LOS DERECHOS Y EL DISCURSO JURIDICO

DISEASE

Principle investigator:

Eleanor Coffey, Ph.D., Academy Research Fellow, Turku Centre for Biotechnology, Åbo Akademi and Turku University, BioCity, Tykistokatu 6B, FI-20521 Turku, Finland. Tel. +358-2-3338605, Fax. +358-2-3338000.

Email: [email protected] Homepage: http://www.btk.fi/index. php?id=1240

Biography:

Eleanor Coffey (b. 1967) graduated from Trinity College Dublin in 1990 and received her Ph.D. from the University of Dundee in 1994. She was awarded a Wellcome Trust fellowship to carry out postdoctoral research in Prof. Karl Åkerman’s laboratory from 1994-1997. In 1997 she founded the Neuronal Signalling group at Åbo Akademi and in 2000 joined Turku Centre for Biotechnology as a group leader in molecular and cellular biology. In addition to running a research group, she directs the Cell Imaging Core at Turku Centre for Biotechnology and coordinates the nordic network on Imaging in Biology and Medicine. She currently holds an Academy of Finland Research Fellow position.

Personnel:

Postdoctoral researcher: Minna Tuittila, Ph.D., Graduate students: Artur Padzik, M.Sc., Justyna Zdrojewska, M.Sc., Emilia Komulainen, M.Sc., Raghu Mysore, M.Sc., Yubao Wang, M.Sc., Undergraduate and exchange students: Hanna Heikelä, Lihua Sun, Agnieszka Bialek

Description of the project:

Neurodegenerative disorders such as Alzheimer’s and Parkinson’s disease as well as stroke are characterised by the irreversible loss of nerve cell function. These diseases for which no cure is known are among the most costly to society. The protein kinase JNK is recognised as a critical player in stroke and neurodegeneration. However exactly how this family of kinases mediates cell death in the brain remains largely unknown. Although targeting of JNK for drug-based therapy is already underway, our understanding of the physiological function of JNK in the brain is in its infancy.

A major challenge for signal transduction therapy is to selectively target the pathological function of signalling molecules without interfering with important physiological roles. To achieve this, our lab established a proteomics-based screen to identify protein kinase substrates and thereby broaden our understanding of kinase function. While we have used this methodology to successfully identify both novel and known substrates for JNK, p38 and PIM kinases (collaboration with Päivi Koskinen), among others (collaboration Erwin Wagner), the main focus of our research is to elucidate the molecular mechanism of JNK and JNK targets in the brain. Identification of novel JNK targets such as SCG10 and MAP2, as well as others under study, has highlighted a critical role for JNK in maintaining microtubule homeostasis and subsequently regulating axodendritic architecture. Identification of the JNK phosphorylation site on kinesin-1 helped characterize a role for

JNK in regulation of fast axonal transport in neurons. We combine biochemical, proteomic, cell biology and imaging methods with neuronal and organotypic cultures as well as transgenic mice to validate kinase targets and elucidate their function. In collaboration with Laurent Nyguen, we have established methods to track radial migration of neurons in the developing telencephalon using 4D imaging. In addition, we are examining dendrite and spine morphology in JNK1-/- brains using lucifer yellow iontophoretic loading followed by quantitative 3D image analysis.

An important finding that we stumbled upon regarding JNK function in the nervous system, was the compartmental segregation of physiological verses pathological JNK function to the cytoplasm and nucleus respectively. By using compartment-targeted peptide inhibitors of JNK, we have shown that nuclear JNK activity is critical for neuronal death in response to trophic deprivation (the type of neuronal death that occurs during brain development) and in response to excitotoxic stimuli (the type of neuronal death that occurs in epilepsy, stroke and contributes to neurodegenerative disorders). Interestingly, although JNK is highly localised to the cytoplasm in neurons, we have shown that cytosolic JNK does not to contribute to these modes of neuronal death. Instead, we find that cytosolic JNK regulates physiological processes that maintain neuritic architecture and regulate migration. These functions of JNK are in turn mediated via cytosol-localised targets, independent of JNK-dependent transcriptional regulation. To realise the therapeutic potential of compartmental targeted JNK inhibitors, we are collaborating with Peter Clarke (University of Lausanne). This study investigates the value of nuclear-targeted peptide inhibitors of JNK as protectants from brain damage that occurs following stroke. By gaining information on how apoptotic and physiological functions of signalling molecules are partitioned within the cell allows selective targeting of inhibitors towards loci where pro-apoptotic events take place. This work provides proof of principal that subcellular targeting of inhibitor molecules provides increased specificity with reduced physiological disturbance.

Funding:

EU 6th _{framework STREP “STRESSPROTECT”, EU 6}th _framework

ToK grant, “GAMIDI”, the Academy of Finland, Åbo Akademi University, Turku University Biomedical Sciences Graduate School, Finnish Graduate School in Neurosciences, Drug Discovery Graduate School, Magnus Ehrnrooth’s Stiftelse, CIMO, Sitra and the Torr Joe och Pentti Borgs Foundation.

Collaborators:

Michael Courtney (University of Kuopio), Tuula Kallunki (Danish Cancer Society), Thomas Herdegen (University of Kiel), Peter Clarke (University of Lausanne), Erwin Wagner (Research Institute of Molecular Pathology), Scott Brady (Univeristy of Illinois at Chicago), Laurent Nguyen (University of Liege), Päivi Koskinen (University of Turku), Aideen Long (Trinity College, Dublin).

Selected Publications:

Morfini, G., You, Y., Pollema, S., Kaminska, A., Pigino, G., Liu, K., Yoshioka, K., Björkblom, B., Coffey, E.T., Bagnato, C., Han, D., Huang, C., Banker, G. and Brady, S.T. (2009) Inhibition of fast axonal transport by pathogenic Huntingtin involves activation of JNK3 and phosphorylation of kinesin-1. Nature Neuroscience, in press

Waetzig, V, Wacker, U, Haeusgen, Björkblom,B, Courtney, M.J., Coffey, E.T. Herdegen, T. (2009) Concurrent protective and destructive signalling of JNK2 in neuroblastoma cells. Cellular Signalling, 21: 873-880.

Naumanen, T., Johansen, L.D., Coffey, E.T., Kallunki, T. (2008) Loss of function of IKAP/ELP1: Could neuronal migration defect underlie familial disautonomia? Cell Adhesion and Migration, 2:236-239. Björkblom B, Vainio JC, Hongisto V, Herdegen T, Courtney MJ, Coffey ET. (2008) All JNKs can kill, but nuclear localization is critical for neuronal death. Journal of Biological Chemistry, 283:19704-13. Hongisto, V., Vainio, J.C., Thompson, R., Courtney, M.J., Coffey, E.T. (2008) The Wnt pool of GSK-3-beta is critical for trophic deprivation induced neuronal death. Molecular and Cellular Biology, 285:1515-27.

Dan Johansen, L., Naumanen, T., Knudsen, A., Westerlund, N., Gromova, I., Junttila, M., Nielsen, C., Bottzauw, T., Tolkovsky, A., Westermarck, J., Coffey, E.T., Jäättelä, M., Kallunki, T. (2008) IKAP localizes to membrane ruffles with filamin A and regulates actin cytoskeleton organization and cell migration. Journal of Cell Science, 121:854-64.

Westerlund, N., Zdrojewska, J., Courtney, M., Coffey, E. (2008) SCG10 as a molecular effector of JNK1: Implications for the therapeutic targeting of JNK in nerve regeneration. Expert Opinion on Therapeutic Targets. 12:31-43. Review.

Semanova, M.M., Mäki-Hokkanen, A.M.J., Cao, C., Komarovski, V., Forsberg, K.M., Koistinaho, M., Coffey, E.T., Courtney, M.J. (2007) Rho mediates calcium-dependent activation of p38a and subsequent excitotoxic cell death. Nature Neuroscience, 10(4):436-443.

Tararuk, R., Östman, N., Li, W., Björkblom, B., Padzik, A., Zdrojewska, J., Hongisto, V., Herdegen, T., Konopka, W., Courtney, M.J., Coffey, E.T. (2006) JNK1 phosphorylation of SCG10 determines microtubule dynamics and axodendritic length. Journal of Cell Biology. 173: 265-277.

Björkblom, B., Östman, N., Hongisto, V., Komarovski, V., Filen, J., Nyman, T., Kallunki, T., Courtney, M., Coffey, E. (2005) Constitutively active cytoplasmic JNK1 is a dominant regulator of dendritic architecture; role of MAP2 as an effector. Journal of Neuroscience. 25: 6350-6361.

Yang, J., Lindahl, M., Lindholm, P., Virtanen, H., Coffey, E., Runeberg-Roos, P., Saarma, M. (2004) PSPN/GFRalpha4 has a significantly weaker capacity than GDNF/GFRalpha1 to recruit RET to rafts, but promotes neuronal survival and neurite outgrowth. FEBS Letters.569: 267-271.

Cao, J., Semenova, M.M., Solovyan, V.T., Han, J., Coffey, E.T., Courtney, M.J. (2004) Distinct requirements for p38alpha and c-Jun N-terminal kinase stress-activated protein kinase s in different forms of apoptotic neuronal death. Journal of Biological Chemistry. 279: 35903-35913.

Hongisto, V., Smeds, N., Brecht, S., Herdegen, T., Courtney, M.J., Coffey, E.T. (2003) Lithium blocks the c-Jun stress response and

protects neurons via its action on glycogen synthase kinase 3. Molecular and Cellular Biology. 23: 6027-6036.

Coffey, E.T., Smiciene, G., Hongisto, V., Cao, J., Brecht, S., Herdegen, T., Courtney, M.J. (2002) c-Jun N-terminal protein kinase (JNK) 2/3 is specifically activated by stress, mediating c-Jun activation, in the presence of constitutive JNK1 activity in cerebellar neurons. Journal of Neuroscience. 22: 4335-4345.

Hietakangas, V., Elo, I., Rosenstrom, H., Coffey, E.T., Kyriakis, J.M., Eriksson, J.E., Sistonen, L. (2001) Activation of the MKK4-JNK pathway during erythroid differentiation of K562 cells is inhibited by the heat shock factor 2-beta isoform. FEBS Letters. 505: 168-172. Coffey, E.T., Hongisto, V., Dickens, M., Davis, R.J. and Courtney, M.J. (2000) Dual roles for c-Jun N-terminal kinase in developmental and stress responses in cerebellar granule neurons. Journal of Neuroscience. 20: 7602-7613.

Courtney, M.J. and Coffey, E.T. (1999) The mechanisms of ARA-C induced apoptosis of differentiating cerebellar granule neurons. European Journal of Neuroscience. 11: 1073-1084.

Coffey, E.T. and Courtney, M.J. (1997) Regulation of SAPKs in CNS neurons. Biochem Soc Trans. 25: S568.

Courtney, M.J., Åkerman, K.E.O. and Coffey, E.T. (1997) Neurotrophins protect cultured cerebellar granule cells against the early phase of cell death by a two-component mechanism. Journal of Neuroscience. 17: 4201-4211.

From left to right: Nina Westerlund, Jenni Vainio, Justyna Zdrojevska, Emilia Komulainen, Vesa Hongisto, Carolina Salenius, Benny Björkblom, Artur Padzik and

ORGANISATION OF NEURONAL