#METABOLOMICS WORKBENCH STUDY_ID:ST000185 ANALYSIS_ID:AN000285 PROJECT_ID:PR000158 VERSION 1 CREATED_ON 01-15-2019 #PROJECT PR:PROJECT_TITLE LEC metabolomics PR:PROJECT_SUMMARY Comparison of metabolomic profile in LECs derived from normal and shunt animals PR:INSTITUTE University of California, San Francisco PR:DEPARTMENT Pediatrics PR:LABORATORY Fineman Lab PR:LAST_NAME Fineman PR:FIRST_NAME Jeffrey PR:ADDRESS San Francisco, CA PR:EMAIL jeff.fineman@ucsf.edu PR:PHONE 415-502-6390 PR:DOI http://dx.doi.org/10.21228/M8T01Q #STUDY ST:STUDY_TITLE Fetal Lambs vascular graft Normal v Shunt LECs ST:STUDY_TYPE Glycolysis/TCA/Nucleotide analysis (tissue/cells) ST:STUDY_SUMMARY We recently reported superior right ventricle (RV) performance in response to ST:STUDY_SUMMARY acute afterload challenge in lambs with a model of congenital heart disease with ST:STUDY_SUMMARY chronic left-to-right cardiac shunts. Compared with control animals, shunt lambs ST:STUDY_SUMMARY demonstrated increased contractility because of an enhanced Anrep effect (the ST:STUDY_SUMMARY slow increase in contractility following myocyte stretch). This advantageous ST:STUDY_SUMMARY physiological response may reflect preservation of a fetal phenotype, since the ST:STUDY_SUMMARY RV of shunt lambs remains exposed to increased pressure postnatally. Nitric ST:STUDY_SUMMARY oxide (NO) production by NO synthase (NOS) is activated by myocyte stretch and ST:STUDY_SUMMARY is a necessary intermediary of the Anrep response. The purpose of this study was ST:STUDY_SUMMARY to test the hypothesis that NO signaling is increased in the RV of fetal lambs ST:STUDY_SUMMARY compared with controls and shunt lambs have persistence of this fetal pattern. ST:STUDY_SUMMARY An 8-mm graft was placed between the pulmonary artery and aorta in fetal lambs ST:STUDY_SUMMARY (shunt). NOS isoform expression, activity, and association with activating ST:STUDY_SUMMARY cofactors were determined in fetal tissue obtained during late-gestation and in ST:STUDY_SUMMARY 4-wk-old juvenile shunt and control lambs. We demonstrated increased RNA and ST:STUDY_SUMMARY protein expression of NOS isoforms and increased total NOS activity in the RV of ST:STUDY_SUMMARY both shunt and fetal lambs compared with control. We also found increased NOS ST:STUDY_SUMMARY activation and association with cofactors in shunt and fetal RV compared with ST:STUDY_SUMMARY control. These data demonstrate preserved fetal NOS phenotype and NO signaling ST:STUDY_SUMMARY in shunt RV, which may partially explain the mechanism underlying the adaptive ST:STUDY_SUMMARY response to increased afterload seen in the RV of shunt lambs. Research is ST:STUDY_SUMMARY published, core data not used but project description is relevant: ST:STUDY_SUMMARY http://ajpheart.physiology.org/content/309/1/H157.long ST:INSTITUTE University of Michigan ST:DEPARTMENT Biomedical Research Core Facilities ST:LABORATORY Metabolomics core ST:LAST_NAME Kachman ST:FIRST_NAME Maureen ST:ADDRESS 6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714 ST:EMAIL mkachman@umich.edu ST:PHONE - #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Ovis aries SU:TAXONOMY_ID 9940 SU:SPECIES_GROUP Mammal #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS SU0011546 S00017020 Type:increased flow SUBJECT_SAMPLE_FACTORS SU0011547 S00017021 Type:increased flow SUBJECT_SAMPLE_FACTORS SU0011548 S00017022 Type:increased flow SUBJECT_SAMPLE_FACTORS SU0011543 S00017017 Type:normal control SUBJECT_SAMPLE_FACTORS SU0011544 S00017018 Type:normal control SUBJECT_SAMPLE_FACTORS SU0011545 S00017019 Type:normal control #COLLECTION CO:SAMPLE_TYPE Endothelial Cells CO:COLLECTION_SUMMARY - #TREATMENT TR:TREATMENT_SUMMARY - #SAMPLEPREP SP:SAMPLEPREP_PROTOCOL_FILENAME Gly-TCA-nucleotides_analysis_protocol-2015-03-09.docx SP:SAMPLEPREP_SUMMARY - #CHROMATOGRAPHY CH:METHODS_ID AQM011 CH:METHODS_FILENAME ALPHA_KETO_ACIDS-FULL.M.zip CH:INSTRUMENT_NAME Agilent 7890A CH:COLUMN_NAME Agilent DB5-MS (30m × 0.25mm, 0.25um) CH:CHROMATOGRAPHY_TYPE GC #ANALYSIS AN:LABORATORY_NAME MRC2 (University of Michigan) AN:ANALYSIS_TYPE MS AN:ACQUISITION_PARAMETERS_FILE ALPHA_KETO_ACIDS-FULL.M.zip #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 5975C MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS pmol/µg protein MS_METABOLITE_DATA_START Samples S00017020 S00017021 S00017022 S00017017 S00017018 S00017019 Factors Type:increased flow Type:increased flow Type:increased flow Type:normal control Type:normal control Type:normal control alpha-Ketoglutarate 23.9633 97.6063 25.9218 10.2784 18.6104 4.0044 Aspartate 28.9800 53.3015 30.6237 46.9567 68.9472 14.6040 fumarate 7.9338 15.4468 6.3243 9.4925 16.9246 3.5431 Glutamate 790.1819 2101.6621 615.9909 600.9693 920.9131 182.8471 Lactate 16463.0497 90098.3522 25552.9804 2325.6219 5968.9836 684.1921 pyruvate 48.1264 173.4467 68.0630 44.0311 115.6702 13.8828 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant alpha-Ketoglutarate 51 C00026 AKG UM_Target_ID Aspartate 5960 C00049 ASP UM_Target_ID fumarate 444972 C00122 FUM UM_Target_ID Glutamate 611 GLU UM_Target_ID Lactate 107689 C00186 LAC UM_Target_ID pyruvate 1060 C00022 PYR UM_Target_ID METABOLITES_END #END