CD31+ Extracellular Vesicles From Patients With Type 2 Diabetes Shuttle a miRNA Signature Associated With Cardiovascular Complications

Abstract

Innovative biomarkers are needed to improve the management of patients with type 2 diabetes mellitus (T2DM). Blood circulating miRNAs have been proposed as a potential tool to detect T2DM complications, but the lack of tissue specificity, among other reasons, has hampered their translation to clinical settings. Extracellular vesicle (EV)-shuttled miRNAs have been proposed as an alternative approach. Here, we adapted an immunomagnetic bead–based method to isolate plasma CD31+ EVs to harvest vesicles deriving from tissues relevant for T2DM complications. Surface marker characterization showed that CD31+ EVs were also positive for a range of markers typical of both platelets and activated endothelial cells. After characterization, we quantified 11 candidate miRNAs associated with vascular performance and shuttled by CD31+ EVs in a large (n = 218) cross-sectional cohort of patients categorized as having T2DM without complications, having T2DM with complications, and control subjects. We found that 10 of the tested miRNAs are affected by T2DM, while the signature composed by miR-146a, -320a, -422a, and -451a efficiently identified T2DM patients with complications. Furthermore, another CD31+ EV-shuttled miRNA signature, i.e., miR-155, -320a, -342-3p, -376, and -422a, detected T2DM patients with a previous major adverse cardiovascular event. Many of these miRNAs significantly correlate with clinical variables held to play a key role in the development of complications. In addition, we show that CD31+ EVs from patients with T2DM are able to promote the expression of selected inflammatory mRNAs, i.e., CCL2, IL-1α, and TNFα, when administered to endothelial cells in vitro. Overall, these data suggest that the miRNA cargo of plasma CD31+ EVs is largely affected by T2DM and related complications, encouraging further research to explore the diagnostic potential and the functional role of these alterations.

MicroRNA 146a is associated with diabetic complications in type 1 diabetic patients from the EURODIAB PCS

“…The source of serum miR-146a-5p was not established in our study; however, endothelial cells, platelets, and immune/inflammatory cells are likely candidates [ 36 , 37 ]. In these cell types, miR-146a-5p expression can be modulated by diabetes.…”

Role of endothelial miR-24 in COVID-19 cerebrovascular events

“…Levels of microRNAs (miRNAs) within extracellular vesicles (EVs) have been shown to be useful diagnostic and prognostic biomarkers in a number of disease states [ 1 – 3 ]. However, EVs miRNAs have never been investigated in COVID-19.…”

“…A SARS-CoV-2 test (RT-qPCR) was performed in all subjects to confirm or rule out the COVID-19 diagnosis. EC-EVs were extracted from the plasma collected from these patients via serial centrifugation and CD31 + magnetic isolation [ 1 ], and EC-EVs miR-24 levels were quantified as described [ 1 , 4 , 6 ].…”

Expression Profiles of Circulating microRNAs in South African Type 2 Diabetic Individuals on Treatment

“…In a total of 1989 participants that were enrolled in the VMH study, 1,254 met the inclusion criteria for this study, including 92 screen-detected diabetes, 188 known diabetics on treatment and 974 normotolerant individuals. Similarly, a recently conducted cross-sectional investigation, sourced their sample from a much larger, previously reported cohort, with a total sample size of 218 test subjects ( Prattichizzo et al, 2021a ). With ours being larger, this then validates the size of the chosen sample in this study.…”