Abstract
Purpose :
Alström syndrome (AS) is a rare multisystemic monogenic recessive disorder characterized by hearing and vision loss, obesity, type 2 diabetes mellitus, dilated cardiomyopathy, and progressive and renal dysfunction. Visual symptoms develop within a few weeks after birth, progressively leading to blindness. Currently, no treatment is available.
AS is caused by mutations in the ALMS1 gene. The protein coded by this gene localizes at the centrosome and within the basal bodies of ciliated cells and has suggested roles in cell cycle regulation, intraciliary transport, cell migration, extracellular matrix production, and in endosomal trafficking.
Methods :
Our work aims to model cell lines based on hPSCs in order to understand the molecular mechanisms underlying AS and to identify novel therapeutic targets. To this end, the first step is the induction of selected non-sense mutations within ALMS1’s sequence using novel base editing techniques. Simultaneously, KO cell lines are generated using CRISPR Cas9 system.
Results :
The correct induction of the mutations within the targeted sequence of ALMS1 was evaluated in the different cell lines and the effects of these mutations were analyzed with immunofluorescence and proliferation assays. We also assessed the pluripotency of the cell lines with cytometry and immunofluorescence.
Conclusions :
These model hPSC lines were differentiated into retinal cell types like retinal-pigmented epithelium cells and in retinal organoids.
Future work on these differentiated cells will allow to identify the mechanisms involved in the eye-related symptoms of AS.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.