The Association of Glucose Control with Circulating Levels of Red Blood Cell-Derived Vesicles in Type 2 Diabetes Mellitus Patients with Atrial Fibrillation

Abstract

Hyperglycemia is a trigger for structural alteration of red blood cells (RBCs) and their ability to release extracellular vesicles (EVs). The aim of the study was to elucidate whether glucose control in T2DM patients with concomitant HF and AF affects a circulating number of RBC-derived EVs. We prospectively included 417 T2DM patients with HF, 51 of them had atrial fibrillation and 25 healthy volunteers and 30 T2DM non-HF individuals. Clinical assessment, echocardiography examination and biomarker measures were performed at the baseline of the study. RBC-derived EVs were determined as CD235a+ PS+ particles by flow cytometry. NT-proBNP levels were measured by ELISA. AF patients with glycosylated hemoglobin (HbA1c) < 6.9% had lower levels of CD235a+ PS+ RBC-derived vesicles than those with HbA1c ≥ 7.0%. There were no significant differences in number of CD235a+ PS+ RBC-derived vesicles between patients in entire cohort and in non-AF sub-cohort with HbA1c < 6.9% and HbA1c ≥ 7.0%, respectively. Multivariate linear regression yielded that CD235a+ PS+ RBC-derived vesicles ≥ 545 particles in µL (OR = 1.06; 95% CI = 1.01−1.11, p = 0.044) independently predicted HbA1c ≥ 7.0%. Elevated levels of CD235a+ PS+ RBC-derived EVs independently predicted poor glycaemia control in T2DM patients with HF and AF.

Keywords: diabetes mellitus; extracellular vesicles; heart failure; hyperglycemia; red blood cells.

Figure 1

Amount of circulating CD235a+ PS+ RBC-derived vesicles: (Left panel) depicts results of flow cytometry measure of PS+ CD235a+ RBS-derived vesicles; Distinguishing populations of RBC-derived vesicles were determined at the first step of gaiting depending on their size (<0.5 μm) using forward scatter and side scatter dot plot (A). Next, CD235a+ RBC- derived vesicles and PS+ RBC- derived vesicles were determined as double-positive events for labels indicated as CD235a+ and PS+ in T2DM non-HF (B), T2DM patients with HF (C) when compared with healthy volunteers (D). (Right panel) illustrate comparable proportion of circulating amount of CD235a+ PS+ RBC-derived vesicles in different cohorts of eligible individuals. Abbreviations: T2DM, type 2 diabetes mellitus; HF, heart failure; AF, atrial fibrillation, RBC, red blood cells; PS, phosphatidylserine.

Figure 2

The amount of CD235a+ PS+ RBC-derived vesicles in T2DM patients with HF depending of glycemia control. Abbreviations: T2DM, type 2 diabetes mellitus; HF, heart failure; AF, atrial fibrillation, RBC, red blood cells; PS, phosphatidylserine; HbA1c, glycosilated hemoglobin.

Figure 3

The predictive ability of circulating amount of CD235a+ PS+ RBC-derived vesicles for poor glycaemia control The Receive Operation Characteristics curve analysis. Abbreviations: AUC, area under curve; CI, confidence interval; RBC, red blood cells; PS, phosphatidylserine.

Figure 4

Study design and patients’ procedure flow chart. Abbreviations: AF, atrial fibrillation; CABG, coronary artery bypass grafting; CV, cardiovascular; CKD, chronic kidney disease; GFR, glomerular filtration rate; HF, heart failure; HbA1c, glycated hemoglobin; HOMA-IR, Homeostatic Assessment Model of Insulin Resistance; N-terminal brain natriuretic pro-peptide; T2DM, type 2 diabetes mellitus; TIA, transient ischemic attack.