Functional characterization of NGLY1 induced pluripotent stem cell-derived glutamatergic neurons in vitro

Authors

• ZA Shyr
• WF Borschel
• A Farkondeh Kalat
• W Zheng

Abstract

There are an estimated 30 million people affected by a rare disease in the United States alone, but lack of data from basic as well as clinical research continues to be a major hurdle for drug discovery for rare diseases. By applying recent technological advancements, patient-derived induced pluripotent stem cells (iPSCs) from somatic cells can be differentiated into specific cell types and organoids, which enables in vitro examination of disease pathophysiology to identify potential therapeutics. Here we performed screening assays for an ultra-rare genetic disorder caused by deficiency of NGLY1 (N-glycanase), an enzyme that de-glycosylates N-glycans from glycoproteins. NGLY1 deficiency leads to an array of clinical symptoms with predominant neurological impairments. Our goal was to identify suitable phenotypic screens for potential therapeutics for this disease. We examined functional differences between iPSC-derived cortical glutamatergic neurons from that of a healthy donor and two pathogenic NGLY1 mutation variants. Functional assessments made by multi-electrode array analysis, high-throughput measurements of intracellular calcium oscillations, as well as fluorescence imaging suggested that both NGY1 disease lines matured slower in culture and had impairments in neuronal network formation and synchronous activity. This work provides the first steps in developing high-throughput screening assays in a physiologically relevant, patient-derived model of NGYL1 deficiency.

Scientific Focus Area: Stem Cell Biology

This page was last updated on Tuesday, August 6, 2024