Abstract
Purpose :
Recently, two research groups discovered that lanosterol and its derivatives had the ability to depolymerize the aggregated proteins and restored partial vision in animal models, which offers great hope in cataracts treatment using eye drops rather than surgery. However, whether lanosterol and its derivatives have similar effects on human cataracts as in animal models are still unclear. Our previous study established an efficient system-“fried egg” method in vitro for lentoid body (LB) generation from human pluripotent stem cells (PSCs), and found that LBs differentiated from congenital cataract (CC) patient specific iPSCs were cloudy which were consistent with the clinic symptoms of the patients (ARVO 2016, 2018). So, this study is to examine whether lanosterol could reverse the opacity of the patient-specific LBs.
Methods :
Patient LBs were derived from CC patients’ iPSCs with β-crystallin,γ-crystallin and unknown mutations. Lanosterol were dissolved in DMSO in a concentration of 4 mM and diluted into 4 mM of working solution. The LB’s morphology was examined at certain time points. Representative images were taken using light microscopy and were used to evaluate the LBs’ degrees of transparency. Immunofluorescence staining was used to examine the expression patterns of crystallin. Representative pictures were used to quantify the number of protein aggregations.
Results :
As the crystallin protein aggregations are the pathological foundation of cataracts, patients’ LBs were treated with lanosterol at D11 of differentiation process when the lens epithelial cells started to differentiate into lens fiber cells with massively increasing of crystalline expression. Our results showed that the percentage of transparent LBs on D21 were significantly higher in lanosterol treated conditions when compared to those without lanosterol treatment in three different types of LBs. Immunofluorescence examination showed that the number of crystallin aggregations was significantly decreased in patients’ LBs treated with lanosterol comparing to those without treatment, which was consistent with soluble/insoluble protein analysis.
Conclusions :
Our results provide evidence that lanosterol is able to reverses the opacity of CC patient specific LBs, indicating the potential use of patient-specific LB platform in cataract drug screening.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.