Platelet Endothelial Cell Adhesion Molecule-1 Levels Research Articles

Targeted proteomics in a population-based study identifies serum PECAM-1 and TRIM21 as inflammation markers for periodontitis

ObjectivesPeriodontitis (PD) can cause systematic inflammation and is associated with various metabolic processes in the body. However, robust serum markers for these relationships are still lacking. This study aims to identify novel circulating inflammation-related proteins associated with PD using targeted proteomics.Materials and methodsWe used population-based, cross-sectional data from 619 participants of the Polish Longitudinal University Study (Bialystok PLUS). Mean pocket probing depth (mPPD) and proportion of bleeding on probing (pBOP) served as exposure variables. Fifty-two inflammation-related proteins were measured using the Olink Target 96 Cardiovascular III and the Olink Target 96 Immune Response panels. Associations between periodontal measures and proteins were tested using covariate-adjusted linear regression models.ResultsAt a false discovery rate of < 0.05, we identified associations of mPPD and pBOP with platelet-endothelial cell adhesion molecule-1 (PECAM-1) and tripartite motif–containing protein 21 (TRIM21).ConclusionThis study revealed novel associations between PD and serum levels of PECAM-1 and TRIM21. Our results suggest that these proteins might be affected by molecular processes that take place in the inflamed periodontium.Clinical relevanceNovel associations of PECAM-1 and TRIM21 with PD indicate promising serum markers for understanding the disease’s pathophysiological processes and call for further biomedical investigations.

Ozanimod Differentially Preserves Human Cerebrovascular Endothelial Barrier Proteins and Attenuates MMP-9 Activity Following In Vitro Acute Ischemic Injury.

Endothelial integrity is critical in mitigating a vicious cascade of secondary injuries following acute ischemic stroke (AIS). Matrix metalloproteinase-9 (MMP-9), a contributor to endothelial integrity loss, is elevated during stroke and is associated with worsened stroke outcome. We investigated the FDA approved selective sphingosine-1-phosphate receptor 1 (S1PR1) ligand, ozanimod, on the regulation/activity of MMP-9 as well as endothelial barrier components (PECAM-1, claudin-5, and ZO-1) in human brain microvascular endothelial cells (HBMECs) following hypoxia plus glucose deprivation (HGD). We previously reported that S1PR1 activation improves HBMEC integrity; however, mechanisms underlying S1PR1 involvement in endothelial cell barrier integrity have not been clearly elucidated. We hypothesized that ozanimod would attenuate an HGD-induced increase in MMP-9 activity which would concomitantly attenuate the loss of integral barrier components. Male HBMECs were treated with ozanimod or vehicle and exposed to 3h of normoxia (21% O2) or HGD (1% O2). Immunoblotting, zymography, qRT-PCR, and immunocytochemical labeling techniques assessed processes related to MMP-9 and barrier markers. We observed that HGD acutely increased MMP-9 activity and reduced claudin-5 and PECAM-1 levels, and ozanimod attenuated these responses. In situ analysis via PROSPER, suggested that attenuation of MMP-9 activity may be a primary factor in maintaining these integral barrier proteins. We also observed that HGD increased intracellular mechanisms associated with augmented MMP-9 activation, however ozanimod had no effect on these select factors. Thus, we conclude that ozanimod has the potential to attenuate HGD mediated decreases in HBMEC integrity in part by decreasing MMP-9 activity as well as preserving barrier properties.

Investigation of cardiovascular risk parameters in adolescents with metabolic syndrome.

Metabolic syndrome leading to type 2 diabetes mellitus and cardiovascular diseases is a chronic multifactorial syndrome, associated with low-grade inflammation status. In our study, we aimed at assessing the serum levels of follistatin (FST), pregnancy-associated plasma protein-A (PAPP-A), and platelet/endothelial cell adhesion molecule-1 (PECAM-1) in adolescent patients with metabolic syndrome. This study was performed in 43 (19 males, 24 females) metabolic syndrome adolescents and 37 lean controls matched for age and sex. The serum levels of FST, PECAM-1, and PAPP-A were measured by using ELISA method. Serum FST and PAPP-A levels in metabolic syndrome were significantly higher than those of controls (p < 0.005 and p < 0.05). However, there was no difference in serum PECAM-1 levels between metabolic syndrome and control groups (p = 0.927). There was a significant positive correlation between serum FST and triglyceride (r = 0.252; p < 0.05), and PAPP-A and weight, (r = 0.252; p < 0.05) in metabolic syndrome groups. Follistatin was determined statistically significant in both univariate (p = 0,008) and multivariate (p = 0,011) logistic regression analysis. Our findings indicated a significant relationship between FST and PAPP-A levels and metabolic syndrome. These findings offer the possibility of using these markers in diagnosis of metabolic syndrome in adolescents as the prevention of the future complications.

Novel subgroups in acute respiratory failure based on the trajectories of three endotheliopathy biomarkers: A cohort study.

Baseline levels of endotheliopathy are associated with worse respiratory outcomes and mortality in undifferentiated acute respiratory failure (ARF), but knowledge is lacking on the development of endotheliopathy over time in ARF. We, therefore, aimed to evaluate the prognostic significance of trajectories of endotheliopathy during the first days of ARF. We performed a secondary, exploratory analysis of a single-center prospective cohort including 459 patients requiring mechanical ventilation. Based on Days 1-3 Syndecan-1, soluble Thrombomodulin (sTM), and Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1), we divided patients into subgroups using latent class mixed modeling and correlated subgroups with clinical outcomes using Cox regression. Based on Syndecan-1 and sTM, respectively, we identified two subgroups. Based on PECAM-1, we identified three subgroups. Subgroups based on Syndecan-1 and sTM were identifiable from the baseline levels, but subgroups based on PECAM-1 were not. Patients with persistently high levels of both sTM and PECAM-1 were liberated from mechanical ventilation more slowly (Group high vs. Group low, sTM: hazard ratio [HR]: 0.66, 95% confidence interval [CI]: 0.50-0.88, p = .01, PECAM-1: HR: 0.59, 95% CI: 0.37-0.93, p = .02) and had higher 30-day mortality (sTM: HR: 1.90, 95% CI: 1.20-3.01, p = .01, PECAM-1: HR: 4.25, 95% CI: 1.99-9.07, p < .01). In ARF requiring mechanical ventilation, patients in subgroups with persistently high levels of sTM and PECAM-1 had lower rates of liberation from mechanical ventilation and higher 30-day mortality. However, patients with persistently high levels of sTM were identifiable based on the baseline level, and only the trajectory of PECAM-1 added information to that of the baseline level.

Endotheliopathy is associated with slower liberation from mechanical ventilation: a cohort study

BackgroundEndotheliopathy is suggested as pivotal pathophysiology of sepsis and trauma-associated organ failure, but its role in acute respiratory failure is not yet determined. We investigated if endotheliopathy biomarkers at ICU admission are associated with illness severity and clinical outcomes in patients with acute respiratory failure requiring mechanical ventilation.MethodsWe conducted a prospective single-center cohort study including 459 mechanically ventilated adults at ICU admission. Plasma levels of three endotheliopathy biomarkers were measured at ICU admission: Syndecan-1, soluble Thrombomodulin (sTM), and Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1). The primary outcome was the rate of liberation from mechanical ventilation, which is presented together with the rate of the competing risk of death while still on mechanical ventilation. Secondary outcomes were PaO2/FiO2-ratios on admission and on last measurement in patients dying within five days, and 30-day all-cause mortality. The primary outcome and 30-day all-cause mortality were analyzed using Cox regression, controlled for gender, age, chronic obstructive pulmonary disease, septic shock, heart failure, PaO2/FiO2-ratio at admission, respiratory infection, acute kidney injury, and bilirubin. PaO2/FiO2-ratios were analyzed using linear regression, controlled for age, chronic obstructive pulmonary disease, respiratory infection, and shock.ResultsPatients with high sTM were liberated from mechanical ventilation at a lower rate (adjusted hazard ratio (HR) 0.71, for an increase from the 25th to the 75th percentile, 95% confidence interval (CI) 0.54–0.93, p = 0.01). Patients with high PECAM-1 were liberated from mechanical ventilation at a lower rate, but only during the first 5 days (adjusted HR 0.72, for an increase from the 25th to the 75th percentile, 95% CI 0.58–0.9, p < 0.01). High levels of Syndecan-1 and PECAM-1 were associated with a higher rate of death while still on mechanical ventilation. sTM and PECAM-1 were negatively associated with PaO2/FiO2-ratio at ICU admission and no biomarker was associated with last measured PaO2/FiO2-ratio. High levels of all biomarkers were associated with higher 30-day all-cause mortality.ConclusionIn acute respiratory failure, endotheliopathy biomarkers are associated with lower rates of liberation from mechanical ventilation, hypoxemia at ICU admission, and 30-day all-cause mortality.Graphic

The role of tumor necrosis factor-α and interferon-γ in the hyperglycemia-induced ubiquitination and loss of platelet endothelial cell adhesion molecule-1 in rat retinal endothelial cells.

This study aimed to investigate the role of the hyperglycemia-induced increase in tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in the ubiquitination and degradation of platelet endothelial cell adhesion molecule-1 (PECAM-1) in the diabetic retina. Type I diabetes was induced in rats by the injection of streptozotocin, with age-matched non-diabetic rats as controls. Primary rat retinal microvascular endothelial cells were grown in normal or high glucose media for 6days or in normal glucose media for 24h with addition of TNF-α and/or IFN-γ. PECAM-1, TNF-α, IFN-γ, and ubiquitin levels were assessed using Western blotting, immunofluorescence, and immunoprecipitation assays. Additionally, proteasome activity was assessed both in vivo and in vitro. Under hyperglycemic conditions, total ubiquitination levels in the retina and RRMECs, and PECAM-1 ubiquitination levels in RRMECs, were significantly increased. Additionally, TNF-α and IFN-γ levels were significantly increased under hyperglycemic conditions. PECAM-1levels in RRMECs treated with TNF-α and/or IFN-γ were significantly decreased. Moreover, there was a significant decrease in proteasome activity in the diabetic retina, hyperglycemic RRMECs, and RRMECs treated with TNF-α or IFN-γ. Tumor necrosis factor-α and IFN-γ may contribute to the hyperglycemia-induced loss of PECAM-1 in retinal endothelial cells, possibly by upregulating PECAM-1 ubiquitination.

Platelet endothelial cell adhesion molecule-1 is a gatekeeper of neutrophil transendothelial migration in ischemic stroke

RationaleAdhesion molecules are key elements in stroke-induced brain injury by regulating the migration of effector immune cells from the circulation to the lesion site. Platelet endothelial cell adhesion molecule-1 (PECAM-1) is an adhesion molecule highly expressed on endothelial cells and leukocytes, which controls the final steps of trans-endothelial migration. A functional role for PECAM-1 in post-ischemic brain injury has not yet been demonstrated. ObjectiveUsing genetic Pecam-1 depletion and PECAM-1 blockade using a neutralizing anti-PECAM-1 antibody, we evaluated the role of PECAM-1 mediated trans-endothelial immune cell migration for ischemic injury, delayed brain atrophy, and brain immune cell infiltrates. Trans-endothelial immune cell migration was furthermore evaluated in cultured human cerebral microvascular endothelial cells. Methods and resultsTransient middle cerebral artery occlusion (tMCAO) was induced in 10–12-week-old male Pecam-1−/− and Pecam-1+/+ wildtype mice. PECAM-1 levels increased in the ischemic brain tissue due to the infiltration of PECAM-1+ leukocytes. Using magnetic resonance imaging, we observed smaller infarct volume, less edema formation, and less brain atrophy in Pecam-1−/− compared with Pecam-1+/+ wildtype mice. The transmigration of leukocytes, specifical neutrophils, was selectively reduced by Pecam-1−/−, as shown by immune fluorescence and flow cytometry in vivo and transmigration assays in vitro. Importantly, inhibition with an anti-PECAM-1 antibody in wildtype mice decreased neutrophil brain influx and infarct. ConclusionPECAM-1 controls the trans-endothelial migration of neutrophils in a mouse model of ischemic stroke. Antibody blockade of PECAM-1 after stroke onset ameliorates stroke severity in mice, making PECAM-1 an interesting target to dampen post-stroke neuroinflammation, reduce ischemic brain injury, and enhance post-ischemic brain remodeling.

Serum CXCL13 and PECAM-1 can be used as diagnostic and prognostic markers in elderly patients with gastric cancer

To investigate the application value of serum CXC Chemokine-13 (CXCL-13) and platelet endothelial cell adhesion molecule-1 (PECAM-1) in elderly patients with gastric cancer (GC). Ninety-eight elderly GC patients admitted to the Affiliated Hexian Memorial Hospital of Southern Medical University were selected as a research group, and 60 healthy subjects of the same age and in relatively good health who underwent physical examination at the same period were selected as a control group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of CXCL13 and PECAM-1 in serum. The clinical diagnosis and prognostic value of serum CXCL13 and PECAM-1 in elderly GC patients were analyzed. The levels of CXCL13 and PECAM-1 in serum of the research group were significantly higher than those of the control group (P < 0.001). The AUC value of combined diagnosis of elderly GC patients by serum CXCL13 and PECAM-1 was 0.950, and that of combined evaluation of prognosis of patients was 0.849. Serum CXCL13 and PECAM-1 were significantly related to TNM staging, differentiation degree and tumor diameter in elderly GC patients (P < 0.05). High levels of CXCL13 and PECAM-1 were significantly associated with lower 5-year OS (P < 0.05). Elderly GC patients with higher TNM staging, longer tumor diameters, high levels of CXCL13 and PECAM-1 had an increased risk of poor prognosis. Serum CXCL13 and PECAM-1 can be used as effective indicators for diagnosis and prognosis of elderly patients with GC, and can predict the 5-year OS in patients.

PECAM-1 protects against DIC by dampening inflammatory responses via inhibiting macrophage pyroptosis and restoring vascular barrier integrity

Disseminated intravascular coagulation (DIC) is a frequent complication of sepsis that affects patient outcomes due to accompanying thrombo-inflammation and microvascular permeability changes. Platelet endothelial cell adhesion molecule-1 (PECAM-1), a cellular adhesion and signaling receptor that is expressed on both hematopoietic and endothelial cells, plays an important anti-inflammatory role in acute and chronic inflammatory disease models. Little is known, however, about role and mechanism of PECAM-1 in septic DIC. Here, we investigated whether PECAM-1 might play a protective role in hindering the development of septic DIC. Plasma levels of soluble PECAM-1 were markedly elevated in septic patients that developed DIC, with a correspondingly poorer outcome. PECAM-1 knockout exhibited more severe DIC and poorer outcome in the LPS induced- and cecal ligation and puncture-induced DIC model, which could be alleviated by tissue factor inhibitor. This phenomenon seemed to be equally linked to PECAM-1 expression by both endothelial and blood cells. Furthermore, PECAM-1 was found to exert its protective effect on developing septic DIC by the following 2 distinct mechanisms: the inhibition of macrophage pyroptosis and the acceleration of the restoration of the endothelial cell barrier. Taken together, these results implicate PECAM-1 as a potentially attractive target for the development of novel therapeutics to manage and treat septic DIC.

S-endoglin expression is induced in hyperoxia and contributes to altered pulmonary angiogenesis in bronchopulmonary dysplasia development

Altered pulmonary angiogenesis contributes to disrupted alveolarization, which is the main characteristic of bronchopulmonary dysplasia (BPD). Transforming growth factor β (TGFβ) plays an important role during lung vascular development, and recent studies have demonstrated that endoglin is engaged in the modulation of TGFβ downstream signalling. Although there are two different isoforms of endoglin, L- and S-endoglin, little is known about the effect of S-endoglin in developing lungs. We analysed the expression of both L- and S-endoglin in the lung vasculature and its contribution to TGFβ-activin-like kinase (ALK)-Smad signalling with respect to BPD development. Hyperoxia impaired pulmonary angiogenesis accompanied by alveolar simplification in neonatal mouse lungs. S-endoglin, phosphorylated Smad2/3 and connective tissue growth factor levels were significantly increased in hyperoxia-exposed mice, while L-endoglin, phosphor-Smad1/5 and platelet-endothelial cell adhesion molecule-1 levels were significantly decreased. Hyperoxia suppressed the tubular growth of human pulmonary microvascular endothelial cells (ECs), and the selective inhibition of ALK5 signalling restored tubular growth. These results indicate that hyperoxia alters the balance in two isoforms of endoglin towards increased S-endoglin and that S-endoglin attenuates TGFβ-ALK1-Smad1/5 signalling but stimulates TGFβ-ALK5-Smad2/3 signalling in pulmonary ECs, which may lead to impaired pulmonary angiogenesis in developing lungs.

The Role of the Inflammatory Mediators TNF‐α and IFN‐γ in the Hyperglycemia‐induced PECAM‐1 Ubiquitination and Degradation in Diabetic Retinopathy

ObjectiveThe aim of this study was to investigate the role of TNF‐α and IFN‐γ in PECAM‐1 ubiquitination and subsequent degradation under hyperglycemic conditions in the diabetic retina.IntroductionInflammation is a major contributor to the development of diabetic retinopathy (DR). Two of the key mediators of inflammation, tumor necrosis factor‐α (TNF‐α) and interferon‐γ (IFN‐γ), have been shown to contribute to the pathophysiological changes leading to the progression of DR. However, their role in the hyperglycemia‐induced loss of retinal PECAM‐1 is not known. Platelet endothelial cell adhesion molecule‐1 (PECAM‐1) is a cell surface protein that is heavily expressed on vascular endothelial cells, and has an important role in the maintenance of the endothelium and its functions. We have previously found a significant decrease in PECAM‐1 levels in the diabetic retina and cultured rat retinal microvascular endothelial cells (RRMECs) grown under high glucose conditions. Thus, an investigation into the possible mechanism/s of PECAM‐1 loss may help in understanding the molecular processes leading to DR, and provide a possible therapeutic target.Materials and MethodsFor our in vitro studies, primary RRMECs were grown in normal glucose (NG, 5 mM glucose) or high glucose (HG, 25 mM) media for six days, with TNF‐α and/or IFN‐γ (20 ng/ml and 30 ng/ml, respectively) added 24 hrs prior to harvesting cells for analysis. For our animal studies, age‐matched male Wistar rats were injected with either vehicle (control), or Streptozotocin (STZ) (diabetic, 30 mg/kg/day) for three consecutive days in a model of type I diabetes. Retinas and plasma were collected eight weeks post‐STZ injection for analysis. PECAM‐1, TNF‐α, IFN‐γ, and ubiquitin levels were assessed using western blotting, ELISA, Immunofluorescence, and immunoprecipitation assays.ResultsTotal ubiquitination levels were significantly increased in the diabetic retina and in RRMECs grown under HG conditions (35% and 40% increase, respectively, p&lt;0.05). Moreover, PECAM‐1 ubiquitination levels were significantly increased in RRMECs grown under hyperglycemic conditions (3‐fold, p&lt;0.05). TNF‐α levels were significantly increased in diabetic STZ plasma and RRMECs grown under HG conditions (31%, 74.4% increase, respectively, p&lt;0.05). Additionally, IFN‐γ levels were significantly increased in the diabetic retina (35% increase, p&lt;0.05). PECAM‐1 levels were significantly decreased in RRMECs when treated with TNF‐α and/or IFN‐γ (60–75% decrease, p&lt;0.05), with a significant increase in total ubiquitination levels (20–28% increase, p&lt;0.05).ConclusionThese results indicate a possible role for TNF‐α and IFN‐γ in the hyperglycemia‐induced loss of PECAM‐1 in retinal endothelial cells, possibly by upregulating PECAM‐1 ubiquitination and subsequent degradation.Support or Funding InformationThis work was supported by funding from the National Institutes of Health (NIH) EY025632This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The Role of Matrix Metalloproteinase 2 (MMP2) In Diabetes‐induced Loss of PECAM‐1 in The Retina: Direct and Indirect Mechanisms

ObjectiveMatrix metalloproteinases (MMPs) are calcium‐dependent, zinc‐containing endopeptidases that are responsible for degrading extracellular matrix proteins, and for cleavage of various substrates such as cell surface receptors, proteoglycans, chemokines and cytokines. The aim of this study was to investigate the molecular mechanisms by which MMP2 may lead to PECAM‐1 loss in the diabetic retina, either directly through cleavage, or indirectly by modulating tumor necrosis factor‐α (TNF‐α) levels and activity.BackgroundPlatelet endothelial cell adhesion molecule‐1 (PECAM‐1) is a cell surface protein that is heavily expressed on vascular endothelial cells, and has major roles in barrier function, endothelial cell‐endothelial cell adhesion, and cell signaling. We have previously found a significant decrease (50–90%, P&lt;0.05) in PECAM‐1 levels in the diabetic retina, and in cultured rat retinal microvascular endothelial cells (RRMECs) grown under high glucose conditions (30–40%, P&lt;0.05). The decrease in PECAM‐1 levels was accompanied by a significant increase in MMP2 levels and activity in the diabetic plasma (72% and 78.5% respectively, P&lt;0.05), and in RRMECs grown under high glucose conditions (12‐fold increase, P&lt;0.05).Materials and MethodsFor our in vitro studies, primary RRMECs were grown in normal glucose (NG, 5 mM glucose) or high glucose (HG, 25 mM) media for six days, with treatments added 6–24 hrs prior to harvesting cells for analysis. For our animal studies, age‐matched male Wistar rats were injected with either vehicle (control), or Streptozotocin (STZ) (diabetic, 30 mg/kg/day) for three consecutive days in a model of type I diabetes. GM6001, a broad spectrum MMP inhibitor, was started at week six post‐STZ injection for two weeks (4 mM, 25 μl/eye, in sterile eye drops), and retinas were collected eight weeks post‐STZ injection for analysis. PECAM‐1, MMP2, and TNF‐α levels and interactions were assessed using western blotting, zymography, immunofluorescence, immunoprecipitation, or ELISA assays. In silico analysis for possible protease cleavage sites was done using PROSPER, provided by Monash University, Australia.ResultsSeveral PECAM‐1 cleavage sites by MMP2 were identified using in silico analysis. Moreover, we confirmed PECAM‐1/MMP2 interactions using immunoprecipitation. PECAM‐1 levels were significantly decreased in RRMECs treated with MMP2 (81%, P&lt;0.05), with a significant increase in TNF‐α levels (118.2%, P&lt;0.05). Moreover, RRMECs treated with TNF‐α had a significant decrease in PECAM‐1 levels (34%, P&lt;0.05). Furthermore, TNF‐α levels were significantly increased in diabetic STZ plasma, and in RRMECs grown under HG conditions (31%, 74.4% respectively, P&lt;0.05). To investigate the protective effect of MMP2 inhibition on PECAM‐1 levels, we treated diabetic rats with GM6001 eye drops, and saw an increase of PECAM‐1 levels (77.2%, P&lt;0.05) when compared to untreated rats.ConclusionThese results indicate a possible role of MMP2 in the diabetes‐induced loss of PECAM‐1 in the retina, which can be via direct cleavage of PECAM‐1, or indirectly via upregulation of TNF‐α, with the two mechanisms evaluated in future studies.Support or Funding InformationNIH EY025632, and AHA Predoctoral Fellowship 16PRE30080003This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Evaluation of ALCAM, PECAM-1 and selectin levels in intracranial meningiomas

ObjectivesCell adhesion molecules play a major role in various pathological states. The aim of this study was to evaluate the tissue levels of selectins (E-, L-, and P-Selectins), activated leukocyte cell adhesion molecule (ALCAM) and platelet endothelial cell adhesion molecule-1 (PECAM-1) in intracranial meningiomas and compare with the levels in control tissues. Patients and methods20 consecutive patients who were operated on meningiomas (grade-I: 17 and grade-II: 3) and 15 cerebral tissues obtained during the autopsy procedures as a resource for the healthy controls were included in this study. ResultsAll three selectins’, ALCAM and PECAM-1 levels were found to be significantly higher in meningiomas when compared with the control tissues (p<0.001, p<0.001, p<0.001, p<0.05, p<0.001), respectively. ConclusionsAccording to our results, the adhesion molecules were found to be higher in meningiomas suggesting that they may be involved in the pathological process of this type of brain tumors. We conceive that developing alternate therapies such as immunotherapeutic approaches against brain tumors might be amendatory in the treatment. Since this is the first study performed in meningioma type brain tumors demonstrating and comparing the levels of various adhesion molecules with control tissues, further clinical and experimental studies are needed to support our current findings with higher number of patients.

TOP2A, GGH, and PECAM1 are associated with hematogenous, lymph node, and peritoneal recurrence in stage II/III gastric cancer patients enrolled in the ACTS-GC study.

BackgroundTo identify factors related to relapse sites, we carried out an exploratory biomarker analysis of data from the Adjuvant Chemotherapy Trial of TS-1 for Gastric Cancer study, which is a randomized, controlled trial comparing postoperative adjuvant S-1 therapy with surgery alone in 1,059 patients with stage II/III gastric cancer.Patients and MethodsSurgical specimens from 829 patients were retrospectively examined, and 63 genes involved in a variety of biological processes were analyzed by quantitative real-time PCR. Gene expression normalized to reference genes was categorized as lower or higher than the median, and association with relapse sites was analyzed based on 5-year relapse-free survival.ResultsHematogenous, lymph node, and peritoneal recurrence developed in 72, 105, and 138 of the 829 patients, respectively; hazard ratios were 0.79 (95% confidential interval: 0.54–1.16), 0.51 (0.31–0.82), and 0.60 (0.42–0.84), respectively. Expression of platelet/endothelial cell adhesion molecule 1 (PECAM1) and topoisomerase II alpha (TOP2A) was strongly correlated with hematogenous recurrence and peritoneal recurrence, respectively (false discovery rate = 7.7×10−5 and 0.002, respectively). Gamma-glutamyl hydrolase (GGH) expression was moderately correlated with lymph node recurrence (false discovery rate = 0.34). Relapse-free survival was worse in patients expressing high levels of PECAM1 (hazard ratio = 2.37, 1.65–3.41), TOP2A (hazard ratio = 2.35, 1.55–3.57), or GGH (hazard ratio = 1.87, 1.13–3.08), respectively. A multivariate analysis revealed that these were stronger independent risk factors than tumor histological type.ConclusionIn patients with stage II/III gastric cancer, TOP2A, GGH, and PECAM1 levels in primary tumors are linked to high risk of hematogenous, lymph node, and peritoneal recurrence, respectively.

Serum levels of platelet endothelial cell adhesion molecule-1 (PECAM-1) and soluble vascular endothelial growth factor receptor (sVEGFR)-1 and -2 in HIV associated preeclampsia

ABSTRACTObjective: The angiogenic–antiangiogenic imbalance evident in preeclampsia (PE) may be used as a predictive tool to identify women likely to develop the clinical features in early pregnancy. Method: This retrospective study examined normotensive pregnant (n = 38) and preeclamptic (n = 38) HIV-infected and uninfected women to quantify sVEGFR-1 and -2 and PECAM-1 levels. Results: In contrast to PECAM-1, sVEGFR-1 and -2 differed according to pregnancy type (p = 0.07; p = 0.001; p = 0.002) but not by HIV status (p = 0.68; p = 0.13; p = 0.43). Conclusion: Irrespective of the HIV status, we report an upregulation of sVEGFR-1 with concomitant decline of PECAM-1 and sVEGFR-2 levels in PE compared to normotensive pregnancies.

Synergistic Effect of TNF-α and Dengue Virus Infection on Adhesion Molecule Reorganization in Human Endothelial Cells.

Dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) is a severe pathological manifestation of dengue virus (DENV) infection. Enhanced production of cytokines in dengue patients is proposed to induce endothelial barrier instability resulting in increased vascular leakage. Tumor necrosis factor (TNF)-α is an inflammatory cytokine that activates endothelial cells and enhances vascular permeability and plasma leakage in DHF/DSS. The present study investigated the in vitro effect of TNF-α and DENV infection on the expression of adherence junction proteins, tight junction proteins, and membrane integrity of human endothelial cell lines. Immunofluorescence staining and western blot analysis demonstrated platelet endothelial cell adhesion molecule-1 (PECAM-1) reorganization and decreased levels of the tight junction protein occludin in human endothelial cells treated with TNF-α and DENV, compared to mock, DENV, or TNF-α-treated cells. Permeability assessed by FITC-dextran as a transport molecule was increased and correlated with the unusual reorganization of PECAM-1. The altered distribution of PECAM-1 and low occludin protein levels in human endothelial cells treated with TNF-α and DENV correlated with increased permeability. In conclusion, the synergistic effect of TNF-α and DENV induced permeability changes in endothelial cells. These results contribute to the understanding of the mechanisms underlying enhanced vascular permeability in DENV infection.

Delayed Microvascular Shear Adaptation in Pulmonary Arterial Hypertension. Role of Platelet Endothelial Cell Adhesion Molecule-1 Cleavage.

Altered pulmonary hemodynamics and fluid flow-induced high shear stress (HSS) are characteristic hallmarks in the pathogenesis of pulmonary arterial hypertension (PAH). However, the contribution of HSS to cellular and vascular alterations in PAH is unclear. We hypothesize that failing shear adaptation is an essential part of the endothelial dysfunction in all forms of PAH and tested whether microvascular endothelial cells (MVECs) or pulmonary arterial endothelial cells (PAECs) from lungs of patients with PAH adapt to HSS and if the shear defect partakes in vascular remodeling in vivo. PAH MVEC (n = 7) and PAH PAEC (n = 3) morphology, function, protein, and gene expressions were compared with control MVEC (n = 8) under static culture conditions and after 24, 72, and 120 hours of HSS. PAH MVEC showed a significantly delayed morphological shear adaptation (P = 0.03) and evidence of cell injury at sites of nonuniform shear profiles that are critical loci for vascular remodeling in PAH. In clear contrast, PAEC isolated from the same PAH lungs showed no impairments. PAH MVEC gene expression and transcriptional shear activation were not altered but showed significant decreased protein levels (P = 0.02) and disturbed interendothelial localization of the shear sensor platelet endothelial cell adhesion molecule-1 (PECAM-1). The decreased PECAM-1 levels were caused by caspase-mediated cytoplasmic cleavage but not increased cell apoptosis. Caspase blockade stabilized PECAM-1 levels, restored endothelial shear responsiveness in vitro, and attenuated occlusive vascular remodeling in chronically hypoxic Sugen5416-treated rats modeling severe PAH. Delayed shear adaptation, which promotes shear-induced endothelial injury, is a newly identified dysfunction specific to the microvascular endothelium in PAH. The shear response is normalized on stabilization of PECAM-1, which reverses intimal remodeling in vivo.

PECAM-1 gene polymorphism (rs668) and subclinical markers of carotid atherosclerosis in patients with type 2 diabetes mellitus.

ABSTRACTThe platelet endothelial cell adhesion molecule 1 (PECAM-1) plays an important role in many inflammatory processes, including the development of atherosclerosis. Polymorphism rs668 of the PECAM-1 gene (373C/G) is functional, and it was reported to be associated with increased serum levels of PECAM-1. We investigated the association between the rs668 polymorphism of PECAM-1 and subclinical markers of carotid atherosclerosis in subjects with type 2 diabetes mellitus (T2DM). Five hundred and ninety-five T2DM subjects and 200 control subjects were enrolled. The carotid intima-media thickness (CIMT) and plaque characteristics (presence and structure) were assessed ultrasonographically. Biochemical analyses were performed using standard biochemical methods. Geno-typing of the PECAM-1 gene polymorphism (rs668) was performed using KASPar assays. The control examinations were performed 3.8 ± 0.5 years after the initial examination. Higher CIMT was found in patients with T2DM in comparison with subjects without T2DM. Statistically sig-nificantly faster progression of the atherosclerotic markers was shown in subjects with T2DM in comparison with the control group. When adjusted to other risk factors, the rs668 GG genotype was associated with an increased risk of carotid plaques in subjects with T2DM. We concluded that our study demonstrated a minor effect of the rs668 PECAM-1 on markers of carotid atherosclerosis in subjects with T2DM.

Platelet endothelial cell adhesion molecule-1 gene 125C/G polymorphism is associated with deep vein thrombosis.

Deep vein thrombosis (DVT) is a common disorder that is associated with high morbidity and mortality. Genetic factors have been suggested to influence the predisposition towards thrombosis and the incidence of DVT. Platelet endothelial cell adhesion molecule-1 (PECAM-1) is a key adhesion molecule that is involved in platelet function and maintenance of endothelial cell junctions. To date, no studies have examined the association between polymorphisms in PECAM-1 and DVT. The present study analyzed the single nucleotide polymorphisms (SNPs) of PECAM-1, namely Leu125Val (C373G), Asn563Ser (T1688C) and Gly670Arg (C2008T), in Chinese patients with DVT and age-and gender-matched controls, using polymerase chain reaction-restriction fragment length polymorphism analysis. Furthermore, plasma soluble PECAM-1 (sPECAM-1) levels were quantified by ELISA. The results of the present study demonstrated significantly higher genotype and allele frequencies of the Leu125Val polymorphism in PECAM-1 in the DVT group as compared with those in the control group (P<0.05). The plasma levels of sPECAM-1 in the DVT group (83.4 ± 23.5 ng/ml) were also significantly higher as compared with those in the control group (60.4 ± 19.4 ng/ml, P<0.01). In the patients with DVT, plasma levels of sPECAM-1 were significantly higher in those with the Leu/Val and Val/Val genotypes as compared with those possessing the Leu/Leu genotype (P<0.05). The PECAM-1 Leu125Val polymorphism was shown to be associated with an increased risk of DVT and PECAM-1 protein expression levels in venous vessels. In patients with DVT, the PECAM-1 Leu/Val and Val/Val genotypes were associated with delayed thrombus resolution, as determined by thrombus scoring, as compared with that in patients possessing the Leu/Val genotype. In conclusion, the present study indicated that PECAM-1 Leu125Val polymorphism and sPECAM-1 levels may be associated with DVT.

Asn563Ser polymorphism of CD31/PECAM-1 is associated with atherosclerotic cerebral infarction in a southern Han population.

BackgroundCD31, also called platelet endothelial cell adhesion molecule-1 (PECAM-1), is thought to play a role in the pathological mechanisms of atherosclerosis. Leu125Val polymorphism and elevated plasma levels of soluble PECAM-1 (sPECAM-1) were found to be associated with cerebral infarction. Our aim was to investigate the association between the Asn563Ser polymorphism of CD31/PECAM-1, plasma level of sPECAM-1, and the risk of atherosclerotic cerebral infarction (ACI) in the southern Han population of the People’s Republic of China.Subjects and methodsA total of 147 subjects with ACI and 114 controls were enrolled in the study. The Asn563Ser CD31/PECAM-1 polymorphism was detected using the polymerase chain reaction–restriction fragment length polymorphism method. The plasma spECAM-1 level was measured using the enzyme-linked immunosorbent assay method.ResultsIn this study, statistically significant differences in Asn563Ser genotype and allele distribution were found between the cases and controls (P<0.05). Furthermore, logistic regression analysis showed that the GG genotype is associated with increase in ACI risk (odds ratio =4.862, P<0.001). The plasma level of sPECAM-1 was associated with ACI (odds ratio =1.431, P=0.038). In both the ACI and the control groups, the plasma sPECAM-1 level in subjects with the GG genotype was higher than that in subjects carrying the AA or GA genotype (P<0.05).ConclusionOur study showed that the Asn563Ser polymorphism of CD31/PECAM-1 gene and elevated plasma sPECAM-1 level are related to ACI risk in the southern Han population of People’s Republic of China.