Induced Pluripotent Stem Cell Modeling of Best Disease and Autosomal Recessive Bestrophinopathy

다국어 초록 (Multilingual Abstract)

Purpose: To understand the pathophysiology of Best disease (BD) and autosomal recessive bestrophinopathy (ARB) by establishingan in vitro model using human induced pluripotent stem cell (iPSC).
Materials and Methods: Human iPSC lines were generated from mononuclear cells in peripheral blood of one ARB patient, oneautosomal dominant BD patient, and two normal controls. Immunocytochemistry and reverse transcriptase polymerase chain reactionin iPSC lines were conducted to demonstrate the pluripotent markers. After the differentiation of iPSC into functional retinalpigment epithelium (RPE), morphological characteristics of the RPE were evaluated using confocal microscopy and immunocytochemistry.
The rates of fluid flow across iPSC-RPE monolayer were measured to compare apical to basal fluid transportsby RPE. RNA sequencing was performed on iPSC-RPE to identify the differences in gene expression profiles, and specific genesets were tested using Gene Set Enrichment Analysis.
Results: Morphological characteristics, gene expression, and epithelial integrity of ARB iPSC were comparable to those of BD patientor normal control. Fluid transport from apical to basal was significantly decreased in ARB iPSC-RPE compared with BD iPSCRPEor control iPSC-RPE. Gene Set Enrichment Analysis confirmed that ARB iPSC-RPE exhibited significant enrichments of epithelial-mesenchymal transition gene set and TNF-α signaling via NF-κB gene set compared to control iPSC-RPE or BD iPSC-RPE.
Conclusion: A human iPSC model of ARB showed a functional deficiency rather than anatomical defects. ARB may be caused byRPE dysfunction following BEST1 mutation.

참고문헌 (Reference)

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