{
"METABOLOMICS WORKBENCH":{"VERSION":"1","CREATED_ON":"November 17, 2016, 3:33 pm"},

"PROJECT":{"PROJECT_TITLE":"The alpha-1A adrenergic receptor agonist A61603 reduces cardiac polyunsaturated fatty acid","PROJECT_TYPE":"GC-MS non targeted qualitative analysis","PROJECT_SUMMARY":"Introduction Alpha-1-adrenergic receptors (α1-ARs) are G-protein coupled receptors (GPCRs) with three highly homologous subtypes (α1A, α1B, and α1D). Of these three subtypes, only the α1A and α1B are expressed in the heart. Multiple pre-clinical models of heart injury demonstrate cardioprotective roles for the α1A. Non-selective α1-AR activation promotes glycolysis in the heart, but the functional α1-AR subtype and broader metabolic effects have not been studied. Objectives Given the high metabolic demands of the heart and previous evidence indicating benefit from α1A activation, we chose to investigate the effects of α1A activation on the cardiac metabolome in vivo. Methods Mice were treated for 1 week with a low, subpressor dose of A61603, a highly selective and potent α1A agonist. Cardiac tissue and serum were analyzed using a non-targeted metabolomics approach. Results We identified previously unrecognized metabolic responses to α1A activation,most notably broad reduction in the abundance of polyunsaturated fatty acids(PUFAs) and endocannabinoids (ECs). Conclusion Given the well characterized roles of PUFAs and ECs in inflammatory pathways, these findings suggest a possible role for cardiac α1A-ARs in the regulation of inflammation and may offer novel insight into the mechanisms underlying the cardioprotective benefit of selective pharmacologic α1A activation.","INSTITUTE":"University of North Carolina","DEPARTMENT":"McAllister heart Institute, Department of Internal medicine","LABORATORY":"Multiple Centers","LAST_NAME":"Ilaiwy, Willis","FIRST_NAME":"Amro, Monte","ADDRESS":"111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA","EMAIL":"amroilaiwy@gmail.com, monte_willis@med.unc.edu","PHONE":"210-596-0171","FUNDING_SOURCE":"NIH, Fondation Leducq"},

"STUDY":{"STUDY_TITLE":"The alpha-1A adrenergic receptor agonist A61603 reduces cardiac polyunsaturated fatty acid-Heart raw data","STUDY_TYPE":"GC-MS non-targeted metabolomic profiling","STUDY_SUMMARY":"Studies of skeletal muscle disuse either in patients on bed rest or experimentally in animals(immobilization) have demonstrated that decreased protein synthesis is common, with transient parallel increases in protein degradation. Muscle disuse atrophy involves a process of transition from slow to fast myosin fiber types 6 . A shift toward glycolysis, decreased capacity for fat oxidation, and substrate accumulation in atrophied muscles have been reported as has accommodation of the liver with an increased gluconeogenic capacity. Recent studies have modeled skeletal muscle disuse by using cyclic stretch of differentiated myotubes (C2C12), which mimics the loading pattern of mature skeletal muscle, followed by cessation of stretch.We utilized this model to determine the metabolic changes using non-targeted metabolomics analysis of the media. We identified increases in amino acids resulting from protein degradation (largely sarcomere) that occurs with muscle atrophy that are involved in feeding the Kreb’s cycle through anaplerosis. Specifically, we identified increased alanine/proline metabolism (significantly elevated proline, alanine, glutamine, and asparagine) and increased -ketoglutaric acid, the proposed Kreb’s cycle intermediate being fed by the alanine/proline metabolic anaplerotic mechanism. Additionally, several unique pathways not clearly delineated in previous studies of muscle unloading were seen, including: 1) elevated ethanolamine and elevated keto-acids (e.g. 2-ketoleucine and 2-keovaline) represent intermediates in the Ehlrich amino acid degradation pathway, which feeds into a metabolic pathway supplying acetyl-CoA and 2-hydroxybutyrate (also significantly increased); and 2) elevated guanine, an intermediate of purine metabolism, was seen at 12 hours unloading. Given the interest in targeting different aspects of the ubiquitin proteasome system to inhibit protein degradation, this C2C12 system may allow the identification of direct and indirect alterations in metabolism due to anaplerosis or through other yet to be identified mechanisms using a non-targeted metabolomics approach.","INSTITUTE":"University of North Carolina","DEPARTMENT":"McAllister Heart Institute","LABORATORY":"Mutliple Centers","LAST_NAME":"Ilaiwy, WIllis","FIRST_NAME":"Amro, Monte","ADDRESS":"111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA","EMAIL":"amroilaiwy@gmail.com, monte_willis@med.unc.edu","PHONE":"919-3607599"},

"SUBJECT":{"SUBJECT_TYPE":"Animal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"UCSF-01005",
"Factors":{"Treatment":"Vehicle"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01006",
"Factors":{"Treatment":"Vehicle"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01009",
"Factors":{"Treatment":"Vehicle"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01010",
"Factors":{"Treatment":"Vehicle"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01013",
"Factors":{"Treatment":"Vehicle"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01014",
"Factors":{"Treatment":"Vehicle"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01017",
"Factors":{"Treatment":"Vehicle"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01018",
"Factors":{"Treatment":"Vehicle"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01003",
"Factors":{"Treatment":"A61603"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01004",
"Factors":{"Treatment":"A61603"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01007",
"Factors":{"Treatment":"A61603"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01008",
"Factors":{"Treatment":"A61603"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01011",
"Factors":{"Treatment":"A61603"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01012",
"Factors":{"Treatment":"A61603"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01015",
"Factors":{"Treatment":"A61603"}
},
{
"Subject ID":"-",
"Sample ID":"UCSF-01016",
"Factors":{"Treatment":"A61603"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Mice were sacrificed under deep isoflurane anesthesia, blood was collected by cardiac puncture, clotted, and 100–200 μL serum was snap frozen in liquid nitrogen. The heart was dissected, the atria and great vessels were excised,and the ventricles were allowed to beat in cold PBS to clear blood."},

"TREATMENT":{"TREATMENT_SUMMARY":"N/A"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"sample extract reconstituted in solvents containing a series of standards at fixed concentrations to ensure injection and chromatographic consistency. One aliquot was analyzed using acidic positive ion conditions, chromatographically optimized for more hydrophilic compounds. The extract was gradient eluted from a C18 column (Waters UPLC BEH C18-2.1 × 100 mm, 1.7 μm) using water and methanol, containing 0.05 % perfluoropentanoic acid (PFPA) and 0.1 % formic acid (FA). Another aliquot was also analyzed using acidic positive ion conditions,however it was chromatographically optimized for more hydrophobic compounds. In this method, the extract was gradient eluted from the same C18 column using methanol, acetonitrile, water, 0.05 % PFPA and 0.01 % FA and was operated at an overall higher organic content. Another aliquot was analyzed using basic negative ion optimized conditions using a separate dedicated C18 column. The basic extracts were gradient eluted from the column using methanol and water"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Waters Acquity","COLUMN_NAME":"Waters Acquity BEH C18 (100 x 2mm, 1.7um)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Agilent 5975","INSTRUMENT_TYPE":"Single quadrupole","MS_TYPE":"EI","ION_MODE":"POSITIVE"},

"MS_METABOLITE_DATA":{
"Units":"Peak values (scaled)",

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}