18. GOALSFORFELLOWSHTIPRAININGANDCAREER While I have formal training in genetics (B.S.) and some practical molecular biology experience, I have not had the opportunity, as of yet, to utilize these techniques to investigate a physiological process, such as neurodegeneration. The fellowship will give me the training and opportunity to develop a well-defined research plan in the field of cell-death and neurodegeneration. Since a significant portion of my research proposal utilizes molecular biology techniques, completion of the proposed research will enable me to learn the conceptual, theoretical and practical application of these research skills to elucidate critical pathways in neurodegenerative diseases. Molecular biology techniques are often utilized in this area of research and therefore my training will not only enable me to complete my proposed postdoctoral research project but will help me become an independent investigator working in the field of neurodegeneration and aging. Dr. Lisa Ellerby's laboratory is ideal for this training since Dr. Ellerby has years of experience applying molecular biology techniques to understand oxidative injury, activation of apoptotic pathways and the pathogenesis of polyglutamine diseases. SPONSOR 19. NAME AND DEGREE(S) Lisa M. Ellerby, Ph.D. 20. POSITION/RANK Assistant Professor 21. RESEARCIHNTERESTS/AREAS Aging, Apoptosis and Neurodegenerative Diseases RESEARCH PROPOSAL 22. DESCRIPTION (Do not exceed space provided) Huntington's disease (HD) is an autosomal-dominant progressive neurodegenerative disorder, which results from a CAG expansion in exon 1 of the huntingtin gene (htt). Currently it is unknown how the mutant huntingtin protein (Htt) promotes the selective neurodegeneration of HD. Multiple lines of evidence support the hypothesis that production of small N-terminal Htt fragments plays a key role in the pathogenesis of the disease. Preliminary evidence in our laboratory shows that calpain cleaves Htt at multiple sites to produce small N- terminal fragments and that the active form of calpain is substantially increased in the caudate of HD patients. In order to address the hypothesis that production of calpain-derived Htt fragments contributes to the specific neurodegeneration observed in HD, the predicted calpain cleavage sites in normal and expanded htt will be mutated to see if changes in production of calpain-dedved Htt fragments influence cellular viability. In addition normal and expanded calpain-derived Htt fragments will be produced and expressed in striatal neurons to correlate the cytotoxicity of the fragments with nuclear migration and aggregation. Immunoprecipitation with a Htt antibody will also be utilized to determine whether the normal and/or expanded form of Htt physically associates with calpain to produce calpain-derived Htt fragments. Lastly, the cell types in the caudate of HD displaying changes in sub-cellular distribution of calpain will be identified and it will be determined if calpain co- localizes with Htt through double labeling of striatal neurons overexpressing calpain-derived Htt fragments. F_HS416-1 (Rev. 12/98) Form Page 2 BE cc NAME(Last,first,middleinitial) Individual NRSA Application Table of Contents ========================================Section End===========================================
| Schilling, Birgit; Gafni, Juliette; Torcassi, Cameron et al. (2006) Huntingtin phosphorylation sites mapped by mass spectrometry. Modulation of cleavage and toxicity. J Biol Chem 281:23686-97 |
| Jin, Kunlin; LaFevre-Bernt, Michelle; Sun, Yunjuan et al. (2005) FGF-2 promotes neurogenesis and neuroprotection and prolongs survival in a transgenic mouse model of Huntington's disease. Proc Natl Acad Sci U S A 102:18189-94 |
| Hermel, E; Gafni, J; Propp, S S et al. (2004) Specific caspase interactions and amplification are involved in selective neuronal vulnerability in Huntington's disease. Cell Death Differ 11:424-38 |
| Gafni, Juliette; Hermel, Evan; Young, Jessica E et al. (2004) Inhibition of calpain cleavage of huntingtin reduces toxicity: accumulation of calpain/caspase fragments in the nucleus. J Biol Chem 279:20211-20 |
