Eleonora Scaccia (Mannheim/ DE), Stefanie Uhlig (Mannheim/ DE), Prof. Francesca Pagano (Rom/ IT), Prof. Dr. Harald Klüter (Mannheim/ DE), Prof. Dr. Karen Bieback (Mannheim/ DE)
Background
Aberrant expression of microRNAs (miRNAs) appears associated with the progression of renal disease. Mesenchymal stromal cells (MSCs) exert a protective effect in damaged renal cells through the release of cytokines, growth factors and extracellular vesicles. We speculate that MSC-conditioned medium or EVs contained within it exert a protective effect, by modulating miRNA expression in injured renal cells and by this post-transcriptional gene regulation.
Methods
To mimic in vitro injury and MSC-mediated protection, conditionally- immortalized proximal tubule epithelial cells (ciPTECs) were treated with cisplatin for 1h and then treated for a further 23h with cisplatin plus adipose-derived Stromal Cells (ASC)-derived conditioned medium (CM-ASC). Viability, metabolic activity and migratory capacity in a scratch wound healing assay were then assessed. Further, mRNAs were prepared from ciPTECs treated with/without cisplatin and with/without MSC-CM and assessed by small-RNASeq.
Results
Cisplatin-treated ciPTECs showed reduced viability and metabolic activity, ameliorated by ASC-CM. Further, cisplatin significantly reduced the migration in the scratch wound assay, CM levelled it up even exceeding control levels. EVs failed to reproduce the results obtained by the CM. Small-RNASeq analysis identified differentially expressed miRNAs in all groups, particularly in the cisplatin-ASC-CM group compared to the ASC-CM group. Some of these miRNAs are known to regulate apoptosis and cellular fitness, while others have not been described in this context yet and are subject to further investigation now.
Conclusion
Our results demonstrate that MSC-CM per se exerts a protective effect reducing cisplatin cytotoxicity in renal proximal tubule cells. miRNA profiles are changed, suggesting that MSC-CM affects post-transcriptional gene regulation.
Offenlegung Interessenkonflikt:
No conflicts of Interest.