Abstract: FR-PO995
Urinary Single-Cell RNA Sequencing of Kidney Allograft Recipients Detects Podocyte Epithelial-to-Mesenchymal Transition in Kidney Transplant Recipients
Session Information
- Transplantation: Basic
October 25, 2024 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Transplantation
- 2101 Transplantation: Basic
Authors
- Muthukumar, Thangamani, Weill Cornell Medicine, New York, New York, United States
- Belkadi, Aziz, Weill Cornell Medicine- Qatar, Doha, Qatar
- Li, Carol Y., Weill Cornell Medicine, New York, New York, United States
- Thareja, Gaurav, Weill Cornell Medicine- Qatar, Doha, Qatar
- Dadhania, Darshana M., Weill Cornell Medicine, New York, New York, United States
- Suthanthiran, Manikkam, Weill Cornell Medicine, New York, New York, United States
Background
Podocytes undergo epithelial-to-mesenchymal transition (EMT) under states of cellular stress. We did urinary scRNA-Seq of kidney transplant recipients and detected the presence of podocyte EMT.
Methods
We obtained urine samples from 12 kidney transplant recipients (7 biopsy-matched urine samples [3 ABMR, 1 TCMR, and 3 no-rejection] and 5 stable patients without biopsy). We processed them fresh for scRNA-Seq on a 10x Chromium platform/Illumina sequencer. We used the Seurat package for computational analysis. We defined cell types based on the expression of established lineage marker genes.
Results
We generated 19404 high-quality single-cell transcriptomes from the urine of 12 kidney transplant recipients (Fig.1). Extracellular matrix (ECM) includes collagens, glycoproteins, and proteoglycans. We applied the single-cell ECM expression score (Kuppe et al. Nature 2021) to various kidney parenchymal cells. The podocyte score was highest among the kidney parenchymal cells and was similar to the myofibroblast score (Fig.2 left). To confirm podocytes' EMT, we ordered cells pseudo-temporally using scVelo (uses RNA velocity graph to draw descendants/ancestors coming from specified starting cells). Our analyses revealed a potential transition from podocytes (starting cells), but not other cells, to myofibroblasts (terminal states) (Fig.2 right).
Conclusion
By scRNA-Seq of urinary cells, we detected EMT in the podocytes of kidney transplant recipients. The role of the podocyte EMT in kidney allografts has not been well studied. Further characterization of podocyte EMT will help decipher the mechanisms underlying kidney allograft fibrosis.
Funding
- Other NIH Support