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Hum Mol Genet. 2016 Jul 1;25(13):2672-2680. Epub 2016 May 18.

BESTROPHIN1 mutations cause defective chloride conductance in patient stem cell-derived RPE.

Author information

1
Barbara & Donald Jonas Laboratory of Regenerative Medicine, and Bernard & Shirlee Brown Glaucoma Laboratory, Department of Pathology & Cell Biology, Institute of Human Nutrition, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
2
Edward S. Harkness Eye Institute, New York-Presbyterian Hospital, New York, NY 10032, USA.
3
Omics Lab.
4
Department of Ophthalmology and Visual Sciences.
5
Medical Scientist Training Program.
6
Department of Pediatrics, University of Iowa, Iowa City, IA, USA.
7
Barbara & Donald Jonas Laboratory of Regenerative Medicine, and Bernard & Shirlee Brown Glaucoma Laboratory, Department of Pathology & Cell Biology, Institute of Human Nutrition, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA sht2@columbia.edu.

Abstract

Bestrophin1 (BEST1) is expressed in human retinal pigment epithelium (RPE) and mutations in the BEST1 gene commonly cause retinal dysfunction and macular degeneration. BEST1 is presumed to assemble into a calcium-activated chloride channel and be involved in chloride transport but there is no direct evidence in live human RPE cells to support this idea. To test whether BEST1 functions as a chloride channel in living tissue, BEST1-mutant RPE (R218H, L234P, A243T) were generated from patient-derived induced pluripotent stem cells and compared with wild-type RPE in a retinal environment, using a biosensor that visualizes calcium-induced chloride ion flux in the cell. Calcium stimulation elicited chloride ion export in normal RPE but not in RPE derived from three patients with BEST1 mutations. These data, along with three-dimensional modeling, provide evidence that BEST1 assembles into a key calcium-sensing chloride channel in human RPE.

PMID:
27193166
PMCID:
PMC5181636
DOI:
10.1093/hmg/ddw126
[Indexed for MEDLINE]
Free PMC Article

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