Laboratory of Applied Proteome Analyses

Laboratory of Applied Proteome Analyses

Head of the Laboratory:

Hana Kovarova, Dr.

kovarova@iapg.cas.cz

 

A small group focused on proteomics evolved as a part of the Laboratory of Biochemistry and Molecular Biology of Germ Cells at the Institute of Animal Physiology and Genetics, Czech Academy of Science (IAPG CAS) in Libechov in 2002. The initial focus on maturation and activation of mammalian oocyte was later complemented by general reproductive biology [1–8]. Additionally, cancer research studies including the effects and biochemical mechanisms of anti-cancer drugs and drug resistance facilitated the establishment of the Centre for Cancer Proteomics [9–14] in collaboration with the Laboratory of Experimental Medicine, Palacky Univesity and University Hospital in Olomouc and with the Laboratory of Molecular Structure Characterization, Institute of Microbiology CAS in Prague. Another cancer research aspect of epithelial-mesenchymal cell communication [15, 16] was added thanks to collaboration with the group of Prof. Karel Smetana Jr. at the 1st Faculty of Medicine, Charles University in Prague. In 2006, exciting studies in the field of stem cell biology, namely differentiation of neural stem cells and neural precursor cells, have been initiated in co-operation with the Laboratory of Prof. Martin Marsala at the Department of Anesthesiology, University of California in San Diego [17–21]. Coupled with the therapeutic potential of stem cells and their possible application in neurological or neurodegenerative disorders [22], comprehensive proteome analyses of such cells now represents the key interest of our Laboratory. To monitor the effects of such therapy, we plan to characterize animal models of spinal cord injury [22] as well as a transgenic minipig model of Huntington’s disease recently generated at our Institute. Proteomics provides tool for discovery of biological processes linked both physiological and pathological stages. Proteins participating in these pathways could serve as the biomarkers for observation of disease progression and the efficacy of a potential therapy.

 

Activities

A small group focused on proteomics evolved as a part of the Laboratory of Biochemistry and Molecular Biology of Germ Cells at the Institute of Animal Physiology and Genetics, Czech Academy of Science (IAPG CAS) in Libechov in 2002. The initial focus on maturation and activation of mammalian oocyte was later complemented by general reproductive biology [1–8]. Additionally, cancer research studies including the effects and biochemical mechanisms of anti-cancer drugs and drug resistance facilitated the establishment of the Centre for Cancer Proteomics [9–14] in collaboration with the Laboratory of Experimental Medicine, Palacky Univesity and University Hospital in Olomouc and with the Laboratory of Molecular Structure Characterization, Institute of Microbiology CAS in Prague. Another cancer research aspect of epithelial-mesenchymal cell communication [15, 16] was added to our studies thanks to collaboration with the group of Prof. Karel Smetana Jr. at the 1st Faculty of Medicine, Charles University in Prague. In 2006, exciting studies in the field of stem cell biology, namely differentiation of neural stem cells and neural precursor cells, have been initiated in co-operation with the Laboratory of Prof. Martin Marsala at the Department of Anesthesiology, University of California in San Diego [17–21]. Coupled with the therapeutic potential of stem cells and their possible application in neurological or neurodegenerative disorders [22], comprehensive proteome analyses of such cells now represents the key interest of our Laboratory. To monitor the effects of such therapy, we plan to characterize animal models of spinal cord injury [22] as well as a transgenic minipig model of Huntington’s disease recently generated at our Institute.

 

Proteomics provides tool for discovery of biological processes linked both physiological and pathological stages. Proteins participating in these pathways could serve as the biomarkers for observation of disease progression and the efficacy of a potential therapy.

 

 

Neural stem cell differentiation

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To facilitate transition towards to therapeutic use, the aim of this study is comprehensive characterization of neural differentiation at the level of protein changes with focus on:

a) surface membrane proteins to select cells of desired phenotype

b) intracellular proteins as a readout for developmental stage

c) secreted proteins for modulation of stem cells niche

 

Huntington’s disease and neurodegeneration

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The aim of this project is uncovering of the differences at protein level which are linked to progressing Huntington’s disease and neurodegeneration with focus on:

a) plasma and cerebrospinal fluid proteome alterations

b) immune response including complement system and cytokine analysis

 

Equipment

Luminex 200 Bead based instrument for multiplex immunoassays

Leica SP5 confocal microscope

BD FACSAria cell sorter

AB Sciex TripleTOF 5600+ mass spectrometer coupled to Eksigent nanoLC 425

AB Sciex QTRAP 5500 mass spectrometer coupled to Eksigent nanoLC 425

2D gel electrophoresis and 2D HPLC for protein fractionation

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