PUBLICATION
Development of a riboflavin-responsive model of riboflavin transporter deficiency in zebrafish
- Authors
- Choueiri, C.M., Lau, J., O'Connor, E., DiBattista, A., Wong, B.Y., Spendiff, S., Horvath, R., Pena, I., MacKenzie, A., Lochmüller, H.
- ID
- ZDB-PUB-241216-3
- Date
- 2024
- Source
- Human molecular genetics : (Journal)
- Registered Authors
- Horvath, Rita, Pena, Izabella
- Keywords
- CRISPR/Cas9, neurogenetics, riboflavin transporter deficiency, zebrafish
- MeSH Terms
-
- Humans
- Animals
- Larva/genetics
- Larva/metabolism
- Gene Knockdown Techniques
- Bulbar Palsy, Progressive*/drug therapy
- Bulbar Palsy, Progressive*/genetics
- Receptors, G-Protein-Coupled
- Riboflavin*/metabolism
- Riboflavin*/pharmacology
- Probenecid*/pharmacology
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- Mutation
- Zebrafish*/genetics
- Hearing Loss, Sensorineural*/drug therapy
- Hearing Loss, Sensorineural*/genetics
- Hearing Loss, Sensorineural*/metabolism
- Disease Models, Animal*
- Membrane Transport Proteins*/genetics
- Membrane Transport Proteins*/metabolism
- PubMed
- 39656631 Full text @ Hum. Mol. Genet.
Citation
Choueiri, C.M., Lau, J., O'Connor, E., DiBattista, A., Wong, B.Y., Spendiff, S., Horvath, R., Pena, I., MacKenzie, A., Lochmüller, H. (2024) Development of a riboflavin-responsive model of riboflavin transporter deficiency in zebrafish. Human molecular genetics. :.
Abstract
Riboflavin transporter deficiency (RTD) is a rare and progressive neurodegenerative disease resulting from the disruption of RFVT2- and RFVT3- mediated riboflavin transport caused by biallelic mutations in SLC52A2 and SLC52A3, respectively. The resulting impaired mitochondrial metabolism leads to sensorimotor neurodegeneration and symptoms including muscle weakness, respiratory difficulty, and sensorineural deafness. Although over 70% of patients with RTD improve following high-dose riboflavin supplementation, remaining patients either stabilise or continue to deteriorate. This may be due to the rapid excretion of central nervous system (CNS) riboflavin by organic anion transporter 3 (OAT-3), highlighting the need for alternative or supplemental RTD treatments. Probenecid is a promising therapeutic candidate for RTD due to its known inhibitory effect on OAT-3. Therefore, this study aimed to generate morpholino-mediated knockdowns of human SLC52A3 ortholog slc52a3 in zebrafish larvae for use in therapeutic screening of riboflavin and probenecid. Knockdown of slc52a3 resulted in an RTD-like phenotype indicative of altered neurodevelopment, hearing loss, and reduced mobility. This RTD-like phenotype overlaps with the phenotype of CRISPR/Cas9-mediated knockout of slc52a3 in zebrafish, is maintained following slc52a3 morpholino + p53 morpholino co-injection, and is rescued following slc52a3 morpholino + human SLC52A3 mRNA co-injection, indicating specificity of the knockdown. Riboflavin treatment alone ameliorates locomotor activity and hearing ability in slc52a3 morphants. Riboflavin and probenecid co-treatment provides an additional small benefit to hearing but not to locomotion. Our findings demonstrate that this model recapitulates both the RTD phenotype and the riboflavin-responsiveness of RTD patients, and possible therapeutic benefit conferred by probenecid warrants further investigation.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping