Extracellular microvesicle micro<scp>RNA</scp>s in children with sickle cell anaemia with divergent clinical phenotypes

Khalyfa, Ahamed; Akbarpour, Mahzad; Connes, Philippe; Romana, Marc; Lapping-Carr, Gabrielle; Zhang, Chunyu; Andrade, Jorge; Gozal, David

doi:10.1111/bjh.14104

Cited by 40 publications

(52 citation statements)

References 65 publications

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“…Indeed, we have recently shown that exosomes isolated from SCA patients impacted the endothelial cells phenotype and induced monocyte adhesion in a severity dependent manner [53]. We have also identified signature of exosomal microRNAs that distinguished severe from mild SCA patients [53]. Our present data do not provide evidence that large MPs are biomarkers of the three SCD complications analyzed.…”

Section: Discussioncontrasting

confidence: 64%

“…It should be pointed-out that our flow cytometer could not detect small MPs (diameter < 400 nm), which may be relevant bio-markers and bio-effectors. Indeed, we have recently shown that exosomes isolated from SCA patients impacted the endothelial cells phenotype and induced monocyte adhesion in a severity dependent manner [53]. We have also identified signature of exosomal microRNAs that distinguished severe from mild SCA patients [53].…”

Section: Discussionmentioning

confidence: 99%

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Differences of microparticle patterns between sickle cell anemia and hemoglobin SC patients

et al. 2017

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Sickle cell anemia (SCA) and hemoglobin SC (HbSC) disease are the two most common forms of sickle cell disease (SCD), a frequent hemoglobinopathy which exhibits a highly variable clinical course. Although high levels of microparticles (MPs) have been consistently reported in SCA and evidence of their harmful impact on the SCA complication occurrences have been provided, no data on MP pattern in HbSC patients has been reported so far. In this study, we determined and compared the MP patterns of 84 HbSC and 96 SCA children, all at steady-state, using flow cytometry. Most of circulating MPs were derived from platelets (PLTs) and red blood cells (RBCs) in the two SCD syndromes. Moreover, we showed that HbSC patients exhibited lower blood concentration of total MPs compared to SCA patients, resulting mainly from a decrease of MP levels originated from RBCs and to a lesser extent from PLTs. We did not detect any association between blood MP concentrations and the occurrence of painful vaso-occlusive crises, acute chest syndrome and pulmonary hypertension in both patient groups. We also demonstrated for the first time, that whatever the considered genotype, RBC-derived MPs exhibited higher externalized phosphatidylserine level and were larger than PLT-derived MPs.

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Section: Discussioncontrasting

confidence: 64%

Section: Discussionmentioning

confidence: 99%

Differences of microparticle patterns between sickle cell anemia and hemoglobin SC patients

et al. 2017

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“…The supernatants were aspirated and discarded, and the exosome pellets were re-suspended in 1× PBS buffer and then stored at −80°C until used. Exosome size was determined using electron microscopy as previously described [52]. …”

Section: Methodsmentioning

confidence: 99%

Circulating exosomes potentiate tumor malignant properties in a mouse model of chronic sleep fragmentation

et al. 2016

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BackgroundChronic sleep fragmentation (SF) increases cancer aggressiveness in mice. Exosomes exhibit pleiotropic biological functions, including immune regulatory functions, antigen presentation, intracellular communication and inter-cellular transfer of RNA and proteins. We hypothesized that SF-induced alterations in biosynthesis and cargo of plasma exosomes may affect tumor cell properties.ResultsSF-derived exosomes increased tumor cell proliferation (~13%), migration (~2.3-fold) and extravasation (~10%) when compared to exosomes from SC-exposed mice. Similarly, Pre exosomes from OSA patients significantly enhanced proliferation and migration of human adenocarcinoma cells compared to Post. SF-exosomal cargo revealed 3 discrete differentially expressed miRNAs, and exploration of potential mRNA targets in TC1 tumor cells uncovered 132 differentially expressed genes that encode for multiple cancer-related pathways.MethodsPlasma-derived exosomes from C57/B6 mice exposed to 6 wks of SF or sleep control (SC), and from adult human patients with obstructive sleep apnea (OSA) before (Pre) and after adherent treatment for 6 wks (Post) were co-cultured with mouse lung TC1 or human adenocarcinoma tumor cell lines, respectively. Proliferation, migration, invasion, endothelial barrier integrity and extravasation assays of tumor cells were performed. Plasma mouse exosomal miRNAs were profiled with arrays, and transcriptomic assessments of TC1 cells exposed to SF or SC exosomes were conducted to identify gene targets.ConclusionsChronic SF induces alterations in exosomal miRNA cargo that alter the biological properties of TC1 lung tumor cells to enhance their proliferative, migratory and extravasation properties, and similar findings occur in OSA patients, in whom SF is a constitutive component of their sleep disorder. Thus, exosomes could participate, at least in part, in the adverse cancer outcomes observed in OSA.

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“…28,29 Exosomes were isolated from plasma using the Total Exosome Isolation Kit according to the manufacturer's protocol (Life Technologies, Carlsbad, CA, USA), and further characterized as previously reported. 30,31 Briefly, plasma was centrifuged at 2000×g for 20 min to remove cell debris. The supernatants were collected and 0.2 volume of the Total Exosome Isolation Reagent was added.…”

Section: Exosome Isolation Labeling and Characterizationmentioning

confidence: 99%

“…30,31 For this, 15-μL drops of isolated exosomes in Dulbecco phosphate buffered saline (DPBS) were placed on parafilm before the Formvar/carboncoated grids are placed on top of exosome drops and allowed to stand for 5 to 10 min for exosome adsorption. Grids with adherent exosomes were transferred to three 30-μL drops of DPBS for washing, fixed with 2% paraformaldehyde in DPBS for 7 min, then incubated with 25-μL drops of 2% uranyl acetate and examined by electron microscopy.…”

Section: Electron Microscopymentioning

confidence: 99%

Effect on Intermittent Hypoxia on Plasma Exosomal Micro RNA Signature and Endothelial Function in Healthy Adults

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Zhang

Khalyfa

et al. 2016

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Study Objective: Intermittent hypoxia (IH) is associated with increased risk of cardiovascular disease. Exosomes are secreted by most cell types and released in biological fluids, including plasma, and play a role in modifying the functional phenotype of target cells. Using an experimental human model of IH, we investigated potential exosome-derived biomarkers of IH-induced vascular dysfunction. Methods: Ten male volunteers were exposed to room air (D0), IH (6 h/day) for 4 days (D4) and allowed to recover for 4 days (D8). Circulating plasma exosomes were isolated and incubated with human endothelial monolayer cultures for impedance measurements and RNA extracted and processed with messenger RNA (mRNA) arrays to identify gene targets. In addition, immunofluorescent assessments of endothelial nitric oxide synthase (eNOS) mRNA expression, ICAM-1 cellular distribution were conducted. Results: Plasma exosomal micro RNAs (miRNAs) were profiled. D4 exosomes, primarily from endothelial sources, disrupted impedance levels compared to D0 and D8. ICAM-1 expression was markedly upregulated in endothelial cells exposed to D4 exosomes along with significant reductions in eNOS expression. Microarray approaches identified a restricted and further validated signature of exosomal miRNAs in D4 exosomes, and mRNA arrays revealed putative endothelial gene target pathways. Conclusions:In humans, intermittent hypoxia alters exosome cargo in the circulation which promotes increased permeability and dysfunction of endothelial cells in vitro. A select number of circulating exosomal miRNAs may play important roles in the cardiovascular dysfunction associated with OSA by targeting specific effector pathways.

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Extracellular microvesicle microRNAs in children with sickle cell anaemia with divergent clinical phenotypes

Cited by 40 publications

References 65 publications

Differences of microparticle patterns between sickle cell anemia and hemoglobin SC patients

Differences of microparticle patterns between sickle cell anemia and hemoglobin SC patients

Circulating exosomes potentiate tumor malignant properties in a mouse model of chronic sleep fragmentation

Effect on Intermittent Hypoxia on Plasma Exosomal Micro RNA Signature and Endothelial Function in Healthy Adults

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