Abstract: SA-PO0151
Uncovering the Role of Perilipin-2 in Successful Repair of Proximal Tubules Following Injury
Session Information
- AKI: Mechanisms - 3
November 08, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Biehl, Matthew J, Washington University in St Louis, St. Louis, Missouri, United States
- Li, Haikuo, Yale University, New Haven, Connecticut, United States
- Humphreys, Benjamin D., Washington University in St Louis, St. Louis, Missouri, United States
Background
Acute kidney injury (AKI) occurs frequently in hospitalized patients and the duration of injury or number of injury events contributes to the development of chronic kidney disease (CKD). The mechanisms governing repair of the proximal tubule (PT) remain poorly understood. We have previously utilized single-cell analysis to identify a unique subpopulation of injured PT cells in AKI induced by ischemia-reperfusion (IRI). One of the most highly upregulated genes identified codes for the lipid droplet-coating protein Plin2, which may suggest a crucial role for proper lipid droplet utilization in repair, perhaps through cellular autophagy.
Methods
We are currently utilizing both in vivo and in vitro models to knock out Plin2 to elucidate the mechanisms involved in PT repair. In vivo, we are using the well-established Pax8-rtTA; TetO-Cre model bred to Plin2-floxed mice to conditionally knock out Plin2 within the entire tubule. In vitro, we have adopted previously established protocols whereby we culture a PT-enriched population of epithelial cells from either cortical tissue of Plin2-floxed mice or donated human kidneys. Plin2 has then been successfully knocked out with either adenovirus-Cre or Plin2 siRNA, respectively.
Results
Preliminarily, following induction of Cre-recombinase with doxycycline we observed an obvious reduction of Plin2 expression in the PT by immunofluorescence (IF) 12 hours post IRI. We have also shown >90% reduction in Plin2 gene expression in vitro in both PT cell models following knock down with adenovirus-Cre or siRNA. Co-treatment of PT cells with the fatty acids oleic or palmitic acid (OA or PA) induced Plin2 expression at 6 hours post treatment by qPCR, and this induction is blunted following Plin2 knockdown. Finally, we have shown that lipid droplet formation is dependent upon Plin2 expression in PT cells in vitro, as knockdown of Plin2 results in an apparent reduction of BODIPY+ droplets in primary cells co-treated with OA.
Conclusion
These preliminary data set the framework to better understand the mechanism by which Plin2 is involved in repairing injured PT cells. Experiments are ongoing, and include investigation of cell autophagy, long-term recovery and multi-omics following injury in Plin2 knockout mice.
Funding
- NIDDK Support