Abstract: FR-PO573
Generation of a New Mouse Model Harboring the Polycystin-2 Loss-of-Function D511V Patient Variant
Session Information
- Genetic Diseases: Cystic - Basic
November 03, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1201 Genetic Diseases of the Kidneys: Cystic
Authors
- Outeda, Patricia, Deparment of Medicine/Division of Nephrology, University of Maryland, School of Medicine, Baltimore, Maryland, United States
- Haycraft, Courtney J., Department of Cell, Developmental and Integrative Biology, Heersink School of Medicine, Birmingham, Alabama, United States
- Basquin, Denis, Deparment of Medicine/Division of Nephrology, University of Maryland, School of Medicine, Baltimore, Maryland, United States
- Tran, Pamela Vivian, The Jared Grantham Kidney Institute, University of Kansas Medical Center, Kansas City, Kansas, United States
- Parnell, Stephen C., The Jared Grantham Kidney Institute, University of Kansas Medical Center, Kansas City, Kansas, United States
- DeCaen, Paul G., Department of Pharmacology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, United States
- Wallace, Darren P., The Jared Grantham Kidney Institute, University of Kansas Medical Center, Kansas City, Kansas, United States
- Yoder, Bradley K., Department of Cell, Developmental and Integrative Biology, Heersink School of Medicine, Birmingham, Alabama, United States
- Watnick, Terry J., Deparment of Medicine/Division of Nephrology, University of Maryland, School of Medicine, Baltimore, Maryland, United States
Group or Team Name
- On Behalf of the Polycystic Kidney Disease Research Resource Consortium (PKD-RRC).
Background
Autosomal Dominant Polycystic Kidney Disease (ADPKD) is one of the most frequent renal genetic disorders caused by genetic mutations in either PKD1 or PKD2 genes, which encodes for protein products polycystin-1 (PC1) and polycystin-2 (PC2), respectively. The molecular pathogenesis of ADPKD and the function of polycystins is an area of active research. PC2 subunits can form ion channels within organelle membranes such as the ER and primary cilia, or it can form channel complexes with PC1. There remains a need to develop novel animal models to mimic mutations observed in humans to study the allelic contributions of PKD genes, and to understand the impact of ADPKD-causing variants. To date there are no mouse models with Pkd2 missense variants.
Methods
We used CRISPR/Cas9 gene editing to engineer a new Pkd2 mouse model with the human pathogenic variant D511V (D509V in the mouse, Pkd2D509V). This Pkd2 variant affects a highly conserved region in the voltage-sensor domain (VSD). Prior studies in cell models showed that PKD2D511V encodes a temperature sensitive mutant protein that is more stable at 27°C versus 37°C, interacts with PC1 and can traffic to cilia. Nonetheless this protein is “channel-dead” in ER vesicles.
Results
We found that heterozygous Pkd2D509V mice are viable and fertile but initial studies suggest that homozygous carriers are embryonically lethal, as no homozygotes were alive at P0 (postnatal day 0). We are in the process of determining the cause of embryonic lethality and of measuring PC2D509V protein abundance and ciliary localization “in vivo” and “in vitro”. We will examine the functional role of the PC2D509V variant in maintaining kidney homeostasis in adult and postnatal kidneys by crossing it to a renal tubule specific inducible Pkd2 mouse model.
Conclusion
The generation of this new Pkd2D509V model provides a relatively rapid system to: 1) study the roles of PC2 “in vivo”; 2) decipher the pathogenic mechanisms leading to cyst formation after inactivating the channel activity of PC2 and 3) may be useful for precision medicine applications using patient-derived variants.
Funding
- NIDDK Support