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Isolating, Sequencing and Analyzing Extracellular MicroRNAs from Human Mesenchymal Stem Cells
Published on: March 8, 2019
Exosome-Encapsulated microRNA-127-3p Released from Bone Marrow-Derived Mesenchymal Stem Cells Alleviates
Jisheng Dong1, Li Li1, Xing Fang1
1Department of Orthopedics, The Second People's Hospital of Hefei, The Affiliated Hefei Hospital of Anhui Medical University, Hefei, Anhui, 230011, People's Republic of China.
Objective:
Exosome-encapsulated microRNAs (miRNAs) are being considered as either diagnostic or predictive markers in different types of diseases. Here, we discussed the effects of exosome-encapsulated miR-127-3p from bone marrow-derived mesenchymal stem cells (BM-MSCs) on osteoarthritis (OA).
Methods:
BM-MSCs and primary chondrocytes were isolated from Sprague Dawley rats. IL-1β was utilized to treat chondrocytes to mimic an OA in vitro model, and exosomes extracted from BM-MSCs were utilized to treat chondrocytes so as to verify their protective effects on OA. Through online website prediction and experiments confirmation, we found the most significantly enriched miRNA in exosomes and elucidated the effect of this miRNA on the therapeutic effect of exosomes by interfering with its expression. Also, the genes targeted by the miRNA and the involved pathway were also found through bioinformatics analysis and experimental research, thereby probing into the protective mechanism of exosomes on chondrocytes.
Results:
Exosomes derived from BM-MSCs restricted the IL-1β-induced chondrocytes damage. miR-127-3p was found to be enriched in exosomes, and the protective effect of exosomes was reversed by miR-127-3p inhibition. miR-127-3p targeted CDH11, and overexpressed CDH11 in chondrocytes weakened the therapeutic effect of exosomes. IL-1β treatment resulted in the activation of the Wnt/β-catenin pathway in chondrocytes. Exosomes treatment could inhibit the activation of this pathway, and overexpressed CDH11 reversed the inhibitory effect of exosomes on this pathway.
Conclusion:
This study suggests that exosomal miR-127-3p derived from BM-MSCs inhibits CDH11 in chondrocytes, thereby blocking the Wnt/β-catenin pathway activation and relieving chondrocyte damage in OA.
Insights
Exosomes from bone marrow-derived mesenchymal stem cells (BM-MSCs) protect against osteoarthritis (OA) by delivering miR-127-3p. This microRNA inhibits CDH11, blocking the Wnt/β-catenin pathway and reducing chondrocyte damage in OA.
Area of Science:
- Biomedical research
- Stem cell biology
- Molecular medicine
Background:
- Exosomes carry microRNAs (miRNAs) with potential as disease biomarkers.
- Osteoarthritis (OA) involves chondrocyte damage and inflammation.
- Bone marrow-derived mesenchymal stem cells (BM-MSCs) show therapeutic potential.
Purpose of the Study:
- To investigate the role of exosomal miR-127-3p from BM-MSCs in osteoarthritis (OA).
- To elucidate the protective mechanism of these exosomes on chondrocytes.
- To identify the molecular targets and pathways involved.
Main Methods:
- Isolation of BM-MSCs and primary chondrocytes from Sprague Dawley rats.
- In vitro OA model using IL-1β-treated chondrocytes.
- Exosome isolation and treatment, miRNA expression analysis, and bioinformatics prediction.
- Gene targeting and pathway analysis (Wnt/β-catenin).
Main Results:
- BM-MSC-derived exosomes protected chondrocytes from IL-1β-induced damage.
- miR-127-3p was enriched in exosomes and mediated their protective effect.
- miR-127-3p targeted CDH11, and its inhibition reversed exosome efficacy.
- Exosomes inhibited the Wnt/β-catenin pathway, an effect reversed by CDH11 overexpression.
Conclusions:
- Exosomal miR-127-3p from BM-MSCs protects against OA by targeting CDH11.
- This mechanism involves the inhibition of the Wnt/β-catenin pathway.
- BM-MSC exosomes represent a potential therapeutic strategy for OA.
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