Disease modeling using induced pluripotent stem cells (iPSC)

Neurodevelopmental and neurodegenerative disorders have an incidence of at least 10% in Western populations. Little is known about the mechanisms and pathophysiology leading to the majority of these central nervous system disorders. To this end, we established induced pluripotent stem cell (iPSC) technology in order to model neurogenesis and neuronal function associated with disease. The objective is to understand disease mechanisms of the central nervous system associated with specific gene variants and, in the long-term, to interfere with these mechanisms in search for novel therapeutic strategies. Patient derived fibroblasts are reprogrammed into iPSC using non-integrating vectors expressing the four Yamanaka factors. Reprogramming is followed by differentiation into neuronal progenitor cells and mature neuronal subpopulations using established protocols. Fibroblasts are obtained from patients with well-defined neurodevelopmental disorders and known causative gene mutations as well as healthy controls. Functional analyses and validation of neuronal cells comprise e.g. proliferation, apoptosis, migration, dendrite formation, electrophysiological properties, imaging techniques and high through-put analysis of biomarkers  (transcriptome, proteome and metabolome analysis) on bulk as well as single cells. Further validations are performed by CRISPR/Cas9 editing of selected genes in iPSC to generate isogenic control/disease cell lines. Disorders that are currently modeled and under investigation include Down syndrome, Alzheimer’s disease, Dravet disease, Mowat-Wilson disease and Von Hippel-Lindau disease. Disease-associated pathways/factors/biomarkers are currently being validated for future screening of small compound libraries.