Abstract: SA-PO432
Human Mesenchymal Stem Cells Modulate High Glucose-Induced Inflammatory Responses of Renal Proximal Tubular Cell Monolayers and Their Cross-Talk with Macrophages
Session Information
- Development and Regenerative Medicine
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 500 Development, Stem Cells, and Regenerative Medicine
Authors
- Islam, Md Nahidul, National University of Ireland Galway, Galway, Ireland
- Griffin, Tomas P., University College Hospital Galway, Galway, Ireland
- McMorrow, Tara, University College Dublin, Dublin, Ireland
- Griffin, Matthew D., National University of Ireland, Galway, Galway, Ireland
Background
Renal proximal tubular epithelial cells (RPTEC) are dysfunctional in diabetic kidney disease (DKD). Mesenchymal stem cells (MSC) have been shown to modulate DKD pathogenesis. Having previously observed that soluble products of human MSC suppress high glucose (HG)-induced inflammatory responses of RPTEC/TERT1 stable monolayers, we aimed here to characterize the modulatory effect of MSC indirect co-culture on the transcriptional profile of RPTEC monolayers and to further explore the influence of MSC on RPTEC/Macrophage crosstalk.
Methods
Human RPTEC/TERT1 cells were cultured for 12 days to generate stable confluent monolayers. Normal medium or medium supplemented with 25mM D-Glucose (HG) or 25mM D-Mannitol (MAN) were applied for a further 5 days. Human bone marrow MSC were co-cultured 1:10 with RPTEC monolayers for the final 2 days in a trans-well system. RNA Sequencing, qRT-PCR and ELISA were performed on resulting samples. Conditioned Media from HG- and MAN-exposed RPTEC/MSC co-cultures were applied to monocyte-derived human macrophages under HG and MAN conditions.
Results
Bioinformatics analysis of RNA-sequencing data confirmed a predominant effect of HG on inflammation-related mediators and biological processes/KEGG pathways in RPTEC/TERT1 stable monolayers as well as the anti-inflammatory effect of MSC. KEGG pathway analysis showed HG-induced gene upregulation within the TNF-signalling, cytokine-cytokine receptor interaction and NOD-like receptor signalling. These gene expression signatures were modulated toward control expression by MSC co-culture. The HG-induced increase in RPTEC monolayer expression of transcripts for multiple cytokine (IL-1β, IL-6, IL-8, TNFα, MCP-1) and for NGAL as well as their counter-regulation by MSC were confirmed by qRT-PCR and ELISA. Conditioned medium from HG-exposed RPTEC/MSC transwell co-cultures attenuated secretion of inflammatory mediators (IL-8, TNFα, MCP-1) by macrophages compared to medium from HG-stimulated RPTEC alone.
Conclusion
Stable RPTEC monolayers demonstrate a delayed pro-inflammatory response to HG that is attenuated by close proximity to human MSC. In DKD, this MSC effect has potential to modulate hyperglycaemia-associated RPTEC/macrophage cross-talk - a key pathogenic mechanism of chronic interstitial inflammation.