|set/DZF Project ProteoSys
|Establishment of in vitro models for neuroprotection with molecular connotations to human neurodegenerative diseases |
Prof. Dr. André Schrattenholz, CSO
General background: Neural derivatives of human and murine embryonic stem cells (hESC and mESC) as potential replacement of animal models for stroke, Alzheimer, multiple sclerosis and ALS.
The aim of this project is the characterization of endpoints from murine and human stem cell models for the quantification of neuroprotective effects of therapeutic compounds. Previous work with these models has so far aimed at a precise molecular and functional description of neurotoxic conditions (protein biomarker signatures for neuronal stress). Based on established protocols for neural differentiation in both models, it is planned to further examine various stages of neuronal maturation with regard to neurotransmitters, known marker proteins and pharmacologic properties, and subsequently to define appropriately correlated endpoints for both species.
Functional endpoints of this type would then be differently screened for surrogate markers by advanced Proteomics technologies. Previous work of the team conducting this study has shown, that the approach adequately represents molecular events and processes underlying certain human diseases of the central nervous system and can be successfully used for the quantification of compound effects (Sommer et al., 2004; Schrattenholz et al., 2005; Schrattenholz & Šoškić, 2006). Briefly, neuronal stress is induced by three different conditions, representing human pathomechanisms, namely ischemia, excitotoxicity and toxicity induced by amyloidogenic peptides. Functional and molecular events observed so far in the murine ESC and other models (Falsig et al., 2006; Lund et al., 2006, Schillo et al., 2005) encourage us to extend these studies to human ESC models, with the ultimate aim to develop a human in vitro system representing crucial aspects of human neurodegenerative diseases, which thus could at least partially replace corresponding animal models like e.g. MCAO-models for stroke, transgenic mouse models for Alzheimer’s disease and ALS, or the extremely irksome models for multiple sclerosis (MOG-EAE).
Falsig J, Pörzgen P, Lund S, Schrattenholz A, Leist M (2006). The inflammatory transcriptome of reactive murine astrocytes and implications for their immune functions. J. Neurochem. 96(3), 893-907.
Lund S, Christensen KV, Hedtjärn M, Mortensen AL, Hagberg H, Falsig J, Hasseldam H, Schrattenholz A, Pörzgen P and Leist M (2006). The dynamics of the LPS triggered inflammatory response of murine microglia under different culture and in vivo conditions. J Neuroimmunol. 2006 Sep 19; [Epub ahead of print].
Schillo S, Pejović V, Hunzinger C, Hansen T, Poznanović S, Kriegsmann J, Schmidt WJ and Schrattenholz A (2005). Integrative Proteomics: functional and molecular characterization of a particular glutamate-related neuregulin isoform. Journal of Proteome Research 4 (3), 900-908.
Schrattenholz A and Šoškić V (2006). NMDA receptors are not alone: Dynamic regulation of NMDA receptor structure and function by neuregulins and transient cholesterol-rich membrane domains leads to disease-specific nuances of glutamate-signalling Current Topics in Medicinal Chemistry, 6(7), 663-686.
Schrattenholz A, Wozny W, Klemm M, Schroer K, Stegmann W, Cahill MA (2005). Differential and Quantitative Molecular Analysis of Ischemia: Complexity reduction by isotopic labeling of proteins using a neural embryonic stem cell model; J. Neurological Sciences, 229-230 (1), 261-267.
Sommer S, Hunzinger C, Schillo S, Klemm M, Biefang-Arndt K, Schwall G, Pütter S, Hoelzer K, Schroer K, Stegmann W Schrattenholz A (2004). Molecular analysis of homocysteic acid-induced neuronal stress. Journal of Proteome Research 3(3), 572-581.