Abstract: SA-PO0123
Unraveling the Metabolic Signature of Pkhd1-Deficient Renal Epithelial Cells
Session Information
- ADPKD and Cystic Kidney Disease - 3
October 24, 2026 | Location: Exhibit Hall A, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1201 Genetic Diseases of the Kidneys: Cystic (Monogenic)
Authors
- Cassina, Laura, IRCCS Ospedale San Raffaele, Milan, Lombardy, Italy
- Stefanoni, Davide, IRCCS Ospedale San Raffaele, Milan, Lombardy, Italy
- Distefano, Gianfranco, IRCCS Ospedale San Raffaele, Milan, Lombardy, Italy
- D'Anna, Melissa, IRCCS Ospedale San Raffaele, Milan, Lombardy, Italy
- D'Alessandro, Angelo, University of Colorado Anschutz Medical Campus, Aurora, Colorado, United States
- Boletta, Alessandra, IRCCS Ospedale San Raffaele, Milan, Lombardy, Italy
Background
Autosomal Recessive Polycystic Kidney Disease (ARPKD) is a rare genetic disease caused mainly by loss-of-function mutations in the PKHD1 gene, encoding fibrocystin/polyductin (FPC) protein. The molecular mechanisms underlying this form of PKD remain poorly understood. We contributed to show that Pkhd1-deficient renal epithelial cells display impaired mitochondrial respiration, which can be rescued by re-expression of a functional FPC fragment. These findings indicate that FPC is essential for mitochondrial homeostasis, and we are investigating whether its loss induces a metabolic shift, which may support abnormal cell proliferation.
Methods
Experiments are performed on pools of three single-cell clones of control and Pkhd1 KO mouse inner medullary collecting duct cell lines obtained by CRISPR/Cas9 technology. Oxygen consumption and extracellular acidification rates are assessed via Seahorse XF technology (Agilent Technologies), growth kinetics are profiled using high-throughput non-invasive Incucyte live-cell imaging and analysis system (Sartorius), functional metabolic dynamics are determined by unlabeled and labeled targeted metabolomics, using stable isotope tracers coupled to liquid chromatography–mass spectrometry.
Results
We assessed cell metabolism first via Seahorse technology, and we detected increased extracellular acidification coupled to reduced mitochondrial respiration in Pkhd1 KO cells. We performed LC-MS metabolomics and observed distinct clustering of metabolites between control and Pkhd1 KO cells by principal component analysis, with 68 of 347 metabolites significantly altered, including reduced TCA cycle intermediates and glutathione. Initial tracing data, showing decreased labelled glucose and increased labelled lactate, clearly suggest a glycolytic switch. These findings define a metabolic signature that warrants further investigation.
Conclusion
Our findings suggest that Pkhd1 loss triggers a metabolic rewiring in renal cells, primarily characterized by mitochondrial dysfunction and a shift toward glycolysis. Identifying these specific metabolic alterations is crucial for understanding ARPKD pathogenesis and may uncover novel therapeutic targets aimed at restoring metabolic homeostasis to limit cyst progression.
Funding
- Private Foundation Support