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Published on: November 8, 2024
MicroRNA Cargo in Wharton's Jelly MSC Small Extracellular Vesicles: Key Functionality to In Vitro Prevention and
Vera Tscherrig1,2,3, Sophie Cottagnoud1,3, Valérie Haesler1,3
1Department of Obstetrics and Feto-maternal Medicine, University Women's Hospital, Inselspital, Bern University Hospital, Bern, Switzerland.
Insights
Mesenchymal stromal cell-derived small extracellular vesicles (MSC-sEV) show therapeutic potential for preterm birth-related white matter injury. Their microRNAs are key to protecting against neurological damage by regulating gene expression and promoting cell survival.
Area of Science:
- Neuroscience
- Regenerative Medicine
- Molecular Biology
Background:
- Preterm birth causes white matter injury (WMI), leading to long-term neurological deficits.
- Mesenchymal stromal cell-derived small extracellular vesicles (MSC-sEV) are a potential therapy for perinatal brain injury.
- MicroRNAs (miRNAs) within MSC-sEV are hypothesized to mediate therapeutic effects.
Purpose of the Study:
- To investigate the role of miRNAs in MSC-sEV for treating WMI.
- To identify specific miRNAs in MSC-sEV and their target genes.
- To evaluate the impact of MSC-sEV on oligodendroglial maturation and neuronal apoptosis.
Main Methods:
- Isolation and purification of small extracellular vesicles (sEV) from umbilical cord Wharton's jelly MSC.
- Quantification of miRNA content and validation of target gene silencing (TP53, TAOK1) using luciferase assays.
- In vitro assessment of sEV effects on oligodendroglial maturation and neuronal apoptosis using an oxygen-glucose deprivation/reoxygenation (OGD/R) model, with DROSHA knockdown to assess miRNA processing impact.
Main Results:
- WJ-MSC-sEV contained specific miRNAs (hsa-miR-22-3p, hsa-miR-21-5p, hsa-miR-27b-3p, let-7 family) implicated in WMI.
- sEV miRNAs demonstrated inhibitory effects on TP53 and TAOK1 gene expression.
- sEV treatment promoted oligodendrocyte maturation and reduced neuronal apoptosis in an OGD/R model.
- Disruption of miRNA processing via DROSHA knockdown abolished the beneficial effects of sEVs.
Conclusions:
- MicroRNAs play a crucial role in the therapeutic efficacy of WJ-MSC-sEV for preterm WMI.
- MSC-sEVs exert neuroprotective effects by modulating specific miRNA pathways.
- This study highlights the potential of MSC-sEVs as a cell-free therapy for perinatal neurological injuries.
Abstract:
Preterm birth is the leading cause of childhood morbidity and mortality and can result in white matter injury (WMI), leading to long-term neurological disabilities with global health burden. Mesenchymal stromal cell-derived small extracellular vesicles (MSC-sEV) are a promising therapeutic agent for treating perinatal neurological injury. They carry microRNAs (miRNAs) predicted to be involved in the onset of premature WMI. We hypothesize that miRNAs have a key function in the beneficial effects of MSC-sEV. We isolated MSC from umbilical cord tissue, the Wharton's jelly (WJ), and purified small extracellular vesicles (sEV) from WJ-MSC culture supernatant by ultracentrifugation and size exclusion chromatography. The miRNA content was quantified by real-time polymerase chain reaction. A luciferase gene assay validated silencing of TP53 and TAOK1, which we previously identified as predicted target genes of MSC-sEV miRNAs by Next Generation Sequencing and pathway enrichment analysis. The impact of sEV miRNAs on oligodendroglial maturation and neuronal apoptosis was evaluated using an in vitro oxygen-glucose deprivation model (OGD/R) by knocking-down DROSHA in WJ-MSC, which initiates miRNA processing. WJ-MSC-sEV contained miRNAs involved in WMI, namely hsa-miR-22-3p, hsa-miR-21-5p, hsa-miR-27b-3p, and the hsa-let-7 family. The luciferase assay strongly indicated an inhibitory effect of sEV miRNAs on the gene expression of TP53 and TAOK1. Small EV initiated oligodendrocyte maturation and reduced OGD/R-mediated neuronal apoptosis. Knocking-down DROSHA in WJ-MSC reduced the expression of sEV miRNAs and led to the loss of their beneficial effects. Our in vitro study strongly indicates the key function of miRNAs in the therapeutic potential of WJ-MSC-sEV in premature WMI.
