Abstract: TH-PO984
Proximal tubular cell isolation during human acute kidney injury
Session Information
- Glomerular Diseases: Minimal Change Disease, FSGS, IgAN
November 07, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- De Seigneux, Sophie M., Geneva University Hospital, Geneva, Switzerland
- Legouis, David, Geneva University Hospital, Geneva, Switzerland
- Galichon, Pierre, Sorbonne Universite, Paris, France
Background
There is an urgent need to discover and validate the pathophysiological pathways involved in human acute kidney injury (AKI). Proximal tubular cells (PTCs) are the cells primarily injured during AKI and their maladaptive repair contribute to AKI to chronic kidney disease (CKD) transition. It is very important to develop non-invasive tools that will help to improve our understanding of the signaling pathways contributing to the maladaptive repair.
Methods
We collected urine from patients hospitalized in the ICU of University Hospital of Geneva, presenting or not AKI. We here propose a novel method to isolate specifically PTCs from human urine using a combination of Magnetic-Activated Cell Sorting (MACS) sorting and Fluorescent-Activated Cell Sorting (FACS) . We have characterized the sorted cells, and show how these cells can be used to determine pathophysiological pathways in human AKI.
Results
Isolated PTCs are viable, can be cultured for at least three passages (a) and retain the expression of several proximal tubule specific markers at the level protein (b) and of mRNA (c). We report that our technique is more sensitive than routine microscopic approach used to identify the presence of tubular cells in the urine during clinical AKI (d). There is a clear correlation between isolated PTC cell numbers and the severity of AKI as defined by KDIGO staging guidelines (e). RNA sequencing and pathway analysis confirmed alterations in PTCs cell cycle in AKI patients, which correlated to the severity of AKI (f).
Conclusion
We show that isolation of PT cells from urine is possible during human AKI episode. These cells can be used to study the pathways involved in AKI and we could confirm alterations of the cell cycle in PTCs collected from AKI patients. Thus, we believe that PTCs isolated from human urine will allow the identification and characterization of novel pathways in AKI and the AKI to CKD transition.
Funding
- Private Foundation Support