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High glucose changes the proteomic composition of eMPs. (a) Venn diagram depicts differences in protein composition among eMP populations. (b) Functional analysis (gene ontology biological processes and Reactome pathways) of the 68 eMPHG-exclusive proteins using the ClueGO plug-in for Cytoscape. Functional over-enrichments with p < 0.05 are presented
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Aims/hypothesis:
Individuals with diabetes exhibit increases in circulating endothelial microparticles (eMPs, also referred to as endothelial microvesicles), which are associated with endothelial dysfunction and a heightened risk of cardiovascular complications. We have shown that eMPs are markers and mediators of vascular injury although their ro...
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(1) Background: survivors of allogeneic hematopoietic cell transplantation (alloHCT) suffer from morbidity and mortality due to cardiovascular events. We hypothesized that vascular injury and pro-coagulant activity are evident in alloHCT survivors without existing alloHCT complications or relapse. (2) Methods: we enrolled consecutive adult alloHCT...
Citations
... Hence, thorough and systematic investigation could harness the usefulness of the individual surface markers for diagnosis, as specific surface markers could inform the function and disease status of ECs which is useful for uncovering mechanisms underlying KD (116,127,133). For instance, CD62E + EMP reveals that the ECs are in the activated state [129] Other research has shown that proteomics on EMPs from diseased states could reveal mechanisms underlying disease progression [148,149]. ...
Kawasaki disease is a febrile illness characterised by systemic inflammation of small- and medium-sized blood vessels which commonly occurs in young children. Although self-limiting, there is a risk of developing coronary artery lesions as disease progresses with delay in diagnosis and treatment. Unfortunately, the diagnosis of KD continues to remain a clinical dilemma. Thus, this article not only summarises the key research gaps associated to KD, but also evaluates the possibility of using circulating endothelial injury biomarkers such as, circulating endothelial cells, endothelial microparticles and vascular endothelial cell-free DNA, as diagnostic and prognostic tool of KD: a “liquid biopsy” approach. The challenges of translating liquid biopsies to use in KD, and the opportunities for improvement in its diagnosis and management that such translation may provide are discussed. The use of endothelial damage markers, which are easily obtained via blood collection, as diagnostics is promising and we hope this will be translated to clinical applications in the near future.
... Furthermore, we demonstrated that IL-1β contributed to the high glucose-induced apoptosis of RPEs. High glucose levels in cell culture media can lead to dysfunction of HUVECs [16], resulting in increased inflammatory responses and oxidative stress, ultimately leading to dysfunction of RPEs [17,18]. Our study showed that high glucose activated the NF-κB pathway in endothelial cells, which promoted the expression of inflammatory factors and accumulation of ROS, leading to endothelial cell apoptosis and release of IL-1β. ...
Objectives
Diabetic retinopathy (DR) is a retinal disease that arises from impaired glucose tolerance and leads to retinal microvascular leakages. Recent studies have indicated that DR pathogenesis is linked to dysfunctional retinal pigment epithelial (RPE) cells.
Methods
Investigating the potential interplay between endothelial cells (ECs) and RPE cells by treating ECs with high glucose (HG) and evaluating the function of cytokines released from ECs on the growth of RPE cells.
Results
The results revealed that high glucose-stimulated Human Umbilical Vein Endothelial Cells (HUVECs) activated the NF-κB signaling pathway, increased intracellular levels of reactive oxygen species (ROS) and expression of caspase 3 while also elevating HUVECs delivery of cytokines such as VEGF, TNF-α, IL-6, and IL-1β.
Conclusions
As a result of our study, cytokines released from HG-treated HUVECs impede the growth of ARPE-19 in vitro , highlighting the importance of functional ECs for exploring the underlying mechanisms of vascular-associated retinal dysfunction. Inflammatory factors secreted from endothelial cells induced by high glucose impair human retinal pigment epithelial cells.
... D-glucose is the major carbon source in the cell culture growth medium. Increased release of EVs has been observed in the presence of both elevated and reduced glucose levels, and this effect seems cell type-dependent (Burger et al., 2017;Garcia et al., 2015;Rice et al., 2015;Thom et al., 2017). Several reports have shown a change in the biochemical composition or biological function of EVs when producing cells cultured in a medium containing high concentrations of glucose (Davidson et al., 2018;De Jong et al., 2012;Huang et al., 2020;Lin et al., 2019;Thom et al., 2017;Wu et al., 2017;Zhu et al., 2019;Zhou et al., 2021). ...
... EV secretion is regulated in response to acute stimuli and in diseased states. Our data showing increased liver EV secretion in response to hyperglycaemia, both in isolated liver slices and in mice in vivo, extends on previous findings reporting nutrient-driven EV secretion in several cultured cell types including hepatocytes 49,50 , adipocytes 51 , monocytes 52 and endothelial cells 53 . While many factors probably regulate EV secretion, together these data demonstrate a mechanism whereby acute increases in blood glucose are sensed by the liver, which results in enhanced EV secretion. ...
Small extracellular vesicles (EVs) are signalling messengers that regulate inter-tissue communication through delivery of their molecular cargo. Here, we show that liver-derived EVs are acute regulators of whole-body glycaemic control in mice. Liver EV secretion into the circulation is increased in response to hyperglycaemia, resulting in increased glucose effectiveness and insulin secretion through direct inter-organ EV signalling to skeletal muscle and the pancreas, respectively. This acute blood glucose lowering effect occurs in healthy and obese mice with non-alcoholic fatty liver disease, despite marked remodelling of the liver-derived EV proteome in obese mice. The EV-mediated blood glucose lowering effects were recapitulated by administration of liver EVs derived from humans with or without progressive non-alcoholic fatty liver disease, suggesting broad functional conservation of liver EV signalling and potential therapeutic utility. Taken together, this work reveals a mechanism whereby liver EVs act on peripheral tissues via endocrine signalling to restore euglycaemia in the postprandial state.
... MVs express numerous features of cells of origin including specific surface antigens and receptors, so they were named according to the cells they come from, for example endothelial, platelet and leukocyte MVs [12]. In vitro studies confirmed that, high glucose concentration activates endothelial cells (ECs) to discharge significant amount of MVs, called endothelial microvesicles (EMVs), with a distinct molecular composition [13,14]. These endothelial membrane-derived vesicles are also reported to carry regulating molecules crucial in cellto-cell communication. ...
Background
Circulating MicroRNAs (miRNAs) carried by microvesicles (MVs) have various physiological and pathological functions by post-transcriptional regulation of gene expression being considered markers for many diseases including diabetes and dyslipidemia. We aimed to identify new common miRNAs both in MVs and plasma that could be predictive biomarkers for diabetic dyslipidemia evolution.
Methods
For this purpose, plasma from 63 participants in the study (17 type 2 diabetic patients, 17 patients with type 2 diabetes and dyslipidemia, 14 patients with dyslipidemia alone and 15 clinically healthy persons without diabetes or dyslipidemia) was used for the analysis of circulating cytokines, MVs, miRNAs and MV-associated miRNAs.
Results
The results uncovered three miRNAs, miR-218, miR-132 and miR-143, whose expression was found to be significantly up-regulated in both circulating MVs and plasma from diabetic patients with dyslipidemia. These miRNAs showed significant correlations with important plasma markers, representative of this pathology. Thus, MV/plasma miR-218 was negatively correlated with the levels of erythrocyte MVs, plasma miR-132 was positively connected with MV miR-132 and negatively with uric acid and erythrocyte plasma levels, and plasma miR-143 was negatively related with creatinine levels and diastolic blood pressure. Also, three miRNAs common to MV and plasma, namely miR-21, miR-122, and miR-155, were identified to be down-regulated and up-regulated, respectively, in diabetic dyslipidemia. In addition, MV miR-21 was positively linked with cholesterol plasma levels and plasma miR-21 with TNFα plasma levels, MV miR-122 was negatively correlated with LDL-c levels and plasma miR-122 with creatinine and diastolic blood pressure and positively with MV miR-126 levels, MV miR-155 was positively associated with cholesterol and total MV levels and negatively with HDL-c levels, whereas plasma miR-155 was positively correlated with Il-1β plasma levels and total MV levels and negatively with MV miR-223 levels.
Conclusions
In conclusion, miR-218, miR-132, miR-143, and miR-21, miR-122, miR-155 show potential as biomarkers for diabetic dyslipidemia, but there is a need for more in-depth studies. These findings bring new information regarding the molecular biomarkers specific to diabetic dyslipidemia and could have important implications for the treatment of patients affected by this pathology.
... EV elevation in plasma from patients with T2DM and prediabetes suggests that chronic hyperglycemia is related to EVs release; furthermore, high glucose levels increase EVs released from HUVEC cultures. [21] HOMA-IR high value is a hallmark of impaired glucose metabolism in T2DM patients; [22] in this study, HOMA-IR is lower in IFG than in T2DM patients but is not different from the control subjects. A remarkable difference between the normoglycemic subjects versus IFG and T2DM patients was the highest total cholesterol level in the latter two groups. ...
Background: The goal of this study was to analyze the in vitro effect of plasma-isolated extracellular vesicles (EVs) from patients with impaired fasting glucose (IFG) on cell migration and angiogenic score on human endothelial cells (EA.hy926) cultures. Methods: Plasma samples from five patients with IFG, five with Type 2 diabetes mellitus (T2DM), and five normoglycemic subjects (controls) were used. Plasma-derived EVs were characterized by electron microscopy, dynamic light scattering and Western blotting, vascular endothelial growth factor receptor 2 (VEGFR-2), and endoglin detected in EVs by flow cytometry; wound closure assays and angiogenic score by matrigel assays in EA.hy926 cells were performed. Results: EA.hy926 cell migration induced by plasma-derived EVs from patients with IFG was greater than in control subjects (P = 0.023). EVs from patients with T2DM and IFG induced higher angiogenic scores than EVs from control subjects (P = 0.012 and P = 0.036, respectively). Conclusions: Endoglin and VEGFR-2 levels in EVs from IFG or T2DM patients were not different from those in control subjects. Plasma-derived EVs from patients with IFG and T2DM positively influenced human endothelial cell migration and angiogenic activity in vitro.
... The effect of EVs released by inflamed endothelial cells (ECs) on the reprogramming of monocytes toward a proinflammatory phenotype has recently been studied [81]. In vitro studies on ECs exposed to hyperglycemic (HG) conditions demonstrated that the number and size of EVs change and that they have greater procoagulant activity, contributing to progressive endothelial injury [82]. Interestingly, the senescence similarly affected EVs' properties and miRNAs' cargo [83]. ...
A common denominator of metabolic diseases, including type 2 diabetes Mellitus, dyslipidemia, and atherosclerosis, are elevated oxidative stress and chronic inflammation. These complex, multi-factorial diseases are caused by the detrimental interaction between the individual genetic background and multiple environmental stimuli. The cells, including the endothelial ones, acquire a preactivated phenotype and metabolic memory, exhibiting increased oxidative stress, inflammatory gene expression, endothelial vascular activation, and prothrombotic events, leading to vascular complications. There are different pathways involved in the pathogenesis of metabolic diseases, and increased knowledge suggests a role of the activation of the NF-kB pathway and NLRP3 inflammasome as key mediators of metabolic inflammation. Epigenetic-wide associated studies provide new insight into the role of microRNAs in the phenomenon of metabolic memory and the development consequences of vessel damage. In this review, we will focus on the microRNAs related to the control of anti-oxidative enzymes, as well as microRNAs related to the control of mitochondrial functions and inflammation. The objective is the search for new therapeutic targets to improve the functioning of mitochondria and reduce oxidative stress and inflammation, despite the acquired metabolic memory.
... The impact of MVs on ROS is influenced by the target cell's circumstances and the cells that produce the vesicles. High-glucose circumstances have been demonstrated to generate a threefold increase in MV synthesis in endothelial cells, especially human umbilical vein endothelial cells (HUVEC), with variations in the molecular makeup of these vesicles [83]. In contrast, neutrophils produce different MVs in response to various activators [84]. ...
Microvesicles are cellular membrane vesicles of which size is limited to 30–1000 nm. Almost all cells release them in response to activation signals and apoptosis. Their ability for intercellular communication and enhancement of potential for information exchange (between them) has attracted much interest. Their content is affected by the content of the mother cell, which can help identify their origin. Furthermore, these particles can change the physiology of the target cells by transferring a set of molecules to them and changing the epigenetics of the cells by transferring DNA and RNA. These changes can be induced in cells close to the mother and distant cells. Significant activities of these microvesicles are known both in physiological and pathologic conditions. In this regard, we have reviewed these small particle elements, their contents, and the way of synthesis. Finally, we discussed their current known roles to reveal more potential applications in leukemia.
... Digested peptides were then analyzed by label-free LC-MS/MS through the OHRI Proteomics Core Facility, as described previously [63]. Briefly, the system consisted of an Ul-tiMate 3000 RSLC nano HPLC, LTQ Orbitrap XL hybrid mass spectrometer (ThermoFisher Scientific, Nepean, ON, Canada) the XCalibur software (version 2.0.7) and a nanospray ionization source. ...
Hypertension and diabetes induce vascular injury through processes that are not fully understood. Changes in extracellular vesicle (EV) composition could provide novel insights. Here, we examined the protein composition of circulating EVs from hypertensive, diabetic and healthy mice. EVs were isolated from transgenic mice overexpressing human renin in the liver (TtRhRen, hypertensive), OVE26 type 1 diabetic mice and wild-type (WT) mice. Protein content was analyzed using liquid chromatography–mass spectrometry. We identified 544 independent proteins, of which 408 were found in all groups, 34 were exclusive to WT, 16 were exclusive to OVE26 and 5 were exclusive to TTRhRen mice. Amongst the differentially expressed proteins, haptoglobin (HPT) was upregulated and ankyrin-1 (ANK1) was downregulated in OVE26 and TtRhRen mice compared with WT controls. Conversely, TSP4 and Co3A1 were upregulated and SAA4 was downregulated exclusively in diabetic mice; and PPN was upregulated and SPTB1 and SPTA1 were downregulated in hypertensive mice, compared to WT mice. Ingenuity pathway analysis identified enrichment in proteins associated with SNARE signaling, the complement system and NAD homeostasis in EVs from diabetic mice. Conversely, in EVs from hypertensive mice, there was enrichment in semaphroin and Rho signaling. Further analysis of these changes may improve understanding of vascular injury in hypertension and diabetes.
... Although no work exists in humans with obesity/MetS, in vitro work under hyperglycemic conditions show that platelet, leukocyte, and endothelial EVs impair acetylcholine mediated endothelial-dependent relaxation of carotid arteries and aorta, in part through, reduced eNOS and increased caveolin-1 (Ishida et al., 2016;Martin et al., 2004). Moreover, endothelial EVs incubated with hyperglycemia increase pro-coagulant activity ~300% and stimulate HUVEC ROS production by ~250% (Burger et al., 2017). However, others propose that EVs are modified by obesity to promote angiogenesis and improve heart function (Icli et al., 2013), suggesting that compensatory mechanisms in obesity/MetS may exist to minimize vascular dysfunction. ...
Metabolic Syndrome (MetS) raises cardiovascular disease risk. Extracellular vesicles (EVs) have emerged as important mediators of insulin sensitivity, although few studies on vascular function exist in humans. We determined the effect of insulin on EVs in relation to vascular function. Adults with MetS (n = 51, n = 9 M, 54.8 ± 1.0 years, 36.4 ± 0.7 kg/m2 , ATPIII: 3.5 ± 0.1 a.u., VO2 max: 22.1 ± 0.6 ml/kg/min) were enrolled in this cross-sectional study. Peripheral insulin sensitivity (M-value) was determined during a euglycemic clamp (40 mU/m2 /min, 90 mg/dl), and blood was collected for EVs (CD105+, CD45+, CD41+, TX+, and CD31+; spectral flow cytometry), inflammation, insulin, and substrates. Central hemodynamics (applanation tonometry) was determined at 0 and 120 min via aortic waveforms. Pressure myography was used to assess insulin-induced arterial vasodilation from mouse 3rd order mesenteric arteries (100-200 μm in diameter) at 0.2, 2 and 20 nM of insulin with EVs from healthy and MetS adults. Adults with MetS had low peripheral insulin sensitivity (2.6 ± 0.2 mg/kg/min) and high HOMA-IR (4.7 ± 0.4 a.u.) plus Adipose-IR (13.0 ± 1.3 a.u.). Insulin decreased total/particle counts (p < 0.001), CD45+ EVs (p = 0.002), AIx75 (p = 0.005) and Pb (p = 0.04), FFA (p < 0.001), total adiponectin (p = 0.006), ICAM (p = 0.002), and VCAM (p = 0.03). Higher M-value related to lower fasted total EVs (r = -0.40, p = 0.004) while higher Adipose-IR associated with higher fasted EVs (r = 0.42, p = 0.004) independent of VAT. Fasting CD105+ and CD45+ derived total EVs correlated with fasting AIx75 (r = 0.29, p < 0.05) and Pb (r = 0.30, p < 0.05). EVs from MetS participants blunted insulin-induced vasodilation in mesenteric arteries compared with increases from healthy controls across insulin doses (all p < 0.005). These data highlight EVs as potentially novel mediators of vascular insulin sensitivity and disease risk.
