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

Abstract

Serum-derived extracellular vesicles (sEV) from healthy donors display in-vivo pro-angiogenic properties. To identify patients that may benefit from autologous sEV administration for pro-angiogenic purposes, sEV angiogenic capability has been evaluated in type 2 diabetic (T2DM) subjects (D), in obese individuals with (OD) and without (O) T2DM, and in subjects with ischemic disease (IC) (9 patients/ group). sEV display different angiogenic properties in such cluster of individuals. miRNomic profile and tGfβ content in sEV were evaluated. We found that miR-130a and TGFβ content correlates with sEV in-vitro and in-vivo angiogenic properties, particularly in T2DM patients. Ingenuity Pathway Analysis (IPA) identified a number of genes as among the most significant miR-130a interactors. Gain-of-function experiments recognized homeoboxA5 (HOXA5) as a miR-130a specific target. Finally, ROC curve analyses revealed that sEV ineffectiveness could be predicted (Likelihood Ratio+ (LH+) = 3.3 IC 95% from 2.6 to 3.9) by comparing miR-130a and TGFβ content 'in Series'. We demonstrate that sEV from high cardiovascular risk patients have different angiogenic properties and that miR-130a and TGFβ sEV content predicts 'true ineffective sEVs'. These results provide the rationale for the use of these assays to identify patients that may benefit from autologous sEV administration to boost the angiogenetic process. Cardiovascular complications are relevant causes of morbidity and mortality in patients with diabetes and obesity 1. Recent evidence suggests that extracellular vesicles (EVs) may act as mediators of many pathophysiological processes 2-4. EVs are small vesicles that are released from different cell types under normal and pathological conditions 5,6. Increased levels of circulating EVs have been associated with vascular impairment and hypercoagulability, particularly in patients with diabetes and acute coronary syndrome, suggesting that they play a role in driving cardiovascular diseases 7,8. Moreover, increased levels of circulating EVs, mainly from platelets and endothelial cells, has been proposed as a hallmark of cell dysfunction 9. It has been extensively reported that EVs act as biological active vectors, and participate in the exchange of information between circulating and resident cells, including endothelial cells 2,10. It has also been proposed that platelet-derived EVs play a role in the pathogenesis of atherosclerosis 11. EVs act as biological intermediaries, and mainly do so by delivering proteins, active lipids and extracellular RNAs 12. However, the most frequently studied EV-mediated biological processes rely on microRNA (miRNA) transfer. miRNAs are a class of small noncoding RNAs post-transcriptionally regulating gene expression 13. miR-NAs are present in serum/plasma and it has been proposed that their distinctive expression patterns serve as disease fingerprints in many clinical settings 14. Moreover, it has been shown that activated platelets can transfer

Mesenchymal Stem Cell-Derived Extracellular Vesicles Protect Human Corneal Endothelial Cells from Endoplasmic Reticulum Stress-Mediated Apoptosis

“…We previously analyzed the miRNA content of both MSC-EVs and SER-EVs [ 30 , 31 ]. In order to explore the mechanisms that account for the MSC-EV biological activity, we performed a Funrich analysis, combining the mostly expressed miRNAs together with the miRNAs predicted to target the main ER stress genes ATF4, CHOP, and XBP1.…”

Differential Therapeutic Effect of Extracellular Vesicles Derived by Bone Marrow and Adipose Mesenchymal Stem Cells on Wound Healing of Diabetic Ulcers and Correlation to Their Cargoes

“…This can possibly explain the beneficial effect of both EV types on endothelial cells. Moreover, several miRNAs expressed in both MSC-EVs could explain EV proangiogenic effect including miR-31-5p [ 44 ], miR-125a-5p [ 45 ], miR-126-3p [ 18 , 46 ]; miR-221-3p [ 20 ], miR-130a [ 47 ] and miR-132 [ 48 ]. In particular, our group already demonstrated the relevance of miR-126 for EV proangiogenic activity showing that EVs derived from ADSCs from obese patients have a reduced proangiogenic ability due to a reduced miR-126 content [ 49 ].…”

Extracellular Vesicles in Comorbidities Associated with Ischaemic Heart Disease: Focus on Sex, an Overlooked Factor

“…Since EV may be considered novel biomarkers of diabetes and other CVDs [ 20 , 45 , 46 ], discrete sex-dependent EV cargo differences, due to hormonal modulation, should be considered as they may help to explain the different risks of developing ischaemic conditions in men and women. In this regard, EV are certainly promising biomarkers of comorbidities for cardiac ischaemic conditions [ 23 , 24 ]; however, little attention has been paid to sex differences ( Table 1 ).…”

“…Although for a long time EV were considered only as double-layer phospholipid membrane vesicles and not biologically significant particles, they have emerged as biological entities able to influence the behaviour of target cells [ 19 ]. The effect of EV depends not only on their origin but also on the microenvironment containing the donor cells [ 20 , 21 , 22 ]. Due to their ability to carry their cargo, consisting of proteins, lipids, aminoacids, mRNAs, and miRNAs, circulating EV can be exploited as promising biomarkers for comorbidities of ischaemic conditions [ 23 , 24 ].…”