SP4: Modeling dystonia in an endogenous human cellular system: Characterization of iPS C-derived neurons from patients with monogenic isolated dystonia
The research covered in this project is twofold, first to build up a repository for biomaterial from dystonia patients and second to use this material for disease modeling. The biorepository will contain skin fibroblasts and stem cells from patients with different forms of genetic and non-genetic dystonia. We will employ a new technique by using patient-derived skin cells that are reprogrammed into induced pluripotent stem cells (iPSCs).
We intend to differentiate iPSCs from a specific subset of patients with monogen ic isolated dystonia into the diseased cell type, dopaminergic and striatal neurons.
These neurons will be investigated by electrophysiological measurements that will lead to a better understanding of the disease biology and may allow for the development of drug screening assays.
The objectives are:
- To build up a biorepository of fibroblasts and iPSC lines from patients with different forms of genetic and non-genetic dystonia
- To differentiate iPSCs from patients with TOR1A (DYT1), THAP1 (DYT6), and GNAL (DYT25) mutations into dopaminergic and striatal neurons
- To characterize these neurons in a multimodal fashion (calcium imaging, electrophysiology)
Figure: The steps towards human cellular disease models by using induced pluripotent stem cells (iPS cells).
The generation of human cellular disease models that can be used to understand the pathological mechanisms of dystonia and to develop new therapies is divided into four steps: first, isolating and culturing skin cells that are obtained from a patient. Second, the reprogramming of these cells into a pluripotent state (iPS cells). Third, the directed differentiation of those patient-specific pluripotent cells into the cell type relevant to their disease, i.e. specific subtypes of neurons affected in dystonia. And fourth, the examination of these neurons for disease-related phenotypes.