Proteome Analysis of Schizophrenia Brain Tissue
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1 Proteome Analysis of Schizophrenia Brain Tissue Daniel Martins-de-Souza Max Planck Institute for Psychiatry Proteomics and Biomarkers München - Germany Laboratory of Neuroscience (LIM-27) Dept of Psychiatry, Faculty of Medicine University of São Paulo, Brazil Laboratory of Proteomics Dept of Biochemistry, State University of Campinas, Brazil
2 Schizophrenia (SCZ) * Chronic and debilitating psychotic disorder * Affects about 1% of world population * Multifactorial disease caused by differential gene and protein expression + environmental factors
3 Why Proteomics? Genomics Knowledge of the complete set of genes to reveal the diseases mechanisms
4 Pre-Genome: Pre-Genome Era A small set of well known genes
5 Post-Genome: Post-Genome Era A comprehensive set of unknown genes and products
6 Same Genome, Different Proteome Neutrofile Neuron Muscle
7 Proteomics The set of expressed proteins of a cell, tissue or organism. The real players
8 Two-Dimensional Gel Electrophoresis (2DE) and Mass Spectrometry (MS) Mass Spectrometry MALDI-TOF
9 Two-Dimensional Gel Electrophoresis (2DE)
10 Shotgun Mass Spectrometry Martins-de-Souza et al. J Neural Transm. 2008
11 Schizophrenia and Proteomics Our goals * Molecular markers for diagnosis * Pathobiology of the disease
12 Investigated Brain Regions Provided by BrainNet Europe Dorsolateral Prefrontal Cortex (BA46) Shotgun-MS and 2DE/MS Confirm differentially expressed genes and proteins Anterior Temporal Lobe (BA38) Novel differentially expressed proteins Shotgun-MS Wernicke s Area (BA22p) 2DE/MS november2007/ brain.htm
13 Dorsolateral Prefrontal Cortex 2DE/MS: 14 upregulated and 10 downregulated proteins Shotgun/MS: 77 upregulated and 7 downregulated proteins
14 Dorsolateral Prefrontal Cortex 2DE/MS and Shotgun-MS a) Energy Metabolism b) Oxidative stress c) Oligodendrocytes metabolism d) Calcium homeostasis e) Immune System-related f) Cytoskeleton Martins-de-Souza et al. J Psychiatr Res in press Martins-de-Souza et al. Eur Arch Psychiatry Clin Neurosci in press
15 Anterior Temporal Lobe Shotgun-MS 6 upregulated and 31 downregulated proteins a) Oligodendrocytes metabolism b) Calcium homeostasis c) Cytoskeleton
16 Wernicke s Area 2DE/MS 14 upregulated and 11 downregulated proteins a) Energy Metabolism b) Cytoskeleton Martins-de-Souza et al. BMC Psychiatry 2008 submitted
17 SCZ Differential Proteome Our results: 19 proteins commonly regulated in the 3 regions of the same patients 10 same direction 9 different direction 15 different biological processes
18 SCZ Differential Proteome Oligodendrocyte Metabolism 3% Immune Response 4% Transport 5% Unknown Biological Process 6% Regulation of nucleobase, nucleoside, nucleotide and nucleic acid metabolism. 7% Biological Processes Revealed in SCZ Osmoregulation/Hormone metabolism Nucleic acid metabolism 1% 1% Synaptic Transmission 2% Neurogenesis 2% DNA repair 1% Regulation of gene expression 1% Metabolism; Energy pathways 23% Cell Growth and/or maintenance 18% Protein Metabolism 9% Signal transduction and Cell communication 17% 15 different biological processes
19 SCZ Differential Proteome Calcium Differential Influx, Immune System and Neurotransmission Dopamine receptors are Ca +2 dependent Glutamine vesicles proteins Immune Systemrelated HSPs and antibodies Jones et al. Immunol Cell Biol Feb;83(1):9-17.
20 SCZ Differential Proteome MBP & MOG & Ermin downregulated Oligodendrocyte Metabolism 2',3'-cyclic-nucleotide 3'- phosphodiesterase (CNP)
21 Biomarker Candidates Martins-de-Souza et al. BMC Psychiatry 2008 submitted
22 SCZ Pathobiology Genetic differences (small effects) DIFFERENTIAL GENE AND PROTEIN EXPRESSION ENVIROMENTAL FACTOR (exogen alteration) NEURODEVELOPMENT (Morphologic alterations) DOPAMINERGIC dysfunction GLUTAMATERGIC dysfunction Altered state of NMDA receptors SEROTONERGIC dysfunction Differential Ca +2 flow MITOCHONDRIAL dysfunction IMMUNE response Neuronal Transmission NO ROS Neuronal and Synaptic Plasticity Myelination systematically altered Apoptosis Cellular Death?
23 Next Steps: * Proteome of different brain regions (Proteome of complete SCZ brain clinical aspects) * Validate the common biomarker candidates in brain tissue, CSF and plasma/serum * Investigation of protein biomarkers in vivo and in vitro * Proteome analysis of naive patient specimens (Prabakaran et al., 2007)
24 Max Planck Institute for Psychiatry München Germany Prof. Dr. Chris W. Turck Dr. Giuseppina Maccarrone Christiane Rewerts Maria Lebar Dr. Jeeva Varadarajulu Larysa Teplytska Stefan Reckow Philipp Gormanns Christian Webhofer Yaoyang Zhang Michaela Filiou Dr. Claudia Ditzen Prof. Dr. Florian Holsboer Laboratory of Neuroscience LIM-27 University of São Paulo, Brazil Dr. Emmanuel Dias-Neto Prof. Dr. Wagner F. Gattaz Laboratory of Proteomics State University of Campinas, Brazil Prof. Dr. José Camillo Novello Prof. Dr. Sérgio Marangoni Georg-August-Universität Göttingen Dept of Neuropathology Dr. Andrea Schmitt Prof. Dr. Peter Falkai
25 The Scream Edvard Munch ( )
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