Identification and Correction of Mechanisms Underlying Inherited Blindness in Human iPSC-Derived Optic Cups

Abstract

SummaryLeber congenital amaurosis (LCA) is an inherited retinal dystrophy that causes childhood blindness. Photoreceptors are especially sensitive to an intronic mutation in the cilia-related gene CEP290, which causes missplicing and premature termination, but the basis of this sensitivity is unclear. Here, we generated differentiated photoreceptors in three-dimensional optic cups and retinal pigment epithelium (RPE) from iPSCs with this common CEP290 mutation to investigate disease mechanisms and evaluate candidate therapies. iPSCs differentiated normally into RPE and optic cups, despite abnormal CEP290 splicing and cilia defects. The highest levels of aberrant splicing and cilia defects were observed in optic cups, explaining the retinal-specific manifestation of this CEP290 mutation. Treating optic cups with an antisense morpholino effectively blocked aberrant splicing and restored expression of full-length CEP290, restoring normal cilia-based protein trafficking. These results provide a mechanistic understanding of the retina-specific phenotypes in CEP290 LCA patients and potential strategies for therapeutic intervention.

Eupatilin rescues ciliary transition zone defects to ameliorate ciliopathy-related phenotypes

“…CEP290 gene augmentation therapy using viral vectors has been shown to ameliorate LCA-related phenotypes in human cell and mouse models (7,8). Oligonucleotide-mediated splicing correction has also been shown to rescue LCA phenotypes in the induced pluripotent stem cell-derived optic cups of patients harboring a CEP290 intronic mutation (9). In this study, we aimed to identify lead compounds for the treatment of CEP290-related ciliopathies through cellbased compound library screening.…”

Stem cells with a view: a look inside a retinal ciliopathy

“…They found that the CEP290-MO reduced the levels of the cryptic exon inclusion and resulted in a concomitant increase in the normal transcript levels in LCA fibroblasts. The CEP290-MO treatment also restored ciliation defects and ciliary protein trafficking of RPGR, and of opsins in the 3D-optic cup derived photoreceptors (13).…”

“…In a recent study, Parfitt et al (13) took on this challenging question. They resorted to induced pluripotent stem cells (iPSCs) to not only investigate the disease mechanisms but also to develop rational therapeutic modalities.…”

Personalized models reveal mechanistic and therapeutic insights into CEP290-associated Leber congenital amaurosis

“…In the present report, Parfitt et al generate iPSC-derived models to extend our understanding of the disease process accompanying the most common CEP290 mutation found within the human population (6). The mutation (c.2991 + 1665A>G) creates a cryptic splice donor site within intron 26 of the gene, leading to the inclusion of an additional exon in the mature mRNA sequence.…”