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Identification of novel signaling regulators of the mu opioid receptor (1066.1)
Author(s) -
Edwards Katie,
LaVigne Justin,
MosesFynn Emmanuel,
Mullen Nate,
Streicher John
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1066.1
Subject(s) - mapk/erk pathway , drug discovery , signal transduction , g protein coupled receptor , μ opioid receptor , opioid , hek 293 cells , microbiology and biotechnology , receptor , small interfering rna , scaffold protein , biology , chemistry , pharmacology , computational biology , rna , bioinformatics , biochemistry , gene
Recent evidence has suggested that specific downstream signaling pathways of the mu opioid receptor (MOR) carry out specific aspects of the behavioral response, such as antinociception or side effects. These findings have led to a drug discovery effort to develop functionally selective MOR ligands which activate the desired cascades (G protein) and not the undesirable cascades (βarrestin2). However, the βarrestin pathway is the only such functionally selective target identified for MOR drug discovery. In order to discover additional novel MOR regulators, we have developed two unbiased screening approaches. First, we have used co‐immunoprecipitation of the drug activated MOR signaling complex in MOR‐expressing CHO cells, followed by proteomic analysis. In addition, we have developed a pooled small‐hairpin RNA screen to find regulators of MOR induced signaling (ERK MAPK readout) in MOR‐expressing HEK293 cells. We present here our current progress using these screens, the outcome of which are lists of candidate MOR signaling regulators. One possible MOR‐regulating scaffold protein has been knocked down in MOR‐expressing CHO cells using small‐interfering RNA and shows evidence of regulating ERK MAPK pathways induced by the MOR. By defining the MOR signaling complex in this way, we hope to identify future targets for functionally selective drug discovery in order to improve the treatment of chronic pain. Grant Funding Source : Supported by UNE COBRE Seed Grant P20GM103643