Abstract: PO1354
Urine-Derived Kidney Progenitor Cells in Cystinosis: Potential for Disease Modeling and Ex Vivo Gene Therapy
Session Information
- Genetic Diseases of the Kidneys: Non-Cystic - II
November 04, 2021 | Location: On-Demand, Virtual Only
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1002 Genetic Diseases of the Kidneys: Non-Cystic
Authors
- Veys, Koenraad, Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- Berlingerio, Sante Princiero, Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- Bondue, Tjessa, Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- Held, Katharina, Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- Reda, Ahmed, Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- Theunis, Koen, Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- Baatsen, Pieter, Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- Janssen, Mirian Ch, Radboudumc, Nijmegen, Gelderland, Netherlands
- Cornelissen, Elisabeth A.M., Radboudumc, Nijmegen, Gelderland, Netherlands
- Vriens, Joris, Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- Diomedi-Camassei, Francesca, Ospedale Pediatrico Bambino Gesu, Roma, Lazio, Italy
- Gijsbers, Rik, Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- Van Den Heuvel, L.P.W.J., Radboudumc, Nijmegen, Gelderland, Netherlands
- Arcolino, Fanny Oliveira, Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- Levtchenko, Elena N., Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
Background
Nephropathic cystinosis is an inherited multisystem lysosomal storage disorder caused by mutations in the CTNS gene. The kidney phenotype is characterized by excessive shedding of proximal tubular cells and podocytes into urine, development of renal Fanconi syndrome and progression to end-stage kidney disease. We hypothesized that, to compensate for epithelial cell losses, cystinosis kidneys undertake a regenerative effort, albeit maladaptive. We aimed to search for the presence of kidney progenitor cells (KPCs) in urine of cystinosis patients, and to explore the feasibility of ex vivo gene therapy.
Methods
We isolated undifferentiated cells from urine of cystinosis patients, characterized them as KPCs (Cys-uKPCs) and differentiated these to podocytes (Cys-uKPC-Podo) and proximal tubular epithelial cells (Cys-uKPC-PTEC) as shown by qPCR, RNA sequencing, immunostainings and specific functional assays. Complementation of CTNS in Cys-uKPCs was performed via lentiviral vector (LV) transduction.
Results
Cystinosis patients voided high numbers of undifferentiated cells in urine, of which specific clones expressed several kidney progenitor markers, showed a high level of self renewal, and could differentiate into functional podocytes and PTECs. RNA sequencing demonstrated distinctive transcriptomic signatures distinguishing Cys-uKPCs, Cys-uKPC-Podo and Cys-uKPC-PTEC. Ex vivo gene therapy using a gene addition approach with wild-type CTNS showed significant reductions of cystine levels and altered the perinuclear distribution of the LAMP1+ endo-lysosomal compartment.
Conclusion
Kidney progenitor cells are present in the urine of cystinosis patients. These cells can be isolated, differentiated to functional kidney epithelial cells and complemented with wild-type CTNS to improve the cellular phenotype. Cystinosis uKPCs are a novel tool for disease modeling, while we provided proof of principle of ex vivo gene therapy.
Funding
- Private Foundation Support