Abstract: FR-PO0638
Pkd1 Deletion in Renal Fibroblasts Drives Spontaneous Differentiation to Migratory, Matrix-Producing Myofibroblasts with Altered Metabolism
Session Information
- Cystic Kidney Diseases: Basic and Translational Research
November 07, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1201 Genetic Diseases of the Kidneys: Monogenic Kidney Diseases
Authors
- Remadevi, Viji, The University of Kansas Medical Center, Kansas City, Kansas, United States
- Ward, Christopher J., The University of Kansas Medical Center, Kansas City, Kansas, United States
- Patel, Prisha S, University of Tennesse Health Science Center, Memphis, Tennessee, United States
- Bajwa, Amandeep, University of Tennesse Health Science Center, Memphis, Tennessee, United States
- Bohovyk, Ruslan, University of South Florida, Tampa, Florida, United States
- Levchenko, Vladislav, University of South Florida, Tampa, Florida, United States
- Staruschenko, Alexander, University of South Florida, Tampa, Florida, United States
- Jamadar, Abeda, The University of Kansas Medical Center, Kansas City, Kansas, United States
- Rao, Reena, The University of Kansas Medical Center, Kansas City, Kansas, United States
Background
Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations in the PKD1 or PKD2 genes. Progressive renal fibrosis, which often accompanies cyst growth in ADPKD, contributes to the progression to end-stage kidney disease. Myofibroblasts, the primary producers of extracellular matrix (ECM) proteins play a critical role in fibrosis development. While the effect of Pkd1 mutations in tubular epithelium have been extensively charecterized, their impact on interstitial myofibroblasts remains unclear. This study examined the effect of Pkd1 gene mutation in renal fibroblasts.
Methods
We mutated Pkd1 gene (Pkd1KO) in rat renal fibroblasts (NRK-49F cells) using Crispr/Cas12, and compared their cell proliferation, migration, differentiation, metabolism and ECM production to wild-type (WT) cells. Mitochondrial function including oxygen consumption rate, ATP production, and mitochondrial potential, was measured using a Seahorse XF24 extracellular flux analyzer and TMRE staining. Intracellular calcium dynamics were measured using Fura-10 ratiometric dye in response to ATP and Angiotensin II stimulation.
Results
Pkd1KO cells showed increased expression of α-smooth muscle actin (αSMA) indicative of myofibroblast differentiation, along with enhanced cell proliferation, migration and gene expression of ECM proteins. Pkd1KO cells also displayed significantly higher intracellular calcium responses to both ATP and Angiotensin II stimulation compared to WT cells. Additionally, real-time metabolic analysis revealed increased mitochondrial oxidative phosphorylation in Pkd1KO cells, evidenced by higher oxygen consumption, spare respiratory capacity, and ATP production.
Conclusion
Mutation of Pkd1 gene in rat renal fibroblasts spontaneously induces their differentiation into myofibroblasts, enhancing migration, ECM production, Calcium influx, and oxidative phosphorylation. This study highlights a novel role for Pkd1 in regulating fibroblast activation and metabolism, contributing to the pathogenesis of fibrosis in ADPKD. These findings provide a new in vitro model for investigating the interstitial disease mechanisms in ADPKD.
Funding
- NIDDK Support