Abstract
Congenital cataracts are the leading cause of childhood blindness. To date, surgical removal of cataracts is the only established treatment, but surgery is associated with multiple complications, which often lead to visual impairment. Therefore, mechanistic studies and drug-candidate screening have been intrigued by the aims of developing novel therapeutic strategies. However, these studies have been hampered by a lack of an appropriate human-disease model of congenital cataracts. Herein, we report the establishment of a human congenital cataract in vitro model through differentiation of patient-specific induced pluripotent stem cells (iPSCs) into regenerated lenses. The regenerated lenses derived from patient-specific iPSCs with known causative mutations of congenital cataracts (CRYBB2 [p. P24T] and CRYGD [p. Q155X]) showed obvious opacification that closely resembled that seen in patients’ cataracts in terms of opacification severity and disease course accordingly, as compared with lentoid bodies (LBs) derived from healthy individuals. Increased protein aggregation and decreased protein solubility corresponding to the patients’ cataract severity were observed in the patient-specific LBs and were attenuated by lanosterol treatment. Taken together, the in vitro model described herein, which recapitulates patient-specific clinical manifestations of congenital cataracts and protein aggregation in patient-specific LBs, provides a robust system for research on the pathological mechanisms of cataracts and screening of drug candidates for cataract treatment.
Highlights
Human-induced pluripotent stem cells provide a promising resource to establish human-disease models with individualspecific genetic backgrounds for use in pathological, pharmaceutical, and regeneration studies
The results revealed that the TR% values of the CRYGD-mutated lentoid bodies (LBs) (0.57 ± 1.35%) and the CRYBB2-mutated LBs (34.76 ± 10.85%) were significantly smaller than those of the normal LBs (84.34 ± 7.00%) (p < 0.01) (Fig. 2c)
Considering the difference in the progression between the two patients, these findings indicated that the differentiation processes in these patientspecific LBs were similar to the disease courses of congenital cataracts caused by the corresponding mutations, respectively
Summary
Human-induced pluripotent stem cells (iPSCs) provide a promising resource to establish human-disease models with individualspecific genetic backgrounds for use in pathological, pharmaceutical, and regeneration studies. Models of various diseases, such as cardiac arrhythmias, cardiomyopathy, inherited metabolic liver disorders, diabetes, and Alzheimer’s disease, have been developed based on the differentiation of patient-specific iPSCs into various cells, including cardiac myocytes, hepatocytes, pancreatic betacells, and neurons or tissues[1,2,3,4,5]. These disease models simulate patient-specific phenotypes, having wide applications in the study of pathological mechanisms and the screening of drug candidates under the patient-specific genetic backgrounds. We made several modifications to a previously established protocol of lens regeneration in vitro, the “three-stage” protocol[12], and established a “fried egg” method to generate LBs from human urinary cell-derived iPSCs (UiPSCs)[13]
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