دورية أكاديمية
MFSD12 depletion reduces cystine accumulation without improvement in proximal tubular function in experimental models for cystinosis.
| العنوان: | MFSD12 depletion reduces cystine accumulation without improvement in proximal tubular function in experimental models for cystinosis. |
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| المؤلفون: | Bondue T; Laboratory of Pediatric Nephrology, Department of Development and Regeneration, KU Leuven, Leuven, Belgium., Khodaparast L; Switch Laboratory, VIB Center for Brain and Disease Research, Leuven, Belgium.; Switch Laboratory, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium., Khodaparast L; Switch Laboratory, VIB Center for Brain and Disease Research, Leuven, Belgium.; Switch Laboratory, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium., Cairoli S; Laboratory of Metabolic Biochemistry, Department of Pediatric Medicine, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy., Goffredo BM; Laboratory of Metabolic Biochemistry, Department of Pediatric Medicine, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy., Gijsbers R; Laboratory for Molecular Virology and Gene Therapy, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Leuven, Belgium.; Leuven Viral Vector Core, KU Leuven, Leuven, Belgium., van den Heuvel L; Laboratory of Pediatric Nephrology, Department of Development and Regeneration, KU Leuven, Leuven, Belgium.; Department of Pediatric Nephrology, Radboud University Medical Center, Nijmegen, The Netherlands., Levtchenko E; Laboratory of Pediatric Nephrology, Department of Development and Regeneration, KU Leuven, Leuven, Belgium.; Department of Pediatric Nephrology, Emma Children's Hospital, Amsterdam UMC, Amsterdam, The Netherlands. |
| المصدر: | American journal of physiology. Renal physiology [Am J Physiol Renal Physiol] 2024 Jun 01; Vol. 326 (6), pp. F981-F987. Date of Electronic Publication: 2024 Mar 28. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Physiological Society Country of Publication: United States NLM ID: 100901990 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1522-1466 (Electronic) Linking ISSN: 15221466 NLM ISO Abbreviation: Am J Physiol Renal Physiol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Bethesda, Md. : American Physiological Society, c1997- |
| مواضيع طبية MeSH: | Zebrafish*/metabolism , Kidney Tubules, Proximal*/metabolism , Kidney Tubules, Proximal*/pathology , Cystinosis*/metabolism , Cystinosis*/genetics , Cystinosis*/pathology , Cystine*/metabolism , Disease Models, Animal*, Animals ; Humans ; Zebrafish Proteins/metabolism ; Zebrafish Proteins/genetics ; Epithelial Cells/metabolism ; Amino Acid Transport Systems, Neutral/genetics ; Amino Acid Transport Systems, Neutral/metabolism ; CRISPR-Cas Systems |
| مستخلص: | 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 with the control cells. CRISPR MFSD12 knockout in a patient-derived ciPTEC ( CTNS Δ57kb ) 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. In addition, the apparent toxicity of higher mfsd12a TB MO concentrations on the zebrafish development warrants further evaluation. NEW & NOTEWORTHY In this study, we show that MFSD12 depletion with either CRISPR/Cas9-mediated gene editing or a translation-blocking morpholino significantly reduced cystine levels in cystinosis ciPTECs and ctns -/- zebrafish embryos, respectively. However, we observed no improvement in the proximal tubular reabsorption of dextran in the ctns -/- zebrafish embryos injected with mfsd12a translation-blocking morpholino. Furthermore, a negative effect of the mfsd12a morpholino on the zebrafish development warrants further investigation. |
| معلومات مُعتمدة: | 11A7821N Fonds Wetenschappelijk Onderzoek (FWO); 11A7823N Fonds Wetenschappelijk Onderzoek (FWO); 1801120N Fonds Wetenschappelijk Onderzoek (FWO); C14/17/11 KU Leuven (Katholieke Universiteit Leuven); CI-CFUK 2021-02 PI Cystinosis Ireland and the Cystinosis Foundation UK co-funded Research Award 2021 |
| فهرسة مساهمة: | Keywords: MFSD12; cystinosis; renal Fanconi syndrome; zebrafish |
| المشرفين على المادة: | 48TCX9A1VT (Cystine) 0 (Zebrafish Proteins) 0 (Amino Acid Transport Systems, Neutral) 0 (CTNS protein, human) |
| تواريخ الأحداث: | Date Created: 20240328 Date Completed: 20240523 Latest Revision: 20240531 |
| رمز التحديث: | 20240531 |
| DOI: | 10.1152/ajprenal.00014.2024 |
| PMID: | 38545650 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1522-1466 |
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| DOI: | 10.1152/ajprenal.00014.2024 |