Discovery of neuroproteomic determinants of pain transmission using differential and systematic proteome exploration strategies

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The treatment of pain perception following traumatized nervous tissue leading to hyperalgesia and allodynia in neuropathic animals has been widely investigated with limited success. This research program will be initiated in three phases. The initial phase will include differential proteome exploration in the central nervous system using preprotachykinins (PPT-A) knockout and wild type mice for the identification of specific proteins encoded by the preprotachykinins gene with an emphasis on proneuropeptides of tachykinins neuropeptides. The second phase will include a systematic proteome exploration in the central nervous system including the identification, characterization and quantification of the neuroproteome with a special interest on proneuropeptides of tachykinins neuropeptides in a validated rat model of neuropathic pain. We developed an expertise in the analysis of peptides and proteins from spinal cords using, specific extraction/concentration techniques, digestion methods, SDS-PAGE, 2D gel electropheresis, capillary HPLC, ion trap mass spectrometry and MALDI-TOF. Protein quantification will be performed using multiplexed isobaric tagging techniques for biomarker discovery to characterize protein expression ratio profiles, performed principal component analysis (PCA) and define a multiparametric profile with a primary focus in preprotachykinins gene related proteins for diagnostic and prognostic purposes. Moreover, the third objective will include the evaluation of the neuroproteome regulation following the administration of agonist/antagonist of Transient Receptor Potential Channel Vanilloid 1 (TRPV1). TRPV1 receptors are involved in the transmission and modulation of pain and TRPV1 ligands shown promising results to alleviate pain. The general objective of this proposal is to understand the mechanism and the condition from which neuroproteins with a special interest to preprotachykinins gene related proteins are regulated and foster better mechanistic knowledge to support the development of new medicines to treat neuropathic and chronic pain.