Circulating Extracellular Vesicle Proteins and MicroRNA Profiles in Subcortical and Cortical-Subcortical Ischaemic Stroke.

Laura Otero-Ortega,Exuperio Díez-Tejedor,Elisa Conde-Moreno,Susana B Bravo,Elisa Alonso-López,Luke Diekhorst,Mari Carmen Gómez-De Frutos,María Alonso De Leciñana,María Gutiérrez-Fernández,Fernando Laso-García,María Laura García-Bermejo,Blanca Fuentes,María Pérez-Mato

doi:10.3390/biomedicines9070786

Abstract

In order to investigate the role of circulating extracellular vesicles (EVs), proteins, and microRNAs as damage and repair markers in ischaemic stroke depending on its topography, subcortical (SC), and cortical-subcortical (CSC) involvement, we quantified the total amount of EVs using an enzyme-linked immunosorbent assay technique and analysed their global protein content using proteomics. We also employed a polymerase chain reaction to evaluate the circulating microRNA profile. The study included 81 patients with ischaemic stroke (26 SC and 55 CSC) and 22 healthy controls (HCs). No differences were found in circulating EV levels between the SC, CSC, and HC groups. We detected the specific expression of C1QA and Casp14 in the EVs of patients with CSC ischaemic stroke and the specific expression of ANXA2 in the EVs of patients with SC involvement. Patients with CSC ischaemic stroke showed a lower expression of miR-15a, miR-424, miR-100, and miR-339 compared with those with SC ischaemic stroke, and the levels of miR-339, miR-100, miR-199a, miR-369a, miR-424, and miR-15a were lower than those of the HCs. Circulating EV proteins and microRNAs from patients with CSC ischaemic stroke could be considered markers of neurite outgrowth, neurogenesis, inflammation process, and atherosclerosis. On the other hand, EV proteins and microRNAs from patients with SC ischaemic stroke might be markers of an anti-inflammatory process and blood–brain barrier disruption reduction.

Highlights

Extracellular vesicles (EVs) are membrane particles secreted by most cells that constitute major vehicles for intercellular communication
We determined that Apolipoprotein B (APOB), C3, A2M, C4BPA, and fibronectin 1 (FN1) were abundant proteins in the healthy controls (HCs), while APOB, C3, A2M, Von Willebrand factor (VWF), and FN1 were abundant in the SC group, and APOB, C3, A2M, C4BPA, and VWF were abundant in the CSC group
The levels of the proteins fibrinogen G (FGG), C4BPA, VWF, and FN1 were higher in the CSC group than in the HCs, and the ORM1 levels were higher in the SC group compared with the HCs

Summary

Introduction

Extracellular vesicles (EVs) are membrane particles secreted by most cells that constitute major vehicles for intercellular communication. In response to a stroke, EVs are released into the blood from brain cells and other organs [1,2,3,4,5]. These EVs can carry proteins and miRNAs that reveal important information regarding damage, protection, and the repair process after brain ischaemia, as well as stroke outcomes [6,7,8,9,10]. Most research studies on stroke have focused on the cortical neuronal ischaemic cascade but have ignored the subcortical damage with the involvement of white matter fibre and oligodendrocytes. Studying the subtypes of stroke that affect different locations, such as subcortical (SC) and cortical-subcortical (CSC)

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