Establishment and analysis of transgenic human embryonic stem cells and from hESCs derived neural stem cells

Coordinator:    Prof. Dr. Oliver Brüstle
Institution: Institut für Rekonstruktive Neurobiologie, Universität Bonn
Mouse embryonic stem cells represent a powerful system for studying molecular mechanisms underlying pathophysiological causes of human disease. However, the potential of modeling neurological and psychiatric disorders is limited due to genetic, developmental, biochemical, and morphological differences between humans and mice. Thus, animal models do not always fully recapitulate the phenotype of the human disorder. The availability of human embryonic stem cells (hESC) provides new perspectives for generating a large variety of human cell types in vitro. In this subproject we will take advantage of hESCs to provide a cellular basis for the systematic mapping of physiological protein-protein interactions in human cells. The project builds on recently established know-how to differentiate hESCs into neural stem cells and further into mature neurons and glia. Specifically, we will employ undifferentiated hESCs well as stably proliferating neural stem cells derived from hESCs (hES-NSCs) recently established in our laboratory. Using these hESC-based model we intend to establish an experimental platform for the analysis of protein-protein interaction networks in human cells. For that we will apply the protein tagging strategy developed by the existing gene trap resource for a set of candidate genes. Here we will initially concentrate on genes involved in Alzheimer's disease, Parkinson disease, schizophrenia and major depression with a particular focus on genes expected to exhibit species-specific differences. Modified transgenic hESCs or hES-NSCs will be differentiated into neurons and glia, which will be subjected to fluorescence imaging to study expression and correct compartmentalization of the tagged proteins. Subsequently, cells will be used for proteomic mapping of tagged genes by high mass accuracy mass spectroscopy. Data will be compared with mouse proteomic data. We expect the species-specific expression of individual disease-related proteins to reveal pathogenic disequilibria not visible in non-human systems.

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