Abstract: SA-PO302

Adult Renal Stem/Progenitor Cells Can Revert LPS-Induced Endothelial-to-Mesenchymal Transition of Endothelial Cells

Session Information

Category: Cell Biology

  • 204 Extracellular Matrix Biology, Fibrosis, Cell Adhesion

Authors

  • Sallustio, Fabio, University of Bari, Bari, Italy
  • Lucarelli, G., University of Bari, Bari, Italy
  • Battaglia, Michele, University of Bari, Bari, Italy
  • Castellano, Giuseppe, University of Bari, Bari, Italy
  • Gesualdo, Loreto, University of Bari, Bari, Italy
  • Stasi, Alessandra, University of Bari, Bari, Italy
  • Curci, Claudia, University of Bari, Bari, Italy
  • Franzin, Rossana, University of Bari, Bari, Italy
  • Divella, Chiara, University of Bari, Bari, Italy
  • Laghetti, Paola, University of Bari, Bari, Italy
  • De Palma, Giuseppe, University of Bari, Bari, Italy
  • Picerno, Angela, University of Bari, Bari, Italy
  • Rutigliano, Monica, University of Bari, Bari, Italy
Background

Acute Kidney Injury (AKI) is the major complication encountered in sepsis. Lipopolysaccharides (LPS) are frequently involved in the pathogenesis of AKI, that is mainly characterized by endothelial cell (EC) dysfunction. EC acquire a myo-fibroblast phenotype, by endothelial-to-mesenchymal transition (EndMT), contributing to the renal fibrosis. Resident adult renal stem/progenitor cells (ARPCs) enhance tubular regenerative mechanism during AKI, but little is known about their effects on endothelial compartment. The aim of this study is to investigate the effects of ARPCs on endothelial dysfunction.

Methods

Endothelial cells were stimulated in vitro with LPS for 48h and co-cultured with ARPCs for 24h. MTT cell viability assay was used to analyze the EC proliferation rate following LPS stimulation and in co-culture with ARPCs. FACs analysis was used to study the expression of myofibroblast markers. Gene expression profiles of ARPCs and EC were generated using Agilent Microarrays.

Results

We observed a significant increase of EC proliferation after stimulation with LPS. ARPCs in co-culture with EC normalized their proliferation rate and decrease the cell growth rate, even in presence of LPS. Moreover, LPS induced a significant decrease of EC markers, CD31 and VE-cadherin and a significant increase of EC dysfunction markers, Collagen I and Vimentin. ARPCs in co-culture with EC abrogated the LPS-induced EndMT by restoring the high expression of CD31 (95% vs 66%) and VE-cadherin (96% vs 31%) and limiting Collagen I (18% vs 73%) and Vimentin (35 % vs 50.86%) expression.
Microarray analysis showed that LPS induced the upregulation of 305 genes and the down regulation of 694 genes in ARPCs (Q value < 0,05 and Fold change >2). Gene Set Enrichement Analysis and pathway analysis identified 27 genes specifically involved in prevention and recovery from infections caused by external agents.

Conclusion

Our data demonstrate that ARPCs could preserve EC phenotype by regulating LPS-induced EndMT. Interestingly, LPS induces the expression of a specific gene set in ARPCs able to prevent EC dysfunction.