Abstract
Purpose :
Purpose: Leber congenital amaurosis (LCA) is a group of highly heterogeneous, early onset, severe retinal dystrophy. The pathogenicity has not been directly clarified due to lack of diseased cells. This study is to produce and identify RPE65 patient specific induced pluripotent stem cells (iPSCs) and their retinal tissues for disease modeling and drug screening.
Methods :
Urine cells from a patient carrying two novel RPE65 mutations were reprogrammed into iPSCs. The identifiable iPSCs colonies were subjected to stringent characterization, and then guided to differentiate into retinal organoids with our published protocols (Zhong X, Nature Communications 2014; Li G, Stem Cells International 2018).
Results :
LCA patient hiPSCs with two novel RPE65 mutations c.200T>G and c.430T>C were generated. They presented typical morphological features with normal karyotype, expressed pluripotency markers, and developed teratoma in NOD-SCID mice. Moreover, the patient hiPSCs were able to differentiate towards retinal lineage fate and self-form retinal organoids with layered neural retina and retinal pigment epithelium (RPE). All major retinal cell types including photoreceptors were also acquired overtime. Compared to healthy control, RPE cells from patient iPSCs had lower expression of RPE65.
Conclusions :
This study provided the valuable patient specific, disease targeted retinal organoids containing photoreceptor and RPE cells, facilitating the study of personalized pathogenic mechanisms and gene or drug screening.
This is a 2020 ARVO Annual Meeting abstract.