#METABOLOMICS WORKBENCH STUDY_ID:ST000185 ANALYSIS_ID:AN000284 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 AQM020 CH:METHODS_FILENAME QTOF-002-HILIC-35min-1mm-No_Insert.m.zip CH:INSTRUMENT_NAME Agilent 1260 CH:COLUMN_NAME Phenomenex Luna NH2 (150 x 1mm, 3um) CH:CHROMATOGRAPHY_TYPE HILIC #ANALYSIS AN:LABORATORY_NAME MRC2 (University of Michigan) AN:ANALYSIS_TYPE MS AN:ACQUISITION_PARAMETERS_FILE QTOF-002-HILIC-35min-1mm-No_Insert.m.zip #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 6530 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE #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 2 or 3-phosphoglycerate_2PG/3PG 0.8978 0.3447 0.2680 6.4525 2.9002 6.3058 6-Phosphogluconate 0.6816 0.5188 0.3125 1.9113 1.4771 3.7586 Acetyl-CoA 0.1221 0.7683 0.5710 0.8624 ADP 21.4299 16.3787 15.1622 67.1431 40.6175 51.8677 AMP 9.3438 7.0078 7.4143 178.2130 68.4896 122.5572 ATP 66.8491 45.4450 39.1587 81.5229 90.1356 68.5431 Citrate or Isocitrate 30.3007 77.3516 33.7218 208.7459 129.0972 71.8109 flavin adenine dinucleotide 0.0975 0.1405 0.1166 0.3954 0.2555 0.3425 Fructose 1_6-bisphosphate 3.4687 1.1181 1.1003 16.3895 9.1119 15.0493 Glucose 6-phosphate or fructose-6-phosphate 16.2806 11.1167 7.8941 137.8086 33.0055 107.1428 Malate 42.5975 95.4928 36.2395 44.2528 36.1339 nicotinamide adenine dinucleotide phosphate oxidized 1.2664 1.1893 1.0154 3.6640 2.3189 3.4695 Phosphoenolpyruvate 1.7034 3.3100 0.9094 6.3348 7.7797 4.6110 Ribose-5-phosphate or xylulose 5-phosphate 4.0956 1.1797 1.5413 11.6761 4.3669 8.9582 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 2 or 3-phosphoglycerate_2PG/3PG 724 C00597 2PG/3PG UM_Target_ID 6-Phosphogluconate 91493 C00345 6PG UM_Target_ID Acetyl-CoA 444493 C00024 aCoA UM_Target_ID ADP 6022 C00008 ADP UM_Target_ID AMP 6083 C00020 AMP UM_Target_ID ATP 5957 C00002 ATP UM_Target_ID Citrate or Isocitrate 311 C00158 CIT/ICIT UM_Target_ID flavin adenine dinucleotide 643975 C00016 FAD UM_Target_ID Fructose 1,6-bisphosphate 172313 C00354 FBP UM_Target_ID Glucose 6-phosphate or fructose-6-phosphate 5958 C00092 G6P/F6P UM_Target_ID Malate 222656 C00149 MAL UM_Target_ID nicotinamide adenine dinucleotide phosphate oxidized 5886 C00006 NADP UM_Target_ID Phosphoenolpyruvate 1005 C00074 PEP UM_Target_ID Ribose-5-phosphate or xylulose 5-phosphate 77982 C00117 R5P/X5P UM_Target_ID METABOLITES_END #END