Background Circulating MicroRNAs (miRNAs) carried by microvesicles (MVs) have various physiologic... more 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 wit...
Glioblastoma (GB) is the most aggressive and recurrent form of brain cancer in adults. We hypothe... more Glioblastoma (GB) is the most aggressive and recurrent form of brain cancer in adults. We hypothesized that the identification of biomarkers such as certain microRNAs (miRNAs) and the circulating microvesicles (MVs) that transport them could be key to establishing GB progression, recurrence and therapeutic response. For this purpose, circulating MVs were isolated from the plasma of GB patients (before and after surgery) and of healthy subjects and characterized by flow cytometry. OpenArray profiling and the individual quantification of selected miRNAs in plasma and MVs was performed, followed by target genes’ prediction and in silico survival analysis. It was found that MVs’ parameters (number, EGFRvIII and EpCAM) decreased after the surgical resection of GB tumors, but the inter-patient variability was high. The expression of miR-106b-5p, miR-486-3p, miR-766-3p and miR-30d-5p in GB patients’ MVs was restored to control-like levels after surgery: miR-106b-5p, miR-486-3p and miR-766-...
Both cardiovascular disease and cancer continue to be causes of morbidity and mortality all over ... more Both cardiovascular disease and cancer continue to be causes of morbidity and mortality all over the world. Preventing and treating heart disease in patients undergoing cancer treatment remain an important and ongoing challenge for improving the lives of cancer patients, but also for their survival. Despite ongoing efforts to improve patient survival, minimal advances have been made in the early detection of cardiovascular disease in patients suffering from cancer. Understanding the communication between cancer and cardiovascular disease can be based on a deeper knowledge of the molecular mechanisms that define the profile of the bilateral network and establish disease-specific biomarkers and therapeutic targets. The role of exosomes, microvesicles, and apoptotic bodies, together defined as extracellular vesicles (EVs), in cross talk between cardiovascular disease and cancer is in an incipient form of research. Here, we will discuss the preclinical evidence on the bilateral connecti...
Mechanisms of Vascular Defects in Diabetes Mellitus, 2017
Diabetes mellitus, a metabolic disorder characterized by high blood glucose, is one of the main r... more Diabetes mellitus, a metabolic disorder characterized by high blood glucose, is one of the main risk factors in the development of vascular complications affecting both large and small blood vessels. A major challenge is the discovery of new mediators and biomarkers for diabetes-related vascular complications. In this regard, accumulating evidence indicate that endothelial progenitor cells (EPCs), derived from the bone marrow and peripheral blood, are critical for the maintenance and regeneration of endothelial cells contributing to repair and restoration of vascular wall integrity. The studies reveal that the reduced number of circulating EPCs under diabetic conditions can predict cardiovascular outcomes, and EPC dysfunction could contribute to the pathogenesis of diabetes – associated vascular disease.
Cardiac pathological hypertrophy is the major risk factor that usually progresses to heart failur... more Cardiac pathological hypertrophy is the major risk factor that usually progresses to heart failure. We hypothesized that extracellular vesicles (EVs), known to act as important mediators in regulating physiological and pathological functions, could have the potential to reduce the cardiac hypertrophy and the ensuing cardiovascular diseases. Herein, the effects of mesenchymal stem cell-derived extracellular vesicles (EV-MSCs) on cardiac hypertrophy were investigated. EVs were isolated from the secretome of human adipose tissue-derived stem cells (EV-ADSCs) or bone marrow-derived stem cells (EV-BMMSCs). Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were stimulated with AngII and TGF-β1, in absence or presence of EVs. The results showed that exposure of hiPSC-CMs to AngII and TGF-β1 generated in vitro model of hypertrophic cardiomyocytes characterized by increases in surface area, reactive oxygen species production, protein expression of cardiac-specific biomar...
Atherosclerosis is a progressive, chronic inflammatory disease of the large arteries caused by th... more Atherosclerosis is a progressive, chronic inflammatory disease of the large arteries caused by the constant accumulation of cholesterol, followed by endothelial dysfunction and vascular inflammation. We hypothesized that delivery of extracellular vesicles (EVs), recognized for their potential as therapeutic targets and tools, could restore vascular function in atherosclerosis. We explored by comparison the potential beneficial effects of EVs from subcutaneous adipose tissue stem cells (EVs (ADSCs)) or bone marrow mesenchymal stem cells (EVs (MSCs)) on the consequences of atherogenic diet on vascular health. Also, the influences of siRNA-targeting Smad2/3 (Smad2/3siRNA) on endothelial dysfunction and its key molecular players were analyzed. For this study, an animal model of atherosclerosis (HH) was transplanted with EVs (ADSCs) or EVs (MSCs) transfected or not with Smad2/3siRNA. For controls, healthy or HH animals were used. The results indicated that by comparison with the HH group...
Supplemental material, Supplementary_figure_1 for Diabetes-induced early molecular and functional... more Supplemental material, Supplementary_figure_1 for Diabetes-induced early molecular and functional changes in aortic heart valves in a murine model of atherosclerosis by Monica Madalina Tucureanu, Alexandru Filippi, Nicoleta Alexandru, Cristina Ana Constantinescu, Letitia Ciortan, Razvan Macarie, Mihaela Vadana, Geanina Voicu, Sabina Frunza, Dan Nistor, Agneta Simionescu, Dan Teodor Simionescu, Adriana Georgescu and Ileana Manduteanu in Diabetes & Vascular Disease Research
Thrombin-activatable fibrinolysis inhibitor (TAFI) plays an important role in coagulation and fib... more Thrombin-activatable fibrinolysis inhibitor (TAFI) plays an important role in coagulation and fibrinolysis. Whereas TAFI deficiency may lead to a haemorrhagic tendency, data from TAFI knockout mice (TAFI-/-) are controversial and no differences have been reported in these animals after ischemic stroke. There are also no data regarding the role of circulating microparticles (MPs) in TAFI-/-. to examine the effect of tPA on the rate of intracranial haemorrhage (ICH) and on MPs generated in a model of ischemic stroke in TAFI-/- mice. Thrombin was injected into the middle cerebral artery (MCA) to analyse the effect of tPA (10mg/Kg) on the infarct size and haemorrhage in the absence of TAFI. Immunofluorescence for Fluoro-Jade C was performed on frozen brain slides to analyse neuronal degeneration after ischemia. MPs were isolated from mouse blood and their concentrations calculated by flow cytometry. Compared with saline, tPA significantly increased the infarct size in TAFI-/- mice (p<0.05). Although plasma fibrinolytic activity (fibrin plate assay) was higher in these animals, no macroscopic or microscopic ICH was detected. A positive signal for apoptosis and degenerating neurons was observed in the infarct area, being significantly higher in tPA treated TAFI-/- mice (p<0.05). Interestingly, higher numbers of MPs were found in TAFI-/- plasma as compared to wild type, after stroke (p<0.05). TAFI deficiency results in increased brain damage in a model of thrombolysis after ischemic stroke, which was not associated with bleeding but with neuronal degeneration and MP production.
Extracellular vesicles (EVs) are small anuclear vesicles, delimited by a lipid bilayer, released ... more Extracellular vesicles (EVs) are small anuclear vesicles, delimited by a lipid bilayer, released by almost all cell types, carrying functionally active biological molecules that can be transferred to the neighbouring or distant cells, inducing phenotypical and functional changes, relevant in various physio-pathological conditions. The microRNAs are the most significant active components transported by EVs, with crucial role in intercellular communication and significant effects on recipient cells. They may also server as novel valuable biomarkers for the diagnosis of metabolic disorders. Moreover, EVs are supposed to mediate type 2 diabetes mellitus (T2DM) risk and its progress. The T2DM development is preceded by prediabetes, a state that is associated with early forms of nephropathy and neuropathy, chronic kidney disease, diabetic retinopathy, and increased risk of macrovascular disease. Although the interest of scientists was focused not only on the pathogenesis of diabetes, but also on the early diagnosis, little is known about EVs-incorporated microRNA involvement in prediabetes state and its microvascular and macrovascular complications. Here, we survey the biogenesis, classification, content, biological functions and the most popular primary isolation methods of EVs, review the EVs—associated microRNA profiling connexion with early stages of diabetes and discuss the role of EVs containing specific microRNAs in prediabetes complications.
Glioblastoma (GB) is the most aggressive form of brain cancer in adults, characterized by poor su... more Glioblastoma (GB) is the most aggressive form of brain cancer in adults, characterized by poor survival rates and lack of effective therapies. MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression post-transcriptionally through specific pairing with target messenger RNAs (mRNAs). Extracellular vesicles (EVs), a heterogeneous group of cell-derived vesicles, transport miRNAs, mRNAs and intracellular proteins, and have been shown to promote horizontal malignancy into adjacent tissue, as well as resistance to conventional therapies. Furthermore, GB-derived EVs have distinct miRNA contents and are able to penetrate the blood–brain barrier. Numerous studies have attempted to identify EV-associated miRNA biomarkers in serum/plasma and cerebrospinal fluid, but their collective findings fail to identify reliable biomarkers that can be applied in clinical settings. However, EVs carrying specific miRNAs or miRNA inhibitors have great potential as therapeutic nanotools in ...
Endothelial progenitor cells (EPCs) and circulating microparticles (MPs) have been proposed as ma... more Endothelial progenitor cells (EPCs) and circulating microparticles (MPs) have been proposed as markers of endothelial function. In turn, the protective role of irbesartan, an angiotensin receptor blocker, on the endothelium has been demonstrated in the recent years but the mechanism has not been yet fully elucidated. In this context, we investigated whether irbesartan can influence these two new biomarkers in patients with cardiovascular (CV) risk factors. We compared the levels of EPCs and MPs in patients with hypertension and dyslipidemia which received irbesartan with the levels in patients with the same CV risk factors but receiving other antihypertensive drugs and in healthy individuals. Using this methodology we showed the positive effect of angiotensin receptor blockade on the two proposed markers of endothelial function, reflecting both endothelial injury and repair, MPs and EPC respectively, in the same subjects with cardiovascular risk factors. Moreover, we found that pati...
INTRODUCTION Extracellular vesicles (EVs), including circulating microvesicles (MVs) or mi- cropa... more INTRODUCTION Extracellular vesicles (EVs), including circulating microvesicles (MVs) or mi- croparticles (MPs) and exosomes, derived from cells or platelets are present in the peripheral blood and are important elements involved in the activation of the coagulation system, transport of macromolecules and intercellular communication. In patients with vascular complications (including diabetes), the number of EVs is significantly increased during the acute phase of the disease. However, less is known about EVs release in the chronic state of diabetes. OBJECTIVES To analyse the profile of inflammatory cytokines and angiogenic factors in EVs in diabetic patients with ocular and vascular complications. PATIENTS AND METHODS The study included patients with diabetes and varying degrees of ocular complications including retinopathy (n = 48) and the control group (n = 13). EV-enriched and EV-depleted fractions were obtained from platelet-poor plasma by means of the centrifugation method (16 ...
Biochemical and biophysical research communications, Jan 25, 2016
The recognition of the importance of diabetes in vascular disease has greatly increased lately. C... more The recognition of the importance of diabetes in vascular disease has greatly increased lately. Common risk factors for diabetes-related vascular disease include hyperglycemia, insulin resistance, dyslipidemia, inflammation, hypercoagulability, hypertension, and atherosclerosis. All of these factors contribute to the endothelial dysfunction which generates the diabetic complications, both macro and microvascular. Knowledge of diabetes-related vascular complications and of associated mechanisms it is becoming increasingly important for therapists. The discovery of microparticles (MPs) and their associated microRNAs (miRNAs) have opened new perspectives capturing the attention of basic and clinical scientists for their potential to become new therapeutic targets and clinical biomarkers. MPs known as submicron vesicles generated from membranes of apoptotic or activated cells into circulation have the ability to act as autocrine and paracrine effectors in cell-to-cell communication. The...
s / Atherosclerosis 263 (2017) e29ee110 e48 effect on proMMP-9 activity. The peptides, did not sh... more s / Atherosclerosis 263 (2017) e29ee110 e48 effect on proMMP-9 activity. The peptides, did not show any effects on MMP mRNA levels. Conclusions: These data show that these peptides show a promising inhibitory effect on MMP-9 and MMP-2 activity in endothelial cells and macrophages. SAG050. ENDOTHELIAL PROGENITOR CELLS DERIVED MICROPARTICLES REPRODUCE THE FAVORABLE ROLE OF THEIR PARENT CELLS OF HEALTHY ORIGINS IN THE TREATMENT OF ATHEROSCLEROSIS VIA MICRORNA TRANSFER Adriana Georgescu, Nicoleta Alexandru, Emanuel Dragan, Florentina Safciuc, Ana Maria Daraban, Elisabeta Badila. 1 Institute of Cellular Biology and Pathology, Nicolae Simionescu of Romanian Academy, Bucharest, Romania; Carol Davila University of Medicine and Pharmacy, Bucharest, Romania; 3 Internal Medicine Clinic, Emergency Clinical Hospital, Bucharest, Romania Aim: Cell-derived microparticles (MPs), endothelial progenitor cells (EPCs) and microRNAs (miRNAs) have attracted major interest as biomarkers and potential regulators for atherosclerosis. We evaluated the possible protective role of EPC-derived MPs (MPEs) of healthy origins in atherosclerosis development as well as the mechanisms responsible for their repair capacity. Methods: The experiments were performed on hamsters divided into: (1) simultaneously hypertensive hyperlipidemic (HH) by combining two feeding conditions for 4 months; (2) HH with retro-orbital sinus injection containing MPEs, from control hamster, one doze per month for 4 months of HH diet, to prevent atherosclerosis (HHMPE); (3) controls (C), agematched normal healthy animals. Results: The results showed that: (1) MPE transplantation suppresses the development of atherosclerosis processes via: (i) the alleviation of dyslipidemia, hypertension, circulating EPC levels, cytokine/chemokine profiles (VEGF, IL-6, IL-8); (ii) the structural and functional remodeling within the vessel wall and heart; (2) MPEs operate as protective and delivery system for miRNAs in circulation; (3) MPEs protect against atherosclerotic vascular disease via transfer of miR-21, miR-126, miR-146a, mir-223 to circulating late EPCs. Conclusions: The data indicate that MPE transplantation can counteract HH diet-induced detrimental effects by their miRNA transfer to circulating EPCs mediating their function. These findings using MPEs as therapeutic tools for transferring miRNAs in an atherosclerotic animal model give hope to patients with cardiovascular disease. Acknowledgements: This work was supported by grants of the Romanian National Authority for Scientific Research, CNCSeUEFISCDI, project number PN-II-RU-TE-2014-4-0525 and PN-II-RU-TE-2014-4-0523, and by the Romanian Academy. SAG051. A CONDITIONAL TRANSGENIC MOUSE MODEL EXPRESSING APOE SPECIFICALLY IN THE ENDOTHELIUM Violeta Anca Gafencu, Ioana Madalina Fenyo, Madalina Dumitrescu, Elena Valeria Fuior, Ana Maria Eftimie, Violeta Georgeta Trusca, Camelia Stancu. Institute of Cellular Biology and Pathology Nicolae Simionescu,
Cells convey information among one another. One instrument employed to transmit data and constitu... more Cells convey information among one another. One instrument employed to transmit data and constituents to specific (target) cells is extracellular vesicles (EVs). They originate from a variety of cells (endothelial, immune cells, platelets, mesenchymal stromal cells, etc.), and consequently, their surface characteristics and cargo vary according to the paternal cell. The cargo could be DNA, mRNA, microRNA, receptors, metabolites, cytoplasmic proteins, or pathological molecules, as a function of which EVs exert different effects upon endocytosis in recipient cells. Recently, EVs have become important participants in a variety of pathologies, including atherogenesis and coronavirus disease 2019 (COVID-19)-associated thrombosis. Herein, we summarize recent advances and some of our own results on the role of EVs in atherosclerotic cardiovascular diseases, and discuss their potential to function as signaling mediators, biomarkers and therapeutic agents. Since COVID-19 patients have a high...
Diabetes reduces the number and induces dysfunction in circulating endothelial progenitor cells (... more Diabetes reduces the number and induces dysfunction in circulating endothelial progenitor cells (EPCs) by mechanisms that are still uncovered. This study aims to evaluate the number, viability, phenotype, and function of EPCs in dyslipidemic mice with early diabetes mellitus and EPC infiltration in the aortic valve in order to identify possible therapeutic targets in diabetes-associated cardiovascular disease. A streptozotocin-induced diabetic apolipoprotein E knock-out (ApoE−/−) mouse model was used to identify the early and progressive changes, at 4 or 7 days on atherogenic diet after the last streptozotocin or citrate buffer injection. Blood and aortic valves from diabetic or nondiabetic ApoE−/− animals were collected. EPCs were identified as CD34 and vascular endothelial growth factor receptor 2 positive monocytes, and the expression levels of α4β1, αVβ3, αVβ5, β1, αLβ2, α5 integrins, and C-X-C chemokine receptor type 4 chemokine receptor on EPC surface were assessed by flow cyt...
Background Circulating MicroRNAs (miRNAs) carried by microvesicles (MVs) have various physiologic... more 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 wit...
Glioblastoma (GB) is the most aggressive and recurrent form of brain cancer in adults. We hypothe... more Glioblastoma (GB) is the most aggressive and recurrent form of brain cancer in adults. We hypothesized that the identification of biomarkers such as certain microRNAs (miRNAs) and the circulating microvesicles (MVs) that transport them could be key to establishing GB progression, recurrence and therapeutic response. For this purpose, circulating MVs were isolated from the plasma of GB patients (before and after surgery) and of healthy subjects and characterized by flow cytometry. OpenArray profiling and the individual quantification of selected miRNAs in plasma and MVs was performed, followed by target genes’ prediction and in silico survival analysis. It was found that MVs’ parameters (number, EGFRvIII and EpCAM) decreased after the surgical resection of GB tumors, but the inter-patient variability was high. The expression of miR-106b-5p, miR-486-3p, miR-766-3p and miR-30d-5p in GB patients’ MVs was restored to control-like levels after surgery: miR-106b-5p, miR-486-3p and miR-766-...
Both cardiovascular disease and cancer continue to be causes of morbidity and mortality all over ... more Both cardiovascular disease and cancer continue to be causes of morbidity and mortality all over the world. Preventing and treating heart disease in patients undergoing cancer treatment remain an important and ongoing challenge for improving the lives of cancer patients, but also for their survival. Despite ongoing efforts to improve patient survival, minimal advances have been made in the early detection of cardiovascular disease in patients suffering from cancer. Understanding the communication between cancer and cardiovascular disease can be based on a deeper knowledge of the molecular mechanisms that define the profile of the bilateral network and establish disease-specific biomarkers and therapeutic targets. The role of exosomes, microvesicles, and apoptotic bodies, together defined as extracellular vesicles (EVs), in cross talk between cardiovascular disease and cancer is in an incipient form of research. Here, we will discuss the preclinical evidence on the bilateral connecti...
Mechanisms of Vascular Defects in Diabetes Mellitus, 2017
Diabetes mellitus, a metabolic disorder characterized by high blood glucose, is one of the main r... more Diabetes mellitus, a metabolic disorder characterized by high blood glucose, is one of the main risk factors in the development of vascular complications affecting both large and small blood vessels. A major challenge is the discovery of new mediators and biomarkers for diabetes-related vascular complications. In this regard, accumulating evidence indicate that endothelial progenitor cells (EPCs), derived from the bone marrow and peripheral blood, are critical for the maintenance and regeneration of endothelial cells contributing to repair and restoration of vascular wall integrity. The studies reveal that the reduced number of circulating EPCs under diabetic conditions can predict cardiovascular outcomes, and EPC dysfunction could contribute to the pathogenesis of diabetes – associated vascular disease.
Cardiac pathological hypertrophy is the major risk factor that usually progresses to heart failur... more Cardiac pathological hypertrophy is the major risk factor that usually progresses to heart failure. We hypothesized that extracellular vesicles (EVs), known to act as important mediators in regulating physiological and pathological functions, could have the potential to reduce the cardiac hypertrophy and the ensuing cardiovascular diseases. Herein, the effects of mesenchymal stem cell-derived extracellular vesicles (EV-MSCs) on cardiac hypertrophy were investigated. EVs were isolated from the secretome of human adipose tissue-derived stem cells (EV-ADSCs) or bone marrow-derived stem cells (EV-BMMSCs). Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were stimulated with AngII and TGF-β1, in absence or presence of EVs. The results showed that exposure of hiPSC-CMs to AngII and TGF-β1 generated in vitro model of hypertrophic cardiomyocytes characterized by increases in surface area, reactive oxygen species production, protein expression of cardiac-specific biomar...
Atherosclerosis is a progressive, chronic inflammatory disease of the large arteries caused by th... more Atherosclerosis is a progressive, chronic inflammatory disease of the large arteries caused by the constant accumulation of cholesterol, followed by endothelial dysfunction and vascular inflammation. We hypothesized that delivery of extracellular vesicles (EVs), recognized for their potential as therapeutic targets and tools, could restore vascular function in atherosclerosis. We explored by comparison the potential beneficial effects of EVs from subcutaneous adipose tissue stem cells (EVs (ADSCs)) or bone marrow mesenchymal stem cells (EVs (MSCs)) on the consequences of atherogenic diet on vascular health. Also, the influences of siRNA-targeting Smad2/3 (Smad2/3siRNA) on endothelial dysfunction and its key molecular players were analyzed. For this study, an animal model of atherosclerosis (HH) was transplanted with EVs (ADSCs) or EVs (MSCs) transfected or not with Smad2/3siRNA. For controls, healthy or HH animals were used. The results indicated that by comparison with the HH group...
Supplemental material, Supplementary_figure_1 for Diabetes-induced early molecular and functional... more Supplemental material, Supplementary_figure_1 for Diabetes-induced early molecular and functional changes in aortic heart valves in a murine model of atherosclerosis by Monica Madalina Tucureanu, Alexandru Filippi, Nicoleta Alexandru, Cristina Ana Constantinescu, Letitia Ciortan, Razvan Macarie, Mihaela Vadana, Geanina Voicu, Sabina Frunza, Dan Nistor, Agneta Simionescu, Dan Teodor Simionescu, Adriana Georgescu and Ileana Manduteanu in Diabetes & Vascular Disease Research
Thrombin-activatable fibrinolysis inhibitor (TAFI) plays an important role in coagulation and fib... more Thrombin-activatable fibrinolysis inhibitor (TAFI) plays an important role in coagulation and fibrinolysis. Whereas TAFI deficiency may lead to a haemorrhagic tendency, data from TAFI knockout mice (TAFI-/-) are controversial and no differences have been reported in these animals after ischemic stroke. There are also no data regarding the role of circulating microparticles (MPs) in TAFI-/-. to examine the effect of tPA on the rate of intracranial haemorrhage (ICH) and on MPs generated in a model of ischemic stroke in TAFI-/- mice. Thrombin was injected into the middle cerebral artery (MCA) to analyse the effect of tPA (10mg/Kg) on the infarct size and haemorrhage in the absence of TAFI. Immunofluorescence for Fluoro-Jade C was performed on frozen brain slides to analyse neuronal degeneration after ischemia. MPs were isolated from mouse blood and their concentrations calculated by flow cytometry. Compared with saline, tPA significantly increased the infarct size in TAFI-/- mice (p<0.05). Although plasma fibrinolytic activity (fibrin plate assay) was higher in these animals, no macroscopic or microscopic ICH was detected. A positive signal for apoptosis and degenerating neurons was observed in the infarct area, being significantly higher in tPA treated TAFI-/- mice (p<0.05). Interestingly, higher numbers of MPs were found in TAFI-/- plasma as compared to wild type, after stroke (p<0.05). TAFI deficiency results in increased brain damage in a model of thrombolysis after ischemic stroke, which was not associated with bleeding but with neuronal degeneration and MP production.
Extracellular vesicles (EVs) are small anuclear vesicles, delimited by a lipid bilayer, released ... more Extracellular vesicles (EVs) are small anuclear vesicles, delimited by a lipid bilayer, released by almost all cell types, carrying functionally active biological molecules that can be transferred to the neighbouring or distant cells, inducing phenotypical and functional changes, relevant in various physio-pathological conditions. The microRNAs are the most significant active components transported by EVs, with crucial role in intercellular communication and significant effects on recipient cells. They may also server as novel valuable biomarkers for the diagnosis of metabolic disorders. Moreover, EVs are supposed to mediate type 2 diabetes mellitus (T2DM) risk and its progress. The T2DM development is preceded by prediabetes, a state that is associated with early forms of nephropathy and neuropathy, chronic kidney disease, diabetic retinopathy, and increased risk of macrovascular disease. Although the interest of scientists was focused not only on the pathogenesis of diabetes, but also on the early diagnosis, little is known about EVs-incorporated microRNA involvement in prediabetes state and its microvascular and macrovascular complications. Here, we survey the biogenesis, classification, content, biological functions and the most popular primary isolation methods of EVs, review the EVs—associated microRNA profiling connexion with early stages of diabetes and discuss the role of EVs containing specific microRNAs in prediabetes complications.
Glioblastoma (GB) is the most aggressive form of brain cancer in adults, characterized by poor su... more Glioblastoma (GB) is the most aggressive form of brain cancer in adults, characterized by poor survival rates and lack of effective therapies. MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression post-transcriptionally through specific pairing with target messenger RNAs (mRNAs). Extracellular vesicles (EVs), a heterogeneous group of cell-derived vesicles, transport miRNAs, mRNAs and intracellular proteins, and have been shown to promote horizontal malignancy into adjacent tissue, as well as resistance to conventional therapies. Furthermore, GB-derived EVs have distinct miRNA contents and are able to penetrate the blood–brain barrier. Numerous studies have attempted to identify EV-associated miRNA biomarkers in serum/plasma and cerebrospinal fluid, but their collective findings fail to identify reliable biomarkers that can be applied in clinical settings. However, EVs carrying specific miRNAs or miRNA inhibitors have great potential as therapeutic nanotools in ...
Endothelial progenitor cells (EPCs) and circulating microparticles (MPs) have been proposed as ma... more Endothelial progenitor cells (EPCs) and circulating microparticles (MPs) have been proposed as markers of endothelial function. In turn, the protective role of irbesartan, an angiotensin receptor blocker, on the endothelium has been demonstrated in the recent years but the mechanism has not been yet fully elucidated. In this context, we investigated whether irbesartan can influence these two new biomarkers in patients with cardiovascular (CV) risk factors. We compared the levels of EPCs and MPs in patients with hypertension and dyslipidemia which received irbesartan with the levels in patients with the same CV risk factors but receiving other antihypertensive drugs and in healthy individuals. Using this methodology we showed the positive effect of angiotensin receptor blockade on the two proposed markers of endothelial function, reflecting both endothelial injury and repair, MPs and EPC respectively, in the same subjects with cardiovascular risk factors. Moreover, we found that pati...
INTRODUCTION Extracellular vesicles (EVs), including circulating microvesicles (MVs) or mi- cropa... more INTRODUCTION Extracellular vesicles (EVs), including circulating microvesicles (MVs) or mi- croparticles (MPs) and exosomes, derived from cells or platelets are present in the peripheral blood and are important elements involved in the activation of the coagulation system, transport of macromolecules and intercellular communication. In patients with vascular complications (including diabetes), the number of EVs is significantly increased during the acute phase of the disease. However, less is known about EVs release in the chronic state of diabetes. OBJECTIVES To analyse the profile of inflammatory cytokines and angiogenic factors in EVs in diabetic patients with ocular and vascular complications. PATIENTS AND METHODS The study included patients with diabetes and varying degrees of ocular complications including retinopathy (n = 48) and the control group (n = 13). EV-enriched and EV-depleted fractions were obtained from platelet-poor plasma by means of the centrifugation method (16 ...
Biochemical and biophysical research communications, Jan 25, 2016
The recognition of the importance of diabetes in vascular disease has greatly increased lately. C... more The recognition of the importance of diabetes in vascular disease has greatly increased lately. Common risk factors for diabetes-related vascular disease include hyperglycemia, insulin resistance, dyslipidemia, inflammation, hypercoagulability, hypertension, and atherosclerosis. All of these factors contribute to the endothelial dysfunction which generates the diabetic complications, both macro and microvascular. Knowledge of diabetes-related vascular complications and of associated mechanisms it is becoming increasingly important for therapists. The discovery of microparticles (MPs) and their associated microRNAs (miRNAs) have opened new perspectives capturing the attention of basic and clinical scientists for their potential to become new therapeutic targets and clinical biomarkers. MPs known as submicron vesicles generated from membranes of apoptotic or activated cells into circulation have the ability to act as autocrine and paracrine effectors in cell-to-cell communication. The...
s / Atherosclerosis 263 (2017) e29ee110 e48 effect on proMMP-9 activity. The peptides, did not sh... more s / Atherosclerosis 263 (2017) e29ee110 e48 effect on proMMP-9 activity. The peptides, did not show any effects on MMP mRNA levels. Conclusions: These data show that these peptides show a promising inhibitory effect on MMP-9 and MMP-2 activity in endothelial cells and macrophages. SAG050. ENDOTHELIAL PROGENITOR CELLS DERIVED MICROPARTICLES REPRODUCE THE FAVORABLE ROLE OF THEIR PARENT CELLS OF HEALTHY ORIGINS IN THE TREATMENT OF ATHEROSCLEROSIS VIA MICRORNA TRANSFER Adriana Georgescu, Nicoleta Alexandru, Emanuel Dragan, Florentina Safciuc, Ana Maria Daraban, Elisabeta Badila. 1 Institute of Cellular Biology and Pathology, Nicolae Simionescu of Romanian Academy, Bucharest, Romania; Carol Davila University of Medicine and Pharmacy, Bucharest, Romania; 3 Internal Medicine Clinic, Emergency Clinical Hospital, Bucharest, Romania Aim: Cell-derived microparticles (MPs), endothelial progenitor cells (EPCs) and microRNAs (miRNAs) have attracted major interest as biomarkers and potential regulators for atherosclerosis. We evaluated the possible protective role of EPC-derived MPs (MPEs) of healthy origins in atherosclerosis development as well as the mechanisms responsible for their repair capacity. Methods: The experiments were performed on hamsters divided into: (1) simultaneously hypertensive hyperlipidemic (HH) by combining two feeding conditions for 4 months; (2) HH with retro-orbital sinus injection containing MPEs, from control hamster, one doze per month for 4 months of HH diet, to prevent atherosclerosis (HHMPE); (3) controls (C), agematched normal healthy animals. Results: The results showed that: (1) MPE transplantation suppresses the development of atherosclerosis processes via: (i) the alleviation of dyslipidemia, hypertension, circulating EPC levels, cytokine/chemokine profiles (VEGF, IL-6, IL-8); (ii) the structural and functional remodeling within the vessel wall and heart; (2) MPEs operate as protective and delivery system for miRNAs in circulation; (3) MPEs protect against atherosclerotic vascular disease via transfer of miR-21, miR-126, miR-146a, mir-223 to circulating late EPCs. Conclusions: The data indicate that MPE transplantation can counteract HH diet-induced detrimental effects by their miRNA transfer to circulating EPCs mediating their function. These findings using MPEs as therapeutic tools for transferring miRNAs in an atherosclerotic animal model give hope to patients with cardiovascular disease. Acknowledgements: This work was supported by grants of the Romanian National Authority for Scientific Research, CNCSeUEFISCDI, project number PN-II-RU-TE-2014-4-0525 and PN-II-RU-TE-2014-4-0523, and by the Romanian Academy. SAG051. A CONDITIONAL TRANSGENIC MOUSE MODEL EXPRESSING APOE SPECIFICALLY IN THE ENDOTHELIUM Violeta Anca Gafencu, Ioana Madalina Fenyo, Madalina Dumitrescu, Elena Valeria Fuior, Ana Maria Eftimie, Violeta Georgeta Trusca, Camelia Stancu. Institute of Cellular Biology and Pathology Nicolae Simionescu,
Cells convey information among one another. One instrument employed to transmit data and constitu... more Cells convey information among one another. One instrument employed to transmit data and constituents to specific (target) cells is extracellular vesicles (EVs). They originate from a variety of cells (endothelial, immune cells, platelets, mesenchymal stromal cells, etc.), and consequently, their surface characteristics and cargo vary according to the paternal cell. The cargo could be DNA, mRNA, microRNA, receptors, metabolites, cytoplasmic proteins, or pathological molecules, as a function of which EVs exert different effects upon endocytosis in recipient cells. Recently, EVs have become important participants in a variety of pathologies, including atherogenesis and coronavirus disease 2019 (COVID-19)-associated thrombosis. Herein, we summarize recent advances and some of our own results on the role of EVs in atherosclerotic cardiovascular diseases, and discuss their potential to function as signaling mediators, biomarkers and therapeutic agents. Since COVID-19 patients have a high...
Diabetes reduces the number and induces dysfunction in circulating endothelial progenitor cells (... more Diabetes reduces the number and induces dysfunction in circulating endothelial progenitor cells (EPCs) by mechanisms that are still uncovered. This study aims to evaluate the number, viability, phenotype, and function of EPCs in dyslipidemic mice with early diabetes mellitus and EPC infiltration in the aortic valve in order to identify possible therapeutic targets in diabetes-associated cardiovascular disease. A streptozotocin-induced diabetic apolipoprotein E knock-out (ApoE−/−) mouse model was used to identify the early and progressive changes, at 4 or 7 days on atherogenic diet after the last streptozotocin or citrate buffer injection. Blood and aortic valves from diabetic or nondiabetic ApoE−/− animals were collected. EPCs were identified as CD34 and vascular endothelial growth factor receptor 2 positive monocytes, and the expression levels of α4β1, αVβ3, αVβ5, β1, αLβ2, α5 integrins, and C-X-C chemokine receptor type 4 chemokine receptor on EPC surface were assessed by flow cyt...
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Papers by Adriana Georgescu