lung cancer patients with high vascular inflammation had significantly higher Agatston score, suggesting an association between vascular inflammation and advanced coronary atherosclerosis. The impact of vascular inflammation and/or coronary calcification on the prognosis of LK needs further investigation.

FRI107 Ketogenic Diet Prevents Development Of Atherosclerotic Plaques In Apolipoprotein E Knockout Mice

Abstract Disclosure: V. Marzolla: None. A. Armani: None. C. Mammi: None. S. Gorini: None. E. Camajani: None. M. Caprio: None. Higher aldosterone (aldo) levels are associated with increased risk of cardiovascular ischemic events and mortality. It has been demonstrated that aldo accelerates the development of atherosclerosis in apolipoprotein E knockout mice (ApoE KO). Ketogenic diet (KD) positively impacts several cardiovascular risk factors, yet its effect on atherosclerosis is elusive. We hypothesize that KD protects from development of atherosclerotic plaques in ApoE KO mice, a murine model of atherosclerosis. Eight-week-old male ApoE KO mice were fed a KD (90,5% Kcal from fat, 0.4% Kcal from carbohydrate, 9.1% Kcal from protein) or a moderate high fat die (HFD) (42% Kcal from fat, 42.7% Kcal from carbohydrate, 15.2% Kcal from protein) and treated with aldosterone (aldo, 6 μg/mouse per day) or vehicle through osmotic mini-pumps. Cholesterol content was comparable in KD and HFD. After 4 weeks of treatment, in all experimental groups, intraperitoneal glucose tolerance test was performed, and peripheral blood samples were collected. We analysed cryosections of embedded aortic root to quantify the atherosclerotic plaque size, lipid and collagen content. We assessed vascular inflammation in speciments of thoracic aorta by analysing pro-inflammatory (ICAM-1, VCAM-1, IL-6, TNF-α, MCP-1) and anti-inflammatory (Arg-1, RETNLA, CCL5) genes. In ApoE KO mice treated with aldo, KD determined a significant improvement in glucose tolerance, compared to mice fed a HFD, without any significant effect on body weight. Beta-hydroxybutyrate levels were always significantly higher in KD-mice than in AD-mice, confirming nutritional ketosis in KD-mice. Analysis of aortic root showed that aldo treatment determined a significant increase in atherosclerotic plaque size and lipid content in HFD-mice. Such effects were significantly reduced in KD-mice, suggesting a favorable impact of ketosis in preventing atherosclerosis development. Plaque fibrosis did not differ among treatment groups. Finally, we observed a significant reduction in vascular inflammatory markers in KD-mice, when compared to HFD-mice. In particular, KD determined a significant reduction in gene expression of pro-inflammatory markers (ICAM-1, VCAM-1, IL-6, TNF-α, MCP-1) with the concomitant up-regulation of anti-inflammatory markers (Arg-1, RETNLA, CCL5), compared to HFD. The study suggests KD as a potential non-pharmacological approach to prevent the development of atherosclerotic disease in subjects with high cardiovascular risk. Presentation: Friday, June 16, 2023