Investigation and Restoration of BEST1 Activity in Patient-derived RPEs with Dominant Mutations
| Autor: | Alec Kittredge, Yu Zhang, Tingting Yang, Nancy Ward, Peng Yao, Changyi Ji, Yao Li, Austin Hopiavuori, Yohta Fukuda, Stephen H. Tsang |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Adult Male Genetic enhancement lcsh:Medicine Retinal Pigment Epithelium Biology Virus Article 03 medical and health sciences chemistry.chemical_compound Young Adult 0302 clinical medicine Gene therapy medicine Humans Bestrophins lcsh:Science Child Gene Genes Dominant Chloride channels Genetics Multidisciplinary Retinal pigment epithelium lcsh:R HEK 293 cells Retinal Hereditary eye disease Middle Aged Subcellular localization Phenotype Retinal diseases Vitelliform Macular Dystrophy 030104 developmental biology medicine.anatomical_structure chemistry Mutation lcsh:Q Female Patch clamp 030217 neurology & neurosurgery |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-13 (2019) |
| ISSN: | 2045-2322 |
| Popis: | BEST1 is a Ca2+-activated Cl− channel predominantly expressed in retinal pigment epithelium (RPE), and over 250 genetic mutations in the BEST1 gene have been identified to cause retinal degenerative disorders generally known as bestrophinopathies. As most BEST1 mutations are autosomal dominant, it is of great biomedical interest to determine their disease-causing mechanisms and the therapeutic potential of gene therapy. Here, we characterized six Best vitelliform macular dystrophy (BVMD)-associated BEST1 dominant mutations by documenting the patients’ phenotypes, examining the subcellular localization of endogenous BEST1 and surface Ca2+-dependent Cl− currents in patient-derived RPEs, and analyzing the functional influences of these mutations on BEST1 in HEK293 cells. We found that all six mutations are loss-of-function with different levels and types of deficiencies, and further demonstrated the restoration of Ca2+-dependent Cl− currents in patient-derived RPE cells by WT BEST1 gene supplementation. Importantly, BEST1 dominant and recessive mutations are both rescuable at a similar efficacy by gene augmentation via adeno-associated virus (AAV), providing a proof-of-concept for curing the vast majority of bestrophinopathies. |
| Databáze: | OpenAIRE |
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