NPC‐EXs Alleviate Endothelial Oxidative Stress and Dysfunction through the miR‐210 Downstream Nox2 and VEGFR2 Pathways

Abstract

We have demonstrated that neural progenitor cells (NPCs) protect endothelial cells (ECs) from oxidative stress. Since exosomes (EXs) can convey the benefit of parent cells through their carried microRNAs (miRs) and miR-210 is ubiquitously expressed with versatile functions, we investigated the role of miR-210 in the effects of NPC-EXs on oxidative stress and dysfunction in ECs. NPCs were transfected with control and miR-210 scramble/inhibitor/mimic to generate NPC-EXscon, NPC-EXssc, NPC-EXsanti-miR-210, and NPC-EXsmiR-210. The effects of various NPC-EXs on angiotensin II- (Ang II-) induced reactive oxygen species (ROS) overproduction, apoptosis, and dysfunction, as well as dysregulation of Nox2, ephrin A3, VEGF, and p-VEGFR2/VEGFR2 in ECs were evaluated. Results showed (1) Ang II-induced ROS overproduction, increase in apoptosis, and decrease in tube formation ability, accompanied with Nox2 upregulation and reduction of p-VEGFR2/VEGFR2 in ECs. (2) Compared to NPC-EXscon or NPC-EXssc, NPC-EXsanti-miR-210 were less whereas NPC-EXsmiR-210 were more effective on attenuating these detrimental effects induced by Ang II in ECs. (3) These effects of NPC-EXsanti-miR-210 and NPC-EXsmiR-210 were associated with the changes of miR-210, ephrin A3, VEGF, and p-VEGFR2/VEGFR2 ratio in ECs. Altogether, the protective effects of NPC-EXs on Ang II-induced endothelial injury through miR-210 which controls Nox2/ROS and VEGF/VEGFR2 signals were studied.

Loading MiR-210 in Endothelial Progenitor Cells Derived Exosomes Boosts Their Beneficial Effects on Hypoxia/Reoxygeneation-Injured Human Endothelial Cells via Protecting Mitochondrial Function

“…The NanoSight NS300 (Malvern Instruments, Malvern, UK) was used to analyze the size and CD63 expression of EXs as previously described [27,28]. To detect CD63 expression in the EXs, the collected EXs were incubated with CD63-conjugated microbeads (10 ul; Miltenyi Biotec) in a 100-µl reaction volume for 2 hrs.…”

“…After 24 hr of co-culture, the incorporation of EPC-EXs into ECs was observed under a fluorescence microscope (EVOS; Thermo Fisher Scientific). The fluorescence intensity of ECs was analyzed using Image J as reported [28]. ECs were then used for various tests as described below.…”

Exosomes, the message transporters in vascular calcification

“…In fact, miR‐30 could be packed into exosomes and transport information among endothelial cells and mesenchymal stem cells . The expression of miR‐210 is also proven to be decreased in VC, and exosomal miR‐210 also ameliorated ROS production in the recipient endothelial cells . Such results indicated that exosomes could regulate the oxidative stress via modulating ROS production.…”

Exosomal cargo-loading and synthetic exosome-mimics as potential therapeutic tools

“…angiotensin II-induced apoptosis [62] . Anti-miR-9 oligonucleotides can be delivered by exosomes from MSCs to glioblastoma multiforme cells, where they sensitize the cells to antiglioblastoma multiforme drug treatment [63] .…”

“…Stress-treated cells miR-22 [56] , -in situ injection [56] from cells with miR-10a [57] modification Heat shock factor 1 [58] Genetic modified cells miR-143 [59,66] Tumor-targeting peptide [60] in situ injection [64] miR-let7a [60] iv injection [59,60] miR-146b [61] miR-210 [62] anti-miR-9 [63] miR-122 [64] miR-let7c [65] miR-155 and miR-125b2 [91] Exosomes loaded Electroporation BACE1 siRNA [67] Neuron-targeting peptide [67] iv injection [67,74] with exogenous cargo Doxorubicin (drug) [74] Tumor-targeting peptide [74] Lipofection RAD51 siRNA [68] --Sonication HER2 siRNA [69] --CaCl 2 miR-15a and anti-miR-15a [70] inhalation [70] (function not tested) Coincubation…”