Table 2.
EV-bound miRNAs in the regulation of cardiovascular neovascularisation.
| EV miRNA | Model | Effect | EV source | EV isolation method | References |
|---|---|---|---|---|---|
| miR-15b miR-17 miR-20a miR-103 miR-199a miR-210 miR-292 |
Rat cardiac endothelial cells Male Sprague-Dawley rats- MI induction by LAD coronary artery ligation |
Increased EC tube formation post-hypoxic EV treatment in-vitro. Improved cardiac function and reduced fibrosis post-intramyocardial EV injection at 3 border zones. EV microRNA expression profiling by qPCR miRNA array revealed several upregulated miRNAs under hypoxic conditions | Normoxic/ hypoxic CPCs | Differential centrifugation ultracentrifugation |
Gray et al., 2015 |
| miR-16 miR-17-92 miR-19b miR-20a miR-34 miR-126-3p miR-130a-3p miR-210-3p miR-294 |
Murine cardiac ECs. C57BL6/J mice- MI induction by 30-min coronary occlusion followed by reperfusion | Increased EC tube formation, migration and antiapoptotic properties post-EV treatment in-vitro. Increased capillary density & reduced LV remodelling and hypertrophy post-intramyocardial injection of EVs at 5 sites at the border between infarcted and non-infarcted myocardium 48h post-reperfusion. EV microRNA expression profiling was performed by miRNA array and revealed several upregulated miRNAs. | iPSCs | Differential centrifugation ultracentrifugation |
Adamiak et al., 2018 |
| miR-17 miR-19a miR-19b miR-20a miR-30c miR-126 |
HUVECs | Increased EC tube formation, proliferation and migration post-exosome treatment in-vitro. EV microRNA expression profiling was performed by qPCR miRNA array and revealed increased levels of several miRNAs in these exosomes | Glucose starved H9C2 cardiomyocytes | Differential centrifugation ultracentrifugation |
Garcia et al., 2015 |
| miR-20b miR-27b miR-29b miR-42a miR-100 miR-125b miR-143 miR-195 miR-291b miR-497 |
CMVECs Aortic rings from Male Sprague-Dawley rats Male Sprague-Dawley rats- HLI induction by femoral artery ligation |
Increased EC tube formation, proliferation and migration post-exosome treatment in-vitro. Increased EC sprouting post-exosome treatment ex-vivo. Increased capillary density and blood perfusion post-induced vascular progenitor cell (iVPC) exosome injection as compared with rat aortic endothelial cell (RAEC) exosomes and controls. EV microRNA expression profiling was performed by qPCR miRNA array and revealed increased levels of several miRNAs in these exosomes | iVPCs RAECs |
Ultracentrifugation Ultrafiltration Size-exclusion chromatography |
Johnson et al., 2019 |
| miR-21-5p | HUVECs Female CD-1 mice- MI induction by LAD coronary artery ligation |
Improved EC tube formation in-vitro post-treatment with EVs from patients with normal angiography results (NEXO) compared to patients with heart failure (FEXO). Increased capillary density, infarcted wall thickness and decreased infarct size post-NEXO EV treatment compared to FEXO EVs and PBS controls post-intramyocardial injection. Decreased miR-21-5p expression in FEXO compared to NEXO group identified by EV microRNA expression profiling by qPCR miRNA array. Silencing of miR-21-5p in NEXO group decreased tube-formation in-vitro, while upregulation of miR-21-5p in FEXO group promoted EC tube formation. | Explant-derived cardiac stromal cells from FEXO or NEXO patients | Ultracentrifugation | Qiao et al., 2019 |
| miR-21 miR-27a miR-29a miR-126 miR-130α miR-191 miR-210 miR-296-3p |
HMVECs Human macrovascular endothelial cells (HMAVECs) SCID mice |
Increased EC tube formation and proliferation post-treatment with EVs from obese individuals with type 2 diabetes (OD) compared with EVs from healthy individuals (H), patients with type 2 diabetes (D), obese individuals without T2DM (O), and patients with ischaemic disease (IC). EVs from patients of the above groups were divided as “effective” and “ineffective” depending on their effect on angiogenesis in vitro. Matrigel plug in vivo assays using EVs defined as “effective” in the in vitro assays, showed that these EVs resulted in an increased number of vessels in mice Matrigel plugs. EV microRNA expression profiling of “effective” and “ineffective” EVs of healthy individuals was performed by qPCR miRNA array and identified several angiogenic miRNAs | Human serum from H, D, OD, D, O, IC individuals | Ultracentrifugation | Cavallari et al., 2020 |
| miR-31 | HUVECs Aortic rings from male C57BL/6J mice |
Increased EC tube formation and migration post-EV treatment in-vitro. Increased EC sprouting post-EV treatment ex vivo. EV microRNA expression profiling was performed by qPCR miRNA array and revealed increased levels of miR-31. EVs from cells transfected with anti-miR-31 decreased EC tube formation and migration in-vitro, whereas pre-miRNA transfection increased tube formation and migration. | Adipose-Derived Stem Cells (ADSCs) | Differential centrifugation Ultracentrifugation |
Kang et al., 2016 |
| miR-92a-3p | HCAEC | EV microRNA expression profiling was performed by qPCR miRNA array and revealed increased miR-92a-3p levels in oxidised low-density lipoprotein (oxLDL) and interleukin-6 (IL-6) stimulated HCAEC. Knock-down of this miRNA in HCAEC EVs decreased their ability to cause HCAEC migration, proliferation, and tube formation in-vitro. | oxLDL and IL-6 stimulated HCAEC | Differential centrifugation Ultracentrifugation |
Liu et al., 2019 |
| miR-125a-5p | HUVEC | Increased EC tube formation post-EV treatment in-vitro. EV microRNA expression profiling was performed by qPCR miRNA array. Overexpression of miR-125a in HUVECs, increased tube formation in-vitro. Matrigel plug in vivo assay results showed that the number of vascular structures was significantly higher in mice subcutaneously injected with HUVECs mixed with EVs than HUVECs alone | MSCs | Differential centrifugation Ultrafiltration Ultracentrifugation |
Liang et al., 2016 |
| miR-126-3p | HUVECs Immunocompromised BalbC mice- HLI induction by femoral artery ligation |
Increased EC tube formation post-EV treatment in-vitro. EV microRNA expression profiling was performed by qPCR miRNA array. EVs from cells transfected with anti-miR-126-3p decreased HUVEC tube formation in-vitro. Improved blood flow and capillary density post-intramuscular injections of EVs at 4 different sites of the ischaemic limb. | CD34+ SCs | Ultracentrifugation Density gradient centrifugation |
Mathiyalagan et al., 2017 |
| miR-126 miR-296 |
SCID mice- HLI induction by left femoral artery and the distal portion of saphenous artery ligation | Increased capillary density and blood perfusion post-MV injection. Presence of miR-296/-126 in the EVs was confirmed by qPCR | EPCs | Ultracentrifugation | Ranghino et al., 2012 |
| miR-132 miR-146a-3p miR-181 miR-210 |
HUVECs Male Wistar rats- MI induction by LAD coronary artery ligation |
Increased EC tube formation ability post-EV treatment in-vitro. EV microRNA expression profiling was performed by qPCR miRNA array. HUVECs transfected with a miR-132 mimic had increased tube formation. Opposite results were observed post-transfection with si-miR-132. In-vivo experiments revealed increased blood vessel density post-intramyocardial injection of CPC EV at 3 border zones. | hCPCs | ExoQuick™ precipitation solution (System Biosciences; SBI) Ultracentrifugation column precipitation using Exo-spin™ kits (Cell Guidance Systems, Cambridge, UK) |
Barile et al., 2014 |
| miR-143 miR-222 |
MCECs HUVECs Rat aortic rings Fertilised chicken eggs Female C57BL/6 mice- MI induction by LAD coronary artery ligation |
EVs from cells in ischaemic solutions increased EC tube formation, proliferation and protection against oxidative-induced lesion in-vitro and increased EC sprouting ex-vivo. Intramyocardial injection of hypoxic EVs increased capillary density, the number of CD31+ cells in the infarcted region and blood perfusion. EV microRNA expression profiling by qPCR miRNA array revealed increased miR-222/-143 expression in hypoxic EVs | H9c2 myocardial cells & primary rat cardiomyocytes from Wistar rat foetus cultured in appropriate media or ischaemia-mimetic solutions | Differential centrifugation Density gradient centrifugation |
Ribeiro-Rodrigues et al., 2017 |
| miR-210 | HUVEC Male C57BL/6 mice- MI induction by LAD coronary artery ligation |
Increased EC tube formation & decreased apoptosis post-hypoxic EV treatment in-vitro. EV microRNA expression profiling by qPCR miRNA array revealed increased miR-210 expression in hypoxic EVs. Transfection of HUVECs with miR-210 mimic yielded similar results to hypoxic EV treatment. Increased capillary and arteriole density and CM survival post-intramyocardial injection of hypoxic EVs at five sites around the border zone of infarcted hearts. | Normoxic/ hypoxic MSCs | Differential centrifugation Ultracentrifugation |
Zhu et al., 2018 |
| miR-214 | HMVECs | Increased EC tube formation and migration post-EV treatment in-vitro. EV microRNA expression profiling was performed by qPCR miRNA array and showed increased miR-214 expression. EVs from cells transfected with anti-miR-214 decreased EC tube formation and migration in-vitro, whereas pre-miRNA transfection did not affect tube formation and migration. | HMVECs | Differential centrifugation Density gradient centrifugation |
Balkom et al., 2013 |
| miR-423-5p | HUVECs | Increased EC tube formation, proliferation and migration post-exosome treatment in-vitro. EV microRNA expression profiling by small RNA sequencing and qPCR miRNA array revealed increased levels of miR-423-5p in these exosomes. Overexpression of miR-423-5p in HUVECs increased their tube formation ability, proliferation and migration. | hADSCs | Differential centrifugation Ultracentrifugation |
Xu et al., 2019 |
| miR-939-5p | MCECs HUVECs C57BL/6 mice- MI by LAD coronary artery ligation & HLI by left femoral artery ligation |
Increased EC tube formation, proliferation and migration post-ischaemic exosome treatment in-vitro. Increased capillary density and reduced scar size post-ischaemic exosome injection. EV microRNA expression profiling by qPCR miRNA array revealed decreased miR-939-5p expression in ischaemic exosomes. HUVEC transfection with miR-939-5p mimic decreased EC tube formation and migration in-vitro, whereas transfection with miRNA inhibitor increased tube formation and migration. | Coronary blood from patients with myocardial ischaemia and control group | Differential centrifugation Ultracentrifugation |
Li et al., 2018 |
| miR-1246 | HUVECs | Incubation of HUVECs with DLD-1 cancer cell derived EVs significantly increased tube formation and migration. Mir-1246 was enriched in DLD-1-EVs. Overexpression of miR-1246 in HUVECs significantly increased their tube formation ability and migration. | DLD-1 cancer cells | Ultracentrifugation | Yamada et al., 2014 |
| miR-4306 | HCAECs | Decreased EC tube formation, migration and proliferation post-treatment with EVs isolated from the PCI group compared with those from the control group. Human monocyte-derived macrophage (HMDM) treatment with ox-LDL significantly increased their miR-4306 levels. Transfection of HCAECs with miR-4306 mimic significantly inhibited their tube formation ability and slightly suppressed HCAEC proliferation and migration. | HMDM from patients who had PCI performed within 12 hours (PCI group) and patients with chest pain syndrome with normal coronary artery findings post coronary angiography | Differential centrifugation Ultracentrifugation |
Yang et al., 2019 |
| let-7b-5p | HUVECs CD1 male mice-HLI induction by left femoral artery ligation |
Increased EC tube formation, proliferation and reduced EC apoptosis post-exosome treatment in-vitro. EV microRNA expression profiling was performed by qPCR miRNA array and revealed increased levels of let-7b-5p. Exosomes restored the angiogenic capacity of DICER knock-out-ECs, but the reduction of exosomal let-7b-5p in exosomes failed to induce tube formation in recipient DICER-KD ECs in-vitro. Increased capillary density and blood flow recovery and reduced necrosis post-exosome injection in-vivo. | Human pericardial fluid (PF) exosomes | ExoQuick kit (System Biosciences) | Beltrami et al., 2017 |