Sabrina LA SALVIA - Academia.edu (original) (raw)

Papers by Sabrina LA SALVIA

American Journal of Physiology-Endocrinology and Metabolism

Elevated extracellular vesicles (EVs) are associated with glucose dysmetabolism. However, the eff... more Elevated extracellular vesicles (EVs) are associated with glucose dysmetabolism. However, the effects of insulin on EVs and subsequent relationships with insulin sensitivity, substrate oxidation, and inflammation are unknown. We tested the hypothesis that insulin would lower EVs and relate to insulin action. Fifty-one sedentary adults (54.8 ± 1.0 yr; VO2peak: 22.1 ± 0.6 ml/kg/min) with metabolic syndrome (MetS) and obesity (36.4 ± 0.65 kg/m2) underwent a 2-hr euglycemic-hyperinsulinemic clamp (5 mmol/L; 40 mU/m2/min). Count and size (medium: 200 - 624 nm; larger: 625 - 1000 nm) for total particle count, endothelial- (CD105+), leukocyte- (CD45+), platelet- (CD41+), and tetraspanin- (TX+: CD9/CD81/CD63), as well as platelet endothelial cell adhesion molecule- (CD31+) derived EVs were determined before and following the clamp using Full Spectrum Profiling (FSPTM). Size and MESF (Molecules of Equivalent Soluble Fluorochrome) data were generated using FCMPASS Software. Fat and carbohydra...

The Journal of Physiology, 2022

KEY POINTS What is already known that led to the present work? Extracellular vesicles (EVs) are i... more KEY POINTS What is already known that led to the present work? Extracellular vesicles (EVs) are increased in states of obesity and may play a role in altered insulin sensitivity and blood pressure. Aerobic exercise decreases fasting EV concentrations the following day in adults with obesity. What does this paper add to existing knowledge? This study directly tested the effects of insulin on EVs and how a single bout of exercise impacts these responses. Together, these data highlight the positive effects of a single bout of exercise on fasting and insulin-stimulated EVs, with the latter relating to increased insulin sensitivity and decreased augmentation index. What is the importance of the results for body function in health and/or disease? These results support future research identifying EV as a mechanistic factor in glucose regulation and vascular function as well as clinical use of exercise to reduce cardiovascular disease risk. ABSTRACT Extracellular vesicles (EVs) are often elevated in obesity and may modulate disease risk. Although acute exercise reduces fasting EVs in adults with obesity, no data exist on insulin-mediated EV responses. This study evaluated the effects of exercise on EV responses to insulin in relation to vascular function. Ten (5M/5F) sedentary adults with obesity (34.3 ± 3.7 kg/m2 ) completed an evening control and acute exercise condition (70% VO2 max to expend 400 kcals). Following an overnight fast, participants underwent a 2-hr euglycemic-hyperinsulinemic clamp (90 mg/dl; 40 mU/m2 /min) to determine metabolic insulin sensitivity (M-value), phenotypes of medium- to large-sized EVs, and aortic waveform measures. Endothelial (CD105+, CD41-/CD31+), leukocyte (CD45+), platelet (CD41+, CD41+/31+), and tetraspanin-derived EVs (TX+), as well as platelet endothelial cell adhesion molecule (CD31+) were determined before and after the clamp using high resolution spectral flow cytometry. Although exercise did not alter fasting hemodynamics, it lowered augmentation index (AIx75, P = 0.024) and increased the M-value (P = 0.042). Further, exercise decreased all fasting EVs (P<0.01) and decreased insulin-stimulated TX+ (P = 0.060), CD31+ (P = 0.060), and CD41-/31+ (P = 0.045) compared to rest. Interestingly, greater insulin-stimulated decreases in CD41-/31+ were associated with reduced AIx75 during the clamp (r = 0.62, P = 0.059), while insulin-stimulated decreases in CD41+ (r = -0.68, P = 0.031), CD41+/31+ (r = -0.69, P = 0.262), TX+ (r = -0.66, P = 0.037), and CD31+ (r = -0.69, P = 0.028) correlated with M-Value following exercise. Thus, acute exercise may decreases fasting and insulin-stimulated medium- to large-size EVs in conjunction with improved M-value and AIx75. More research is needed to understand effects of exercise on EVs in the regulation of glucose homeostasis and vascular function. Abstract figure legend A single bout of exercise raises metabolic insulin sensitivity and reduces aortic waveform hemodynamics in relation to extracellular vesicles (EVs). This article is protected by copyright. All rights reserved.

Current Hypertension Reports, 2020

Hypertension affects about half of all Americans, yet in the vast majority of cases, the factors ... more Hypertension affects about half of all Americans, yet in the vast majority of cases, the factors causing the hypertension cannot be clearly delineated. Developing a more precise understanding of the molecular pathogenesis of HTN and its various phenotypes is therefore a pressing priority. Circulating and urinary extracellular vesicles (EVs) are potential novel candidates as biomarkers and bioactivators in HTN. EVs are a heterogeneous population of small membrane fragments shed from various cell types into various body fluids. As EVs carry protein, RNA, and lipids, they also play a role as effectors and novel cell-to-cell communicators. In this review, we discuss the diagnostic, functional, and regenerative role of EVs in essential HTN and focus on EV protein and RNA cargo as the most extensively studied EV cargo. The field of EVs in HTN is still a young one and earlier studies have not used the novel EV detection tools currently available. More rigor and transparency in EV research are needed. Current data suggest that EVs represent potential novel biomarkers in HTN. EVs correlate with HTN severity and possibly end-organ damage. However, it has yet to be discerned which specific subtype(s) of EV reflects best HTN pathophysiology. Evolving studies are also showing that EVs might be novel regulators in vascular and renal tubular function and also be therapeutic. RNA in EVs has been studied in the context of hypertension, largely in the form of studies of miRNA, which are reviewed herein. Beyond miRNAs, mRNA in urinary EVs changed in response to sodium loading in humans. EVs represent promising novel biomarkers and bioactivators in essential HTN. Novel tools are being developed to apply more rigor in EV research including more in vivo models and translation to humans.

American Journal of Physiology-Renal Physiology, 2020

Extracellular vesicles (EVs) are novel mediators of cell-to-cell communication and appear to medi... more Extracellular vesicles (EVs) are novel mediators of cell-to-cell communication and appear to mediate the pathogenesis of hypertension (HTN). However, the mechanisms underlying the involvement of EVs in HTN remain unclear. The adaptive and innate immune systems play an important role affecting the kidney and vasculature in animal models of HTN. Evolving evidence shows that immune cell-derived EVs can modulate the immune system in a paracrine fashion and therefore may mediate the effects of inflammation in the pathogenesis of HTN. Therefore, we aimed to understand if specific subtypes of leukocyte/immune cell-derived EVs are altered in essential HTN using an in vivo model of angiotensin II (ANG II)-induced HTN. After 4 wk of ANG II treatment, EVs were isolated from the blood and kidney. EV origin and counts were characterized with Imaging Flow Cytometry, antibody panels targeting platelets, endothelial cells, and leukocytes including B and T cells, monocytes, and neutrophils. Leukocyt...

OBJECTIVE Air pollution has been recently identified as a risk factor for multiple sclerosis. Aim... more OBJECTIVE Air pollution has been recently identified as a risk factor for multiple sclerosis. Aim of this study was to investigate the immunological mechanism underlying the clinical association between air pollution, namely exposure to particulate matter 10 (PM10), and inflammatory activity of multiple sclerosis (MS) METHODS Daily recording of PM10 was obtained by monitors depending on the residence of subjects. Expression of molecules involved in activation, adhesion and migration of T lymphocytes were tested by flow-cytometry in 57 MS patients. We next assessed in vitro the effect of PM10 on expression of C-C chemokine receptors 6 (CCR6) by peripheral blood mononuclear cells (PBMCs), on cytokine production by monocyte-derived dendritic cells (mdDC) and on T cell polarization in PBMC/mdDC mixed cultures. RESULTS We identified a significant correlation between mean PM10 levels and expression of CCR6 CD4 + T circulating cells in MS patients. This was paralleled by the observation in...

Scientific Reports, 2020

Urinary extracellular vesicles (uEVs) provide bio-markers for kidney and urogenital diseases. Cen... more Urinary extracellular vesicles (uEVs) provide bio-markers for kidney and urogenital diseases. Centrifugation is the most common method used to enrich uEVs. However, a majority of studies to date have focused on the ultracentrifugation pellet, potentially losing a novel source of important biomarkers that could be obtained at lower centrifugation. Thus, the aim of this study is to rigorously characterize for the first time uEVs in the low speed pellet and determine the minimal volume of urine required for proteomic analysis (≥9.0 mL urine) and gene ontology classification identified 75% of the protein as extracellular exosomes. Cryo-Transmission Electron Microscopy (≥3.0 mL urine) provided evidence of a heterogeneous population of EVs for size and morphology independent of uromodulin filaments. Western blot detected several specific uEV kidney and EV markers (≥4.5 mL urine per lane). microRNAs quantification by qPCR was possible with urine volume as low as 0.5 mL. Particle enumeratio...

Diabetes, 2018

Microparticles (MPs) are a novel mediator and/or biomarker of cardiovascular disease (CVD) and ty... more Microparticles (MPs) are a novel mediator and/or biomarker of cardiovascular disease (CVD) and type 2 diabetes. A high fat meal can elevate MP release in relation to CVD risk, but the effects of carbohydrates are unknown. This is clinically relevant as low-fat diets are advised for glycemic control. We tested the hypothesis that a 75g oral glucose tolerance test (OGTT) would promote changes in MPs linked to CVD risk. Twenty-five obese adults (Age: 52.4±3.2y, BMI: 32.5±1.2kg/m2) were screened for prediabetes using ADA criteria (75g OGTT and/or HbA1c). Eight were normal glucose tolerant (NGT) and 17 had prediabetes (PD). Body composition (bioelectrical impedance) was measured and arterial stiffness (augmentation index; AI), insulin and glucose were collected at 0 and 2 hour of the OGTT to assess CVD risk. Annexin V+ (AV+) and Annexin V- (AV-) total MPs and platelet MPs (CD31+/CD41+; CD41+) were also collected at these times and analyzed from fresh plasma via imaging flow cytometry. Th...

Hypertension, 2019

Extracellular vesicles (EVs) have been described as novel biomarkers and bioactivators in vascula... more Extracellular vesicles (EVs) have been described as novel biomarkers and bioactivators in vascular dysfunction in hypertension. However, the mechanism(s) by which EVs affect vascular function is unknown. To examine the effects of EVs on endothelial-dependent vasodilation (acetylcholine), we isolated circulating EVs from platelet-poor plasma using a low centrifugation speed (17 000 g ) and mesenteric resistance arteries from 12-week-old normotensive WKYs (Wistar-Kyoto rats) and SHRs (spontaneously hypertensive rats). Arteries were cannulated on a pressure myograph, and EVs were added to the vessel lumen and circulating bath. We found that circulating EVs from normotensive WKY reduced vasodilation of normotensive WKY arteries but had no effect on hypertensive SHR arteries. In contrast, EVs from hypertensive SHR failed to reduce vasodilation of arteries from both WKY and SHR. The restraining effect on vasodilation by EVs from normotensive WKY may be mediated by inhibition of eNOS (endo...

Medicine & Science in Sports & Exercise, 2019

Hypertension, 2019

Infiltration of macrophages and T cells in kidneys and vasculature may play a role in the develop... more Infiltration of macrophages and T cells in kidneys and vasculature may play a role in the development of hypertension (HTN) and extracellular vesicles (EVs) have emerged as novel cell-to-cell communicators in the pathogenesis HTN. Immune cell-derived EVs can reflect the pathophysiological activity of the parental cell and mediate immune stimulation or inhibition in paracrine fashion. We investigated subgroups of circulating leukocyte-derived EVs (LEVs) in angiotensin II (Ang II)-induced HTN with or without candesartan (C) or hydralazine, hydrochlorothiazide, and reserpine (HHR). Ang II was delivered via osmotic mini pumps (500 ng/kg/min; n=15) and with C (10 mg/kg/day, n=5) or HHR (30 mg/kg/day, n=5) in drinking water for 4 weeks to 129S6 mice. Systolic blood pressure (SBP) was measured daily by tail-cuff manometry. After 4 weeks, mice were euthanized and citrated blood collected. EV isolation was performed by differential centrifugation, followed by EV phenotyping with imaging flow...

Frontiers in Neuroscience, 2019

Nutrients, 2019

Although extracellular vesicles (EVs) are a novel biomediator of type 2 diabetes (T2D) and cardio... more Although extracellular vesicles (EVs) are a novel biomediator of type 2 diabetes (T2D) and cardiovascular disease (CVD), the effects of hyperglycemia on EVs in humans is unknown. We tested the hypothesis that a 75-g oral glucose tolerance test (OGTT) would promote changes in EVs in relation to CVD risk. Twenty-five obese adults (Age: 52.4 ± 3.2 year, BMI: 32.5 ± 1.2 kg/m2) were screened for normal glucose tolerance (NGT, n = 8) and prediabetes (PD, n = 17) using American Diabetes Association criteria (75 g OGTT and/or HbA1c). Body composition (bioelectrical impedance) and fitness (VO2peak) were measured. Arterial stiffness (augmentation index; AIx) was measured at 0, 60- and 120-min while insulin, glucose, and free fatty acids were evaluated every 30 min during the OGTT to assess CVD risk. Annexin V positive (AV+) and Annexin V negative (AV-) total EVs, platelet EVs (CD31+/CD41+; CD41+), leukocyte EVs (CD45+; CD45+/CD41−), platelet endothelial cell adhesion molecule (PECAM) (CD31+) ...

Disease models & mechanisms, Mar 26, 2018

In the past, amyotrophic lateral sclerosis (ALS) has been considered a 'neurocentric' dis... more In the past, amyotrophic lateral sclerosis (ALS) has been considered a 'neurocentric' disease; however, new evidence suggests that it should instead be looked at from a 'multisystemic' or 'non-neurocentric' point of view. From 2006, we focused on the study of non-neural cells: ALS patients' peripheral blood mononuclear cells (PMBCs) and lymphoblastoid cell lines (LCLs). Here, we characterize LCLs of sporadic ALS (sALS) and patients carrying , and mutations to identify an ALS biologically relevant molecular signature, and determine whether and how mutations differentially affect ALS-linked pathways. Although LCLs are different from motor neurons (MNs), in LCLs we found some features typical of degenerating MNs in ALS, i.e. protein aggregation and mitochondrial dysfunction. Moreover, different gene mutations have different effects on ALS cellular mechanisms. and mutations imbalance mitochondrial dynamism toward increased fusion, whereas sALS and mutations m...

Hypertension, 2018

There is emerging evidence that extracellular vesicles (EVs) may be novel bio-markers and bio-act... more There is emerging evidence that extracellular vesicles (EVs) may be novel bio-markers and bio-activators in the pathogenesis of HTN. We hypothesize that the phenotypes of EVs from the circulation and kidney are altered in Angiotensin II (AII) induced HTN and normalize with anti-hypertensive treatment (Rx). AII was delivered via osmotic mini pumps (500 ng/kg/min) alone, or with candesartan (C) @ 10 mg/kg/day, or with hydralazine, hydrochlorothiazide, reserpine (HHR) @ 30/10/0.2 mg/kg/day in drinking water for 2 weeks to 129S6 mice (n = 5 each). Systolic blood pressure (SBP) was measured daily for 2 weeks by tail-cuff manometry. After 2 weeks, mice were euthanized and citrated blood and kidneys collected. Kidney EVs (KEVs) were isolated after incubation with Collagenase type A and subsequent DC. Both circulating EVs and KEVs were isolated through differential centrifugation and characterized by imaging flow cytometry (AmnisImage-StreamX Mark II) using PECAM-1 (CD31), leukocyte (CD45),...

Frontiers in Neuroscience, 2018

Amyotrophic lateral sclerosis (ALS) is a progressive adult-onset neurodegenerative disease, that ... more Amyotrophic lateral sclerosis (ALS) is a progressive adult-onset neurodegenerative disease, that affects cortical, bulbar and spinal motor neurons, and it is considered a proteinopathy, in which pathological proteins (SOD1, TDP-43, and FUS) may accumulate and interfere with neuronal functions eventually leading to cell death. These proteins can be released from cells and transported in the body fluids by extracellular vesicles (EVs). EVs are spherical vesicles, which are classified mainly in microvesicles (MVs) and exosomes (EXOs) based on their biogenesis, size and surface markers. In this study we characterized MVs and EXOs isolated from plasma of sporadic ALS patients and healthy controls and determined their number, size and SOD1, TDP-43, and FUS protein composition. No variation was found in the number of EVs between ALS patients and controls. However, the mean size both for MVs and for EXOs resulted increased in ALS patients compared to controls. MVs derived from ALS patients were enriched in SOD1, TDP-43, phospho-TDP-43, and FUS proteins compared to CTRLs. SOD1 was generally more concentrated in EXOs than in MVs, while TDP-43 and FUS protein levels were slightly higher in MVs than in EXOs. We demonstrated that MVs and EXOs size were increased in ALS patients compared to controls and that MVs of ALS patients were enriched with toxic proteins compared to CTRLs. EXOs did not show any protein changes. These data may suggest that MVs can transport toxic proteins and might play a role in prion-like propagation of ALS disease.

Circulating extracellular vesicles (EVs) contain molecular footprints from their cell of origin a... more Circulating extracellular vesicles (EVs) contain molecular footprints from their cell of origin and may provide potential non-invasive access for detection, characterization, and monitoring of numerous diseases. Despite their growing promise, the integrated proteo-transcriptomic landscape of EVs and their donor cells remain poorly understood. To assess their cargo, we conducted small RNA sequencing and mass spectrometry (LC-MS/MS) of EVs isolated from in vitro cancer cell culture and prostate cancer patients’ serum. Here, we report that EVs enrich for distinct molecular cargo, and their proteo-transcriptome is predominantly different from their cancer cell of origin, implicating a coordinated disposal and delivery mechanism. We have discovered that EVs package their cargo in a non-random fusion, as their most enriched RNAs and proteins are not the most abundant cargo from their donor cells. We show that EVs enrich for 4 times more cytoskeletal and 2 times extracellular proteins than...

American Journal of Physiology-Endocrinology and Metabolism

Elevated extracellular vesicles (EVs) are associated with glucose dysmetabolism. However, the eff... more Elevated extracellular vesicles (EVs) are associated with glucose dysmetabolism. However, the effects of insulin on EVs and subsequent relationships with insulin sensitivity, substrate oxidation, and inflammation are unknown. We tested the hypothesis that insulin would lower EVs and relate to insulin action. Fifty-one sedentary adults (54.8 ± 1.0 yr; VO2peak: 22.1 ± 0.6 ml/kg/min) with metabolic syndrome (MetS) and obesity (36.4 ± 0.65 kg/m2) underwent a 2-hr euglycemic-hyperinsulinemic clamp (5 mmol/L; 40 mU/m2/min). Count and size (medium: 200 - 624 nm; larger: 625 - 1000 nm) for total particle count, endothelial- (CD105+), leukocyte- (CD45+), platelet- (CD41+), and tetraspanin- (TX+: CD9/CD81/CD63), as well as platelet endothelial cell adhesion molecule- (CD31+) derived EVs were determined before and following the clamp using Full Spectrum Profiling (FSPTM). Size and MESF (Molecules of Equivalent Soluble Fluorochrome) data were generated using FCMPASS Software. Fat and carbohydra...

The Journal of Physiology, 2022

KEY POINTS What is already known that led to the present work? Extracellular vesicles (EVs) are i... more KEY POINTS What is already known that led to the present work? Extracellular vesicles (EVs) are increased in states of obesity and may play a role in altered insulin sensitivity and blood pressure. Aerobic exercise decreases fasting EV concentrations the following day in adults with obesity. What does this paper add to existing knowledge? This study directly tested the effects of insulin on EVs and how a single bout of exercise impacts these responses. Together, these data highlight the positive effects of a single bout of exercise on fasting and insulin-stimulated EVs, with the latter relating to increased insulin sensitivity and decreased augmentation index. What is the importance of the results for body function in health and/or disease? These results support future research identifying EV as a mechanistic factor in glucose regulation and vascular function as well as clinical use of exercise to reduce cardiovascular disease risk. ABSTRACT Extracellular vesicles (EVs) are often elevated in obesity and may modulate disease risk. Although acute exercise reduces fasting EVs in adults with obesity, no data exist on insulin-mediated EV responses. This study evaluated the effects of exercise on EV responses to insulin in relation to vascular function. Ten (5M/5F) sedentary adults with obesity (34.3 ± 3.7 kg/m2 ) completed an evening control and acute exercise condition (70% VO2 max to expend 400 kcals). Following an overnight fast, participants underwent a 2-hr euglycemic-hyperinsulinemic clamp (90 mg/dl; 40 mU/m2 /min) to determine metabolic insulin sensitivity (M-value), phenotypes of medium- to large-sized EVs, and aortic waveform measures. Endothelial (CD105+, CD41-/CD31+), leukocyte (CD45+), platelet (CD41+, CD41+/31+), and tetraspanin-derived EVs (TX+), as well as platelet endothelial cell adhesion molecule (CD31+) were determined before and after the clamp using high resolution spectral flow cytometry. Although exercise did not alter fasting hemodynamics, it lowered augmentation index (AIx75, P = 0.024) and increased the M-value (P = 0.042). Further, exercise decreased all fasting EVs (P<0.01) and decreased insulin-stimulated TX+ (P = 0.060), CD31+ (P = 0.060), and CD41-/31+ (P = 0.045) compared to rest. Interestingly, greater insulin-stimulated decreases in CD41-/31+ were associated with reduced AIx75 during the clamp (r = 0.62, P = 0.059), while insulin-stimulated decreases in CD41+ (r = -0.68, P = 0.031), CD41+/31+ (r = -0.69, P = 0.262), TX+ (r = -0.66, P = 0.037), and CD31+ (r = -0.69, P = 0.028) correlated with M-Value following exercise. Thus, acute exercise may decreases fasting and insulin-stimulated medium- to large-size EVs in conjunction with improved M-value and AIx75. More research is needed to understand effects of exercise on EVs in the regulation of glucose homeostasis and vascular function. Abstract figure legend A single bout of exercise raises metabolic insulin sensitivity and reduces aortic waveform hemodynamics in relation to extracellular vesicles (EVs). This article is protected by copyright. All rights reserved.

Current Hypertension Reports, 2020

Hypertension affects about half of all Americans, yet in the vast majority of cases, the factors ... more Hypertension affects about half of all Americans, yet in the vast majority of cases, the factors causing the hypertension cannot be clearly delineated. Developing a more precise understanding of the molecular pathogenesis of HTN and its various phenotypes is therefore a pressing priority. Circulating and urinary extracellular vesicles (EVs) are potential novel candidates as biomarkers and bioactivators in HTN. EVs are a heterogeneous population of small membrane fragments shed from various cell types into various body fluids. As EVs carry protein, RNA, and lipids, they also play a role as effectors and novel cell-to-cell communicators. In this review, we discuss the diagnostic, functional, and regenerative role of EVs in essential HTN and focus on EV protein and RNA cargo as the most extensively studied EV cargo. The field of EVs in HTN is still a young one and earlier studies have not used the novel EV detection tools currently available. More rigor and transparency in EV research are needed. Current data suggest that EVs represent potential novel biomarkers in HTN. EVs correlate with HTN severity and possibly end-organ damage. However, it has yet to be discerned which specific subtype(s) of EV reflects best HTN pathophysiology. Evolving studies are also showing that EVs might be novel regulators in vascular and renal tubular function and also be therapeutic. RNA in EVs has been studied in the context of hypertension, largely in the form of studies of miRNA, which are reviewed herein. Beyond miRNAs, mRNA in urinary EVs changed in response to sodium loading in humans. EVs represent promising novel biomarkers and bioactivators in essential HTN. Novel tools are being developed to apply more rigor in EV research including more in vivo models and translation to humans.

American Journal of Physiology-Renal Physiology, 2020

Extracellular vesicles (EVs) are novel mediators of cell-to-cell communication and appear to medi... more Extracellular vesicles (EVs) are novel mediators of cell-to-cell communication and appear to mediate the pathogenesis of hypertension (HTN). However, the mechanisms underlying the involvement of EVs in HTN remain unclear. The adaptive and innate immune systems play an important role affecting the kidney and vasculature in animal models of HTN. Evolving evidence shows that immune cell-derived EVs can modulate the immune system in a paracrine fashion and therefore may mediate the effects of inflammation in the pathogenesis of HTN. Therefore, we aimed to understand if specific subtypes of leukocyte/immune cell-derived EVs are altered in essential HTN using an in vivo model of angiotensin II (ANG II)-induced HTN. After 4 wk of ANG II treatment, EVs were isolated from the blood and kidney. EV origin and counts were characterized with Imaging Flow Cytometry, antibody panels targeting platelets, endothelial cells, and leukocytes including B and T cells, monocytes, and neutrophils. Leukocyt...

OBJECTIVE Air pollution has been recently identified as a risk factor for multiple sclerosis. Aim... more OBJECTIVE Air pollution has been recently identified as a risk factor for multiple sclerosis. Aim of this study was to investigate the immunological mechanism underlying the clinical association between air pollution, namely exposure to particulate matter 10 (PM10), and inflammatory activity of multiple sclerosis (MS) METHODS Daily recording of PM10 was obtained by monitors depending on the residence of subjects. Expression of molecules involved in activation, adhesion and migration of T lymphocytes were tested by flow-cytometry in 57 MS patients. We next assessed in vitro the effect of PM10 on expression of C-C chemokine receptors 6 (CCR6) by peripheral blood mononuclear cells (PBMCs), on cytokine production by monocyte-derived dendritic cells (mdDC) and on T cell polarization in PBMC/mdDC mixed cultures. RESULTS We identified a significant correlation between mean PM10 levels and expression of CCR6 CD4 + T circulating cells in MS patients. This was paralleled by the observation in...

Scientific Reports, 2020

Urinary extracellular vesicles (uEVs) provide bio-markers for kidney and urogenital diseases. Cen... more Urinary extracellular vesicles (uEVs) provide bio-markers for kidney and urogenital diseases. Centrifugation is the most common method used to enrich uEVs. However, a majority of studies to date have focused on the ultracentrifugation pellet, potentially losing a novel source of important biomarkers that could be obtained at lower centrifugation. Thus, the aim of this study is to rigorously characterize for the first time uEVs in the low speed pellet and determine the minimal volume of urine required for proteomic analysis (≥9.0 mL urine) and gene ontology classification identified 75% of the protein as extracellular exosomes. Cryo-Transmission Electron Microscopy (≥3.0 mL urine) provided evidence of a heterogeneous population of EVs for size and morphology independent of uromodulin filaments. Western blot detected several specific uEV kidney and EV markers (≥4.5 mL urine per lane). microRNAs quantification by qPCR was possible with urine volume as low as 0.5 mL. Particle enumeratio...

Diabetes, 2018

Microparticles (MPs) are a novel mediator and/or biomarker of cardiovascular disease (CVD) and ty... more Microparticles (MPs) are a novel mediator and/or biomarker of cardiovascular disease (CVD) and type 2 diabetes. A high fat meal can elevate MP release in relation to CVD risk, but the effects of carbohydrates are unknown. This is clinically relevant as low-fat diets are advised for glycemic control. We tested the hypothesis that a 75g oral glucose tolerance test (OGTT) would promote changes in MPs linked to CVD risk. Twenty-five obese adults (Age: 52.4±3.2y, BMI: 32.5±1.2kg/m2) were screened for prediabetes using ADA criteria (75g OGTT and/or HbA1c). Eight were normal glucose tolerant (NGT) and 17 had prediabetes (PD). Body composition (bioelectrical impedance) was measured and arterial stiffness (augmentation index; AI), insulin and glucose were collected at 0 and 2 hour of the OGTT to assess CVD risk. Annexin V+ (AV+) and Annexin V- (AV-) total MPs and platelet MPs (CD31+/CD41+; CD41+) were also collected at these times and analyzed from fresh plasma via imaging flow cytometry. Th...

Hypertension, 2019

Extracellular vesicles (EVs) have been described as novel biomarkers and bioactivators in vascula... more Extracellular vesicles (EVs) have been described as novel biomarkers and bioactivators in vascular dysfunction in hypertension. However, the mechanism(s) by which EVs affect vascular function is unknown. To examine the effects of EVs on endothelial-dependent vasodilation (acetylcholine), we isolated circulating EVs from platelet-poor plasma using a low centrifugation speed (17 000 g ) and mesenteric resistance arteries from 12-week-old normotensive WKYs (Wistar-Kyoto rats) and SHRs (spontaneously hypertensive rats). Arteries were cannulated on a pressure myograph, and EVs were added to the vessel lumen and circulating bath. We found that circulating EVs from normotensive WKY reduced vasodilation of normotensive WKY arteries but had no effect on hypertensive SHR arteries. In contrast, EVs from hypertensive SHR failed to reduce vasodilation of arteries from both WKY and SHR. The restraining effect on vasodilation by EVs from normotensive WKY may be mediated by inhibition of eNOS (endo...

Medicine & Science in Sports & Exercise, 2019

Hypertension, 2019

Infiltration of macrophages and T cells in kidneys and vasculature may play a role in the develop... more Infiltration of macrophages and T cells in kidneys and vasculature may play a role in the development of hypertension (HTN) and extracellular vesicles (EVs) have emerged as novel cell-to-cell communicators in the pathogenesis HTN. Immune cell-derived EVs can reflect the pathophysiological activity of the parental cell and mediate immune stimulation or inhibition in paracrine fashion. We investigated subgroups of circulating leukocyte-derived EVs (LEVs) in angiotensin II (Ang II)-induced HTN with or without candesartan (C) or hydralazine, hydrochlorothiazide, and reserpine (HHR). Ang II was delivered via osmotic mini pumps (500 ng/kg/min; n=15) and with C (10 mg/kg/day, n=5) or HHR (30 mg/kg/day, n=5) in drinking water for 4 weeks to 129S6 mice. Systolic blood pressure (SBP) was measured daily by tail-cuff manometry. After 4 weeks, mice were euthanized and citrated blood collected. EV isolation was performed by differential centrifugation, followed by EV phenotyping with imaging flow...

Frontiers in Neuroscience, 2019

Nutrients, 2019

Although extracellular vesicles (EVs) are a novel biomediator of type 2 diabetes (T2D) and cardio... more Although extracellular vesicles (EVs) are a novel biomediator of type 2 diabetes (T2D) and cardiovascular disease (CVD), the effects of hyperglycemia on EVs in humans is unknown. We tested the hypothesis that a 75-g oral glucose tolerance test (OGTT) would promote changes in EVs in relation to CVD risk. Twenty-five obese adults (Age: 52.4 ± 3.2 year, BMI: 32.5 ± 1.2 kg/m2) were screened for normal glucose tolerance (NGT, n = 8) and prediabetes (PD, n = 17) using American Diabetes Association criteria (75 g OGTT and/or HbA1c). Body composition (bioelectrical impedance) and fitness (VO2peak) were measured. Arterial stiffness (augmentation index; AIx) was measured at 0, 60- and 120-min while insulin, glucose, and free fatty acids were evaluated every 30 min during the OGTT to assess CVD risk. Annexin V positive (AV+) and Annexin V negative (AV-) total EVs, platelet EVs (CD31+/CD41+; CD41+), leukocyte EVs (CD45+; CD45+/CD41−), platelet endothelial cell adhesion molecule (PECAM) (CD31+) ...

Disease models & mechanisms, Mar 26, 2018

In the past, amyotrophic lateral sclerosis (ALS) has been considered a 'neurocentric' dis... more In the past, amyotrophic lateral sclerosis (ALS) has been considered a 'neurocentric' disease; however, new evidence suggests that it should instead be looked at from a 'multisystemic' or 'non-neurocentric' point of view. From 2006, we focused on the study of non-neural cells: ALS patients' peripheral blood mononuclear cells (PMBCs) and lymphoblastoid cell lines (LCLs). Here, we characterize LCLs of sporadic ALS (sALS) and patients carrying , and mutations to identify an ALS biologically relevant molecular signature, and determine whether and how mutations differentially affect ALS-linked pathways. Although LCLs are different from motor neurons (MNs), in LCLs we found some features typical of degenerating MNs in ALS, i.e. protein aggregation and mitochondrial dysfunction. Moreover, different gene mutations have different effects on ALS cellular mechanisms. and mutations imbalance mitochondrial dynamism toward increased fusion, whereas sALS and mutations m...

Hypertension, 2018

There is emerging evidence that extracellular vesicles (EVs) may be novel bio-markers and bio-act... more There is emerging evidence that extracellular vesicles (EVs) may be novel bio-markers and bio-activators in the pathogenesis of HTN. We hypothesize that the phenotypes of EVs from the circulation and kidney are altered in Angiotensin II (AII) induced HTN and normalize with anti-hypertensive treatment (Rx). AII was delivered via osmotic mini pumps (500 ng/kg/min) alone, or with candesartan (C) @ 10 mg/kg/day, or with hydralazine, hydrochlorothiazide, reserpine (HHR) @ 30/10/0.2 mg/kg/day in drinking water for 2 weeks to 129S6 mice (n = 5 each). Systolic blood pressure (SBP) was measured daily for 2 weeks by tail-cuff manometry. After 2 weeks, mice were euthanized and citrated blood and kidneys collected. Kidney EVs (KEVs) were isolated after incubation with Collagenase type A and subsequent DC. Both circulating EVs and KEVs were isolated through differential centrifugation and characterized by imaging flow cytometry (AmnisImage-StreamX Mark II) using PECAM-1 (CD31), leukocyte (CD45),...

Frontiers in Neuroscience, 2018

Amyotrophic lateral sclerosis (ALS) is a progressive adult-onset neurodegenerative disease, that ... more Amyotrophic lateral sclerosis (ALS) is a progressive adult-onset neurodegenerative disease, that affects cortical, bulbar and spinal motor neurons, and it is considered a proteinopathy, in which pathological proteins (SOD1, TDP-43, and FUS) may accumulate and interfere with neuronal functions eventually leading to cell death. These proteins can be released from cells and transported in the body fluids by extracellular vesicles (EVs). EVs are spherical vesicles, which are classified mainly in microvesicles (MVs) and exosomes (EXOs) based on their biogenesis, size and surface markers. In this study we characterized MVs and EXOs isolated from plasma of sporadic ALS patients and healthy controls and determined their number, size and SOD1, TDP-43, and FUS protein composition. No variation was found in the number of EVs between ALS patients and controls. However, the mean size both for MVs and for EXOs resulted increased in ALS patients compared to controls. MVs derived from ALS patients were enriched in SOD1, TDP-43, phospho-TDP-43, and FUS proteins compared to CTRLs. SOD1 was generally more concentrated in EXOs than in MVs, while TDP-43 and FUS protein levels were slightly higher in MVs than in EXOs. We demonstrated that MVs and EXOs size were increased in ALS patients compared to controls and that MVs of ALS patients were enriched with toxic proteins compared to CTRLs. EXOs did not show any protein changes. These data may suggest that MVs can transport toxic proteins and might play a role in prion-like propagation of ALS disease.

Circulating extracellular vesicles (EVs) contain molecular footprints from their cell of origin a... more Circulating extracellular vesicles (EVs) contain molecular footprints from their cell of origin and may provide potential non-invasive access for detection, characterization, and monitoring of numerous diseases. Despite their growing promise, the integrated proteo-transcriptomic landscape of EVs and their donor cells remain poorly understood. To assess their cargo, we conducted small RNA sequencing and mass spectrometry (LC-MS/MS) of EVs isolated from in vitro cancer cell culture and prostate cancer patients’ serum. Here, we report that EVs enrich for distinct molecular cargo, and their proteo-transcriptome is predominantly different from their cancer cell of origin, implicating a coordinated disposal and delivery mechanism. We have discovered that EVs package their cargo in a non-random fusion, as their most enriched RNAs and proteins are not the most abundant cargo from their donor cells. We show that EVs enrich for 4 times more cytoskeletal and 2 times extracellular proteins than...