Abstract: FR-PO586
Modeling the Impact of Polycystic Kidney Disease Genetic Modifiers Using Kidney Organoids Generated from Autologous Induced Pluripotent Stem Cells
Session Information
- Cystic Kidney Diseases: Mechanisms and Models
October 25, 2024 | 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: Cystic
Authors
- Knezevic, Sara, Royal College of Surgeons in Ireland Faculty of Medicine and Health Sciences, Dublin, Ireland
- Davis, Jessie, University College Dublin, Dublin, Ireland
- Elhassan, Elhussein Aamir Elzein, Beaumont Hospital, Dublin, Ireland
- Clince, Michelle, Beaumont Hospital, Dublin, Dublin, Ireland
- Benson, Katherine A., Royal College of Surgeons in Ireland Faculty of Medicine and Health Sciences, Dublin, Ireland
- Conlon, Peter J., Beaumont Hospital, Dublin, Dublin, Ireland
- Crean, John, University College Dublin, Dublin, Ireland
Background
Autosomal dominant polycystic kidney disease (ADPKD) is the most common inherited renal disease caused by pathogenic variants in PKD1 or PKD2. Preliminary evidence for an effect of genetic modifiers (non-pathogenic variants which modify patient phenotype) on disease severity has been reported. Kidney organoids (KO) grown from patient-derived induced pluripotent stem cells (iPSCs) can be used to inform on the variability in disease severity between family members and offer a deeper understanding of ADPKD pathogenesis. Here, iPSCs were generated from an affected individual with ADPKD.
Methods
Patient was recruited from the inherited kidney disease clinic at Beaumont Hospital (Ireland) based on their genetic risk profile. A urine sample was collected and Urinary Epithelial Cells (URECs) were isolated and cultured. URECs were reprogrammed to iPSCs (UiPSC) and validated. KOs were generated from both patient and healthy control UiPSCs and characterised by RT-qPCR and immunocytochemistry. KOs from both patient and healthy control were treated with forskolin to induce cystogenesis and assessed for tubular swelling by brightfield microscopy.
Results
The index patient, a 77-year-old female, was diagnosed with hypertension, a family history of ADPKD and reduced eGFR (35 mL/min) at the last follow-up. Imaging revealed enlarged cystic kidneys and liver cysts indicative of ADPKD. Exome sequencing revealed PKD1 complex variants, a diagnostic variant [c.7300C>T:p.R2434W] and two genetic modifiers [c.C2755T:p.R919W and c.C8293T:p.R2765C]. URECs were successfully isolated from the patient and reprogrammed into UiPSCs. KOs generated from healthy control expressed markers of mature proximal tubule, podocyte, endothelia and stroma. Patient KOs developed cysts alone or with forskolin injury. Tubular abnormalities were evident by Day 9 and more pronounced by Day 13. Healthy control KOs formed cysts only upon forskolin treatment.
Conclusion
Autologous UiPSCs can be used to generate KOs which mimic pathogenic cystogenesis, both spontaneously and post-acute injury. Further comparative studies will examine the mechanisms through which genetic modifiers confer risk and contribute to the severity of the disease, informing future therapeutic approaches to the management of ADPKD.