In Parkinson Disease (PD) the content of fibroblast growth factor-2 (FGF-2) in dopamine neurons in substantia nigra pars compacta (Snc) becomes depleted prior to cell degeneration. In the Snc the most abundant are the HMW non-released isoforms of FGF-2 that localize almost exclusively in the cell nucleus. Functions of nuclear FGF-2 and the significance of its depletion in Snc neurons in PD patients remain unknown. We have described a new Integrative Nuclear FGF41 Signaling (INFS) pathway through which diverse extracellular factors and intracellular cAMP and Protein Kinase C regulate growth and survival of neuronal cells and proliferation of astrocytes. These broad functions are likely to involve regulation of several gene programs. This INFS is initiated by the activation of the FGF-2 and FGF receptor-1 (FGFR1) genes followed by transfer of FGFR1 and FGF-2 proteins directly from the cytoplasm to the cell nucleus: Nuclear FGFR1, interacts with Ribosomal S6 Kinase-1 (RSK-1) and stimulate transcription of different genes. The INFS regulates axonal and dendritic growth in cultured neurons and resistance to 6-hydroxydopamine in PC12 cells. Polyetheleneimine-mediated in vivo transfection of dominant negative of the FGFR1 mutant [FGFR1(TK-) lacking tyrosine kinase domain] into the rat Snc caused a delayed loss of tyrosine hydrosxylase expressing neurons and depletion of striatal dopamine. This indicated that TGFR1 signaling is essential also for the long term survival of DA neurons and impaired INFS (i.e. due to a depletion of nuclear FGF-2) may be a common underlying cause for the neuronal degeneration in PD. To elucidate the role of INFS in survival regeneration and functional plasticity of the rat Snc DA neurons, we will use PEI-mediated transfection of FGFR1 and RSK-1 mutants, in vivo, into the adult rat brain. These experiments will establish a new, gene- transfer-based model for common PD and may identify new molecular targets for gene-transfer and pharmacological therapies for the PD and related neurodegenerative disorders.
The specific Aims are: (1) To determine time courser and characterize neurodegenerative changes in the nigrostriatal pathway caused by transfection of FGFR (TK-) and establish whether impaired FGFR signaling directly in the cell nucleus is responsible for the observed effects of FGFR1 (TK-), (2) To identify types of genes regulated by nuclear FGFR1 in rat Snc and whether over- expression of active RSK-1 counteracts the FGFR1(TK)-induced changes in gene expression and degeneration of the nigrostriatal pathway, and (3) To determine whether direct stimulation of nuclear FGFR-1 signaling may attenuate degenerative changes in the nigrostriatal pathway caused by neurotoxin 6-hydroxydopamine and/or promote regeneration.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21NS043621-02
Application #
6625880
Study Section
Special Emphasis Panel (ZNS1-SRB-K (03))
Program Officer
Refolo, Lorenzo
Project Start
2002-02-01
Project End
2005-01-31
Budget Start
2003-02-01
Budget End
2005-01-31
Support Year
2
Fiscal Year
2003
Total Cost
$195,000
Indirect Cost
Name
State University of New York at Buffalo
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
038633251
City
Buffalo
State
NY
Country
United States
Zip Code
14260
Buzanska, Leonora; Jurga, Marcin; Stachowiak, Ewa K et al. (2006) Neural stem-like cell line derived from a nonhematopoietic population of human umbilical cord blood. Stem Cells Dev 15:391-406
Bharali, Dhruba J; Klejbor, Ilona; Stachowiak, Ewa K et al. (2005) Organically modified silica nanoparticles: a nonviral vector for in vivo gene delivery and expression in the brain. Proc Natl Acad Sci U S A 102:11539-44
Corso, Thomas D; Torres, German; Goulah, Christopher et al. (2005) Transfection of tyrosine kinase deleted FGF receptor-1 into rat brain substantia nigra reduces the number of tyrosine hydroxylase expressing neurons and decreases concentration levels of striatal dopamine. Brain Res Mol Brain Res 139:361-6
Corso, Thomas D; Torres, German; Goulah, Christopher et al. (2005) Assessment of viral and non-viral gene transfer into adult rat brains using HSV-1, calcium phosphate and PEI-based methods. Folia Morphol (Warsz) 64:130-44
Hu, Yafang; Fang, Xiaohong; Dunham, Star M et al. (2004) 90-kDa ribosomal S6 kinase is a direct target for the nuclear fibroblast growth factor receptor 1 (FGFR1): role in FGFR1 signaling. J Biol Chem 279:29325-35
Stachowiak, E K; Fang, X; Myers, J et al. (2003) cAMP-induced differentiation of human neuronal progenitor cells is mediated by nuclear fibroblast growth factor receptor-1 (FGFR1). J Neurochem 84:1296-312
Stachowiak, Michal K; Fang, Xiaohong; Myers, Jason M et al. (2003) Integrative nuclear FGFR1 signaling (INFS) as a part of a universal ""feed-forward-and-gate"" signaling module that controls cell growth and differentiation. J Cell Biochem 90:662-91
Somanathan, Suryanarayan; Stachowiak, Ewa K; Siegel, Alan J et al. (2003) Nuclear matrix bound fibroblast growth factor receptor is associated with splicing factor rich and transcriptionally active nuclear speckles. J Cell Biochem 90:856-69
Myers, Jason M; Martins, Gabriel G; Ostrowski, Jacek et al. (2003) Nuclear trafficking of FGFR1: a role for the transmembrane domain. J Cell Biochem 88:1273-91