Online Hemodiafiltration Inhibits Inflammation-Related Endothelial Dysfunction and Vascular Calcification of Uremic Patients Modulating miR-223 Expression in Plasma Extracellular Vesicles

Abstract

Decreased inflammation and cardiovascular mortality are evident in patients with end-stage chronic kidney disease treated by online hemodiafiltration. Extracellular vesicles (EV) are mediators of cell-to-cell communication and contain different RNA types. This study investigated whether mixed online hemodiafiltration (mOL-HDF) beneficial effects associate with changes in the RNA content of plasma EV in chronic kidney disease patients. Thirty bicarbonate hemodialysis (BHD) patients were randomized 1:1 to continue BHD or switch to mOL-HDF. Concentration, size, and microRNA content of plasma EV were evaluated for 9 mo; we then studied EV effects on inflammation, angiogenesis, and apoptosis of endothelial cells (HUVEC) and on osteoblast mineralization of vascular smooth muscle cells (VSMC). mOL-HDF treatment reduced different inflammatory markers, including circulating CRP, IL-6, and NGAL. All hemodialysis patients showed higher plasma levels of endothelial-derived EV than healthy subjects, with no significant differences between BHD and mOL-HDF. However, BHD-derived EV had an increased expression of the proatherogenic miR-223 with respect to healthy subjects or mOL-HDF. Compared with EV from healthy subjects, those from hemodialysis patients reduced angiogenesis and increased HUVEC apoptosis and VSMC calcification; however, all these detrimental effects were reduced with mOL-HDF with respect to BHD. Cell transfection with miR-223 mimic or antagomiR proved the role of this microRNA in EVinduced HUVEC and VSMC dysfunction. The switch from BHD to mOL-HDF significantly reduced systemic inflammation and miR-223 expression in plasma EV, thus improving HUVEC angiogenesis and reducing VSMC calcification.

miR-130a and Tgfβ Content in Extracellular Vesicles Derived from the Serum of Subjects at High Cardiovascular Risk Predicts their In-Vivo Angiogenic Potential

“…miR-130a target(s) validation. Transfection of mimic-miR-130a (10 nM) was performed on ECs using HiPerfect Transfection method (Qiagen, Valencia, USA) 63 . Total RNA from untreated ECs, ECs transfected with mimic-miR-130a, treated with sEV with high (D17, D18, D20) or low miR-130a content (D1, D2, D4) was extracted using All-in-One Purification kit (Norgen, Thorold, ON Canada).…”

“…miR-Scramble (Scr) was used as transfection reference sample. The expression of miR-130a was evaluated by RT-PCR 63 . miR-130a targets were analyzed by Western Blot analysis.…”

“…miR-130a targets were analyzed by Western Blot analysis. Proteins were obtained using RIPA buffer, as previously described 63 . Anti-KDR (1:200, Santa Cruz), anti-ROCK1 (1:200, Abcam), and anti-HOXA5 (1:200, Abcam) antibody, together with anti-βactin, were used (1:200, Santa Cruz).…”

The Epigenetic Landscape of Vascular Calcification: An Integrative Perspective

“…On the other hand, Pickering et al reported that serum miR-26a, miR-34a, and miR-223 did not differ between those with and without VC progression [77], findings that are in contrast to those by Cavallari et al, who described that plasma miR-223 increased in those with ESRD under chronic dialysis [49]. It is likely that the numbers of subjects being tested, the timing of the assay for circulatory miRNAs, the controls used to derive miRNA data, and the biologic fluid assayed may account for the discrepancy we observed.…”

“…Alternatively, VSMCs with miR-223 over-expression had increased proliferation and migratory ability, while calcified VSMCs had significantly up-regulated miR-223 levels [18]. Cavallari et al also showed that uremic patients with VC had nearly 2-fold higher levels of plasma exosomal miR-223 compared to control groups [49], lending support to the potential agonistic effect for VC for miR-223. This difference in miR-223 effect on VC seems to depend on cell type and the clinical context being evaluated.…”