PUBLICATION
MFSD12 depletion reduces cystine accumulation without improvement in proximal tubular function in experimental models for cystinosis
- Authors
- Bondue, T., Khodaparast, L., Khodaparast, L., Cairoli, S., Goffredo, B.M., Gijsbers, R., van den Heuvel, L., Levtchenko, E.
- ID
- ZDB-PUB-240328-31
- Date
- 2024
- Source
- American journal of physiology. Renal physiology 326(6): F981-F987 (Journal)
- Registered Authors
- Keywords
- MFSD12, cystinosis, renal Fanconi syndrome, zebrafish
- MeSH Terms
-
- Amino Acid Transport Systems, Neutral/genetics
- Amino Acid Transport Systems, Neutral/metabolism
- Animals
- CRISPR-Cas Systems
- Cystine*/metabolism
- Cystinosis*/genetics
- Cystinosis*/metabolism
- Cystinosis*/pathology
- Disease Models, Animal*
- Epithelial Cells/metabolism
- Humans
- Kidney Tubules, Proximal*/metabolism
- Kidney Tubules, Proximal*/pathology
- Zebrafish*/metabolism
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- PubMed
- 38545650 Full text @ Am. J. Physiol. Renal Physiol.
Citation
Bondue, T., Khodaparast, L., Khodaparast, L., Cairoli, S., Goffredo, B.M., Gijsbers, R., van den Heuvel, L., Levtchenko, E. (2024) MFSD12 depletion reduces cystine accumulation without improvement in proximal tubular function in experimental models for cystinosis. American journal of physiology. Renal physiology. 326(6):F981-F987.
Abstract
Cystinosis is an autosomal recessive lysosomal storage disorder, caused by mutations in the CTNS gene, resulting in an absent or altered cystinosin (CTNS) protein. Cystinosin exports cystine out of the lysosome, with a malfunction resulting in cystine accumulation and a defect in other cystinosin-mediated pathways. Cystinosis is a systemic disease, but the kidneys are the first and most severely affected organs. In the kidney, the disease initially manifests as a generalized dysfunction in the proximal tubules (also called renal Fanconi syndrome). MFSD12 is a lysosomal cysteine importer, that directly affects the cystine levels in melanoma cells, HEK293T cells, and cystinosis patient-derived fibroblasts. In this study, we aimed to evaluate MFSD12 mRNA levels in cystinosis patient-derived proximal tubular epithelial cells (ciPTECs) and to study the effect of MFSD12 knockout on cystine levels. We showed similar MFSD12 mRNA expression in patient-derived ciPTECs in comparison to the control cells. CRISPR MFSD12 knockout in a patient-derived ciPTEC (CTNSΔ57kb) resulted in significantly reduced cystine levels. Furthermore, we evaluated proximal tubular reabsorption after injection of mfsd12a translation-blocking morpholino (TB MO) in a ctns-/- zebrafish model. This resulted in decreased cystine levels, but caused a concentration-dependent increase in embryo dysmorphism. Furthermore, the mfsd12a TB MO injection did not improve proximal tubular reabsorption or megalin expression. In conclusion, MFSD12 mRNA depletion reduced cystine levels in both tested models without improvement of the proximal tubular function in the ctns-/- zebrafish embryo. Additionally, the apparent toxicity of higher mfsd12a TB MO concentrations on the zebrafish development, warrants further evaluation.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping