Parkinson disease (PD) is a debilitating neurodegenerative disease for which there are no effective neuroprotective therapies. Converging data from post-mortem brain studies, oxidative toxin models, and PD genetics all implicate altered mitochondrial kinase networks in PD pathogenesis. Mutations in the mitochondrial PTEN-induced kinase 1 (PINK1) occur in familial and possibly sporadic PD. The candidate is an established and productive investigator in the signaling regulation of neuronal fate in culture and animal models of PD pathogenesis. The candidate's published and preliminary work implicate mitochondrial targeting of extracellular signal regulated protein kinase 2 (ERK2) and decreased PINK1 signaling in promoting dopaminergic neuronal cell death. Overexpression of wild type PINK1 confers neuroprotection in cell death models. Thus, identifying downstream targets of PINK1, and of mitochondrial targets of redox-activated ERK2, would represent important steps in understanding mechanisms that regulate neuronal survival and death. Identification of substrates for novel kinases such as PINK1, and of context specific ERK2 targets, can be accomplished using the powerful techniques of phospho-proteomics and mass spectroscopy. This proposal capitalizes on the strong cell signaling, proteomic/metabolomic, pharmaceutical, and neurodegeneration research environments at the University of Pittsburgh. The candidate's immediate goals are to obtain training in mass spectrometry and phospho-proteomics under the mentorship of Billy Day, Director of the Pittsburgh Proteomic Core Facility. Co-mentor Bruce Freeman, expert in free radical biochemistry, will provide complementary training in basic redox proteomics. In addition to short courses, seminars, and technical workshops, practical experience will be derived from a pilot project involving ATP-binding pocket mutagenesis and mass spectrometry to identify substrates of PINK1. Lay summary: This career enhancement award will provide an established physician-scientist studying neurodegenerative diseases with essential cross-training in highly specialized target identification technologies. Solidifying interdisciplinary collaborations will enable the principal investigator to more rapidly translate knowledge on factors that determine whether or not neurons can successfully adapt to disease promoting stresses into novel targets for designing neuroprotective PD therapies. ? ? ? ? ?
