HTS with human embryonic stem cells

Induction and characterisation of dopaminergic differentiation by small molecules identified by HTS in human reporter-gene transfected embryonic stem cells and neural progenitor cells.

Cell-based phenotypic assays and pathway screens with small molecule libraries promise to provide extremely useful chemical probes of complex differentiation processes. Small molecules provide exceptionally important tools for probing signalling targets relevant for stem cell differentiation. The identification of small molecules which induce reproducible differentiation of human stem cells will help to elucidate not only the molecular mechanisms of these phenomena, but will ultimately allow controlled differentiation of stem cells and progenitor cells. With advances in high-throughput screening (HTS) we will analyse about 10,000 compounds by using innovative cell-based assays.
A global three-stage analysis approach will be used for embryonic stem cells: (1) the cells will be transfected using those promoters being relevant for the dopaminergic differentiation, (2) these cells will be screened with a library of small molecules testing which drugs are able to induce dopaminergic differentiation and (3) the underlying mechanisms of action of small molecules during differentiation will be analysed by in vitro studies with a special focus on the Wnt pathway.
The project will help to analyze intracellular pathways being driven by the newly characterized small molecules with molecular biological methods. This will enable to transfer results to pre-clinical practice.