Title | BESTROPHIN1 mutations cause defective chloride conductance in patient stem cell-derived RPE. |
Publication Type | Journal Article |
Year of Publication | 2016 |
Authors | Moshfegh, Yasmin, Velez Gabriel, Li Yao, Bassuk Alexander G., Mahajan Vinit B., and Tsang Stephen H. |
Journal | Hum Mol Genet |
Volume | 25 |
Issue | 13 |
Pagination | 2672-2680 |
Date Published | 2016 07 01 |
ISSN | 1460-2083 |
Keywords | Bestrophins, Calcium Signaling, Chloride Channels, Chlorides, Eye Proteins, Humans, Induced Pluripotent Stem Cells, Mutation, Retinal Pigment Epithelium, Vitelliform Macular Dystrophy |
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. |
DOI | 10.1093/hmg/ddw126 |
Alternate Journal | Hum. Mol. Genet. |
PubMed ID | 27193166 |
PubMed Central ID | PMC5181636 |
Grant List | R21 AG050437 / AG / NIA NIH HHS / United States R01 EY018213 / EY / NEI NIH HHS / United States R01 EY016822 / EY / NEI NIH HHS / United States R01 EY024665 / EY / NEI NIH HHS / United States K08 EY020530 / EY / NEI NIH HHS / United States R01 EY024698 / EY / NEI NIH HHS / United States P30 EY019007 / EY / NEI NIH HHS / United States P30 CA013696 / CA / NCI NIH HHS / United States T32 GM007337 / GM / NIGMS NIH HHS / United States |