Table 2.
Details of the experimental design of studies evaluating EV effector molecules. Study design, EV characteristic, target and main study findings are outlined
| CKD model |
Study | Cell | AD | CM | EV size | EV markers | Animal model | Effector | Target | Main findings |
|---|---|---|---|---|---|---|---|---|---|---|
| UUO | Wang [38] | MSCs (r): | IV | Column | n.a. | MSC-EV markers | Fisher 344 rat (UUO), In vitro (h TECs, TGF-β1) | miR-294, miR-133, (mimic/ inhibitor) | TGF-β1 | Old rat miRNA-depleted EVs failed to improve kidney fibrosis. Injection of mimic miRNAs reversed those effects. Also, overexpression of miRNAs mitigated TGF-β1-mediated EMT in TECs.. |
| UUO | Chen [10] | GDNF--MSCs (h) | IV | UC | 30-150 nm | CD9, CD63, CD81 | Nude mice* (UUO), In vitro (human EC) | GDNF (GDNF-EVs) | SIRT1/eNOS | GDNF-MSC-EVs were more effective in reducing fibrosis than GFP-MSC-EVs. However, GDNF-MSC-EVs did not improve renal function. |
| UUO | Ji [28] | MSCs (h) | IV | UC | 30-180 nm | CD9, CD63, Alix | SD rat (UUO), In vitro (rat TECs, mechanical stress) | CK1δ, β-TRCP (KO-EVs: shCK1δ- and shβ-TRCP-EVs) | Yes-associated protein (YAP) | EVs overexpressed CK1δ and β-TRCP protein. Administration of EVs increased CK1δ, β-TRCP, and decreased expression of YAP in kidney tissue. CK1δ, and β-TRCP knockdown decreased anti-fibrosis effectiveness of EVs. |
| UUO | Shi [37] | MSC (r)-silenced | IV | UC | 50-600 nm | CD9, CD63, HSP70 | SD rat (UUO), In vitro (human TECs, TGF-β1) | MFG-E8 (MFGE8–silenced-EVs) | RhoA/ROCK pathway | Protective effects of EVs on kidney histology, fibrosis, and inflammation were abolished by the inhibition of MFG-E8 in EVs. |
| UUO | Wang [41] | miR-29-Satellite cells (m) | IM | UC | 87-93 nm | TSG101 | C57BL/6J mouse (UUO) | miR-29 (miR-29-EVs) | TGF-β3 | Injection of miR29-EVs attenuated renal histology and fibrosis. |
| UUO | Zhang [43] | miR-26a-EKC (h) | IM | UC | 50-300 nm | TSG101 | C57BL/6J mouse (UUO) | miR-26a (miR-26a-EVs) | CTGF, TGF-β1 | Injection of miR-26-EVs attenuated renal fibrosis by limiting CTGF. |
| UUO | Wang [40] | let-7C-MSCs (h) | - | PC | n.a. | ? | In vitro (mouse TECs, TGF-β) | let-7c | TGF-β1 | let7c-EVs inhibited TGF-β1 in vitro |
| HT | Cambier [17] | CPCs (h) | IV | MF | n.a. | * | C57BL/6J mouse (Ang II) | Y-RNA | IL10 | YF1-EV-RNA improved kidney function, and diminished renal inflammation and fibrosis. |
| HT | Zou [47] | STC-like cells (p) | IR | UC | 20-310 nm | CD9, CD29, CD81 |
129-S1 mouse (RAS), In vitro (human TECs, AMA) |
mitochondia, mitochondial DNAs | - | STC-like cells-EV-mitochondria remained functional and acquired TEC function. |
| HT | Eirin [23] | MSCs (p) adipose tissue (IL-10 KD) | IR | UC | 30-400 nm | CD9, CD40, CD81 | Domestic pig (obese diet + RAS) | IL10 mRNA ( KO EVs) | - | Compared with MSC-EVs, the protective effects on kidney morpology, renal function, and macrophage phenotype were blunted for KO-EVs. |
| HT | Zhao [45] | MSCs (p), adipose tissue | IR | UC | 100-200 nm | CD9, CD29, CD69 | Domestic pig (obese diet + RAS) | miR-532-5p | Growth factors? | microRNA-532-5p expression was upregulated in stenotic kidneys, possibly by its delivery by EVs. |
| DN, T1D | Duan [20] | SCs (h) urine | IV | PC | 30–120 nm | CD63, TSG101, HSP90B1 calnexin - |
SD rat (Stz), In vitro (human podocytes, glu) |
miR-16-5p (miR-16‐5p-EVs and KO EVs) | VEGF | Compared with uSCs‐EVs, the hyperplasia of mesangial matrix and kidney function were more alleviated for miR-16-5p-EVs. |
| DN, T1D | Zhong [46] | MSCs (h), umbilical cord | IV | PC | 30-500 nm | MSC-EV markers |
Balb/C mouse, (Stz) with hyperurycemia In vitro (human PTCs) |
miR-451a | P15, P19 | Expression of miR was enriched in EVs compared with MSCs. Injection of EV-miR-451a (agomir) ameliorated tubular damage, and reduced EMT by inhibiting CKIs. |
| DN, T2D | Duan [21] | MSCs (m), adipose tissue | IV | UC | 30-150 nm | CD9, CD63, CD81, TSG101 | C57BL/KsJ db/db mouse, In vitro (mouse podocyte, glu) | miR-26a-5p (miR-26a-5p-EVs, KO EVs, miR antagomir) | TLR4 | Administration of EVs induced miR-26a-5p and decreased TLR4 expression in kidney tissue. KO of EVs failed to induce any improvement in kidney function or renal histology. Delivery of miR-26a -5p-EVs to podocythes reduced apoptosis. |
| DN, T2D | Jin [30] | MSCs (m), adipose tissue | - | Immuno-P | 60-500 nm | CD9, CD63, CD81 | In vitro (mouse podocyte, glu) | miR-486 (KO EVs) | Smad1 | miR-486 inhibition reduced the protective role of EVs in high glucose-induced podocyte damage. |
| DN, T2D | Jin [31] | MSCs (m), adipose tissue, miR-215-5p-KO | - | Immun-P | <100 nm | CD9, CD63, CD81 | In vitro (mice podocyte, glu) | miR-215-5p (KO EVs) | ZEB2 | KO EVs failed to modulate glucose-induced podocyte migration in vitro, while transfection with miR-215-5p mimics in podocytes reversed the effect. miR-215-5p (mimic) blocked HG-induced ZEB2 expression in vitro. |
| (Other)GN | Cantaluppi [18] | EPCs (h), blood | IV | UC | 60-130 nm | CD55, CD59 | Wistar rat (anti-Thy1.1 Ab), In vitro (rat mesangial cells, anti-Thy1.1 Ab) |
Factor H, CD55 and CD59 mRNAs, miR-126, miR-296 |
C5b-9 | EVs reduced cell damage and death in the presence of rat or human sera. These effects were blunted when EVs were treated with RNAse. |
| (Other) Alport | Sedrakyan [36] | AFSC (m) | IC | UC | 50-500 nm | CD9, CD63 , CD24 | Alport-TektdT mouse, In vitro (mice GEC) | miRNAs, VEGFR1, sVEGR1 (KO EVs) | VEGF | KO EVs failed to modulate kidney VEGF expression. |
Cell of EV origin: AFSC amniotic fluid stem cell, CPC cardiac progenitor cell, EKC embryonic kidney cell, EPC endothelial progenitor cell, MSC mesenchymal stem cell, SC satellite cell, STC STC-like cell, uSC urine stem cell. AD EV administration, CM EV concentration method, UC ultracentrifugation, PC precipitation, MF microfiltration, Immuno-P immuno-precypitation. Methods of EV measurement: DLS dynamic light scattering, TEM transmission Electron Microscopy, NTA nanoparticle tracking analysis, TRPS tunable resistive pulse sensing. EV markers: CD cluster of Differentiation, HLA human leukocyte antigen, TSG101 tumor Susceptibility 101. CKD model: HT hypertension, DM diabetes, RAS renal artery steatosis, UUO unilateral ureteral obstruction, Nx nephrectomy, AMA Antimycin-A, CyA cyclosporine A, PTC peritubular capillary, TEC tubular epithelial cell, TGF-β1 transforming growth factor β. Target NOS endothelial nitric synthase, CTGF Connective tissue growth factor, IL10 Interleukin 10, Smad 1 SMAD Family Member 1, TLR4 Toll-like receptor 4, EMT epithelial to mesenchymal transition