Abstract: PO1353
Adult Zebrafish as a Model to Study Renal and Extrarenal Manifestations of Cystinosis
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
- Berlingerio, Sante Princiero, Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- He, Junling, Universiteit Leiden, Leiden, Zuid-Holland, Netherlands
- Cairoli, Sara, Ospedale Pediatrico Bambino Gesu, Roma, Lazio, Italy
- Goffredo, Bianca Maria, Ospedale Pediatrico Bambino Gesu, Roma, Lazio, Italy
- Van Den Heuvel, L.P.W.J., Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
- Levtchenko, Elena N., Katholieke Universiteit Leuven Groep Biomedische Wetenschappen, Leuven, Flanders, Belgium
Group or Team Name
- Pediatric Nephrology
Background
Cystinosis is a rare autosomal recessive disease caused by mutations in the CTNS gene, which encodes for the cystine transporter cystinosin leading to lysosomal cystine accumulation in all cells of the body, with kidneys being the first affected organs. The current treatment with cysteamine decreases the lysosomal cystine accumulation, but does not reverse the renal Fanconi syndrome, glomerular injury or loss of renal function. We have developed a zebrafish larvae model having truncating mutation in ctns, which recapitulates the kidney phenotype of cystinosis. However, long-term disease consequences in adult zebrafish have not been studied so far. In this study, we characterized the adult zebrafish model to evaluate the late effects of cystinosis on kidney and extra renal organs.
Methods
Cystinosis (ctns- /-) zebrafish of 18 months and wild type (WT) zebrafish were studied. Histologic examinations of kidneys and extra-renal organs were performed. Cleaved caspase-3 staining was used to evaluate apoptosis in the kidney. Cystine accumulation was evaluated via liquid chromatography-mass spectrometry and toluidine blue staining. For the fertility studies, the number of total eggs and fertile eggs produced by breeding female and male ctns- /- zebrafish compared to WT zebrafish was evaluated.
Results
ctns-/- zebrafish show increased cystine level, glomerular hypertrophy and proximal tubular accumulation of hyaline-like eosinophilic droplets and vacuolated cytoplasm. Moreover, the cystinotic zebrafish exhibit increased cleaved caspase-3, indicating enhanced apoptosis in the proximal tubules. In addition, instead of the typical striped pattern, ctns-/- zebrafish present an altered melanin skin pigmentation, resulting in spotted skin. Lastly, male ctns-/- zebrafish show spermatogenic cysts enriched in spermatozoa, while female display increased percentage of unfertilized eggs.
Conclusion
The adult ctns-/- zebrafish model reproduces several phenotypes of cystinosis, such as altered glomerular and proximal tubular morphology, whole body cystine accumulation, impaired skin pigmentation and decreased fertility. Therefore, this model may be useful for studying long-term effects of cystinosis and for the development of new therapeutic strategies for correcting cystinosis, which is - up to now - incurable.