Establishing an in vitro human Leber congenital amaurosis model system

摘要: Purpose: Dominant cone-rod homeobox (CRX)-associated Leber congenital amaurosis (LCA7) is a severe retinal degenerative disease for which no treatments are currently available. Disease-causing variants in CRX typically result in the production of a dominant negative form of the protein, which disrupts normal photoreceptor maturation. To gain further insight into LCA7, we aimed to establish an in vitro model system to investigate CRX variant-specific disease mechanisms.Methods: Human iPSC lines were generated from LCA7 patient whole blood samples, which harbor dominant mutations in CRX (CRX T155ins4/+ and CRX K88Q/+). The patient-derived hiPSC lines were differentiated to generate retinal organoids, and immunohistochemistry, TEM, qPCR, and single-cell RNA sequencing were utilized to characterize photoreceptor cell development and maturation up to day 240 (D240) of differentiation …