|Title of Center Grant:
||Yale/NIDA Neuroproteomics Center
||2 P30 DA018343
|Total Budget Period:
||8/23/2004 – 5/31/2020
|Theme of Center:
||"Proteomics of Altered Signaling in Addiction"
Angus C. Nairn
Co-Director, Yale/NIDA Neuroproteomics Center
Charles B. Murphy Professor of Psychiatry and Pharmacology
Yale University School of Medicine
Kenneth R. Williams
Professor (Adj.) Research
Dept. of Molecular Biophysics & Biochemistry
Yale University School of Medicine
||National Institute on Drug Abuse (NIDA)
National Institutes of Health (NIH)
- Bioinformatics and Biostatistics [BBC, includes High Performance Computing and Yale Protein Expression Database (YPED)]
- Discovery Proteomics (DPC)
- Targeted Proteomics (TPC)
- Pilot Research Projects
||23 investigators from 10 institutions including Indiana University, Mount Sinai School of Medicine, Rockefeller University, University of Chicago, University of Connecticut Health Center, University of Pittsburgh, University of Texas Southwestern Medical Center, Tufts University, and Yale University
- Bring together faculty working at the forefront of such key neuroscience areas as signal transduction, plasticity, neuronal morphogenesis, lipid metabolism in neuronal signaling and synaptic function, and response to psychotropic drugs to work in unique synergy with experts in the Keck Laboratory in MS/proteomics, biophysics, biostatistics, bioinformatics, and high performance computing.
- Use cutting edge proteomic technologies like isobaric tags for relative and absolute quantitation (iTRAQ), LC/MS Label Free Quantitation (LFQ), stable isotope labeling by amino acids in cell culture (SILAC), multi-dimensional protein identification technology (MudPIT), phosphoproteome enrichment and profiling, and targeted quantitation of pre-selected proteins and their post-translational modifications using multiple reaction monitoring (MRM) and Parallel Reaction Monitoring (PRM) to analyze neuronal signal transduction mechanisms and the adaptive changes in these processes that occur in intracellular signaling pathways in brain reward areas in response to drugs of abuse including cannabis, cocaine, nicotine, and opioids.
- Very positively leverage the Center’s proteome analyses by coupling them with the biophysical analyses needed to understand protein function and the phosphoinositide and other lipid analyses needed to provide an increasingly biological “system-wide” view of the impact of drug abuse.
- To address the huge level of cellular and sub-cellular heterogeneity in the central nervous system, which is the greatest obstacle standing in the way of progress towards understanding the adaptive molecular changes underlying addictive drug action, we will launch a multi-pronged initiative that will use laser capture, fluorescence cytometry-related, and immuno-affinity technologies in conjunction with transgenic and viral methods to analyze the proteomes of the single types of neurons that define the circuits that underlie the actions and addictive properties of drugs of abuse.
- To identify the multiple PTMs that occur in individual proteoforms, for example, the combinatorial epigenetic changes in histone modification that are a major interest of several Center investigators, the DPC will implement “Top-Down” analyses.
- To improve peptide identification rates the DPC will collaborate with the BBC to create a completely novel resource that will deeply integrate RNA and protein-level analyses at isoform resolution to enable MS/MS database searches to be carried out on brain region- and cell type-specific proteomes that have been predicted based on RNA-sequencing analyses.
- Provide training and constantly strive to improve existing and develop new proteomics technologies that can be applied to biological questions related to the actions of drugs of abuse.
- Use the Pilot Research Project Core as a cornerstone in the Center’s efforts to 1) encourage strong mentoring relationships that will help attract and train future outstanding scientists in the field of substance abuse and 2) help enable investigators to acquire the preliminary data needed to bring novel and highly promising research ideas relevant to the Center's theme to the point where they can successfully compete for NIH and other grant support.
||To substantially increase understanding of the biochemical mechanisms that underlie substance abuse and its treatment.