#METABOLOMICS WORKBENCH krjonscher_20220420_084738 DATATRACK_ID:3208 STUDY_ID:ST002192 ANALYSIS_ID:AN003588 PROJECT_ID:PR001389 VERSION 1 CREATED_ON June 2, 2022, 7:25 am #PROJECT PR:PROJECT_TITLE Protective effects of maternal PQQ on hepatic lipid metabolism throughout the PR:PROJECT_TITLE lifespan: Adult Study PR:PROJECT_TYPE Diet study and fetal programming PR:PROJECT_SUMMARY Maternal obesity and consumption of a high-fat diet significantly elevate risk PR:PROJECT_SUMMARY for pediatric non-alcoholic fatty liver disease (NAFLD), affecting 10% of PR:PROJECT_SUMMARY children in the US. Almost half of these children are diagnosed with PR:PROJECT_SUMMARY nonalcoholic steatohepatitis (NASH), a leading etiology for liver transplant. PR:PROJECT_SUMMARY Animal models show that signs of liver injury and perturbed lipid metabolism PR:PROJECT_SUMMARY asso-ciated with NAFLD begin in utero; however, safe dietary therapeutics to PR:PROJECT_SUMMARY blunt developmental programming of NAFLD are unavailable. Using a mouse model of PR:PROJECT_SUMMARY maternal Western-style diet (WD), we previously showed that pyrroloquinoline PR:PROJECT_SUMMARY quinone (PQQ), a potent dietary antioxidant, protected offspring of WD-fed dams PR:PROJECT_SUMMARY from development of NAFLD and NASH. Here, we used untargeted mass PR:PROJECT_SUMMARY spectrometry-based lipidomics to delineate lipotoxic effects of WD on offspring PR:PROJECT_SUMMARY liver and identify lipid targets of PQQ. PQQ exposure during pregnancy altered PR:PROJECT_SUMMARY hepatic lipid profiles of WD-exposed offspring, upregulating peroxisome PR:PROJECT_SUMMARY proliferator-activated receptor (PPAR) α signaling and mitochondrial fatty acid PR:PROJECT_SUMMARY oxidation to markedly attenuate triglyceride accumulation beginning in utero. PR:PROJECT_SUMMARY Surprisingly, the abundance of very long-chain ceramides, important in promoting PR:PROJECT_SUMMARY gut barrier and hepatic function, was significantly elevated in PQQ-treated PR:PROJECT_SUMMARY offspring. PQQ exposure reduced the hepatic PR:PROJECT_SUMMARY phosphatidylcho-line/phosphatidylethanolamine (PC/PE) ratio in WD-fed offspring PR:PROJECT_SUMMARY and improved glucose toler-ance. Notably, levels of protective n − 3 PR:PROJECT_SUMMARY polyunsaturated fatty acids (PUFAs) were elevated in offspring exposed to PQQ, PR:PROJECT_SUMMARY beginning in utero, and the increase in n − 3 PUFAs persisted into adulthood. PR:PROJECT_SUMMARY Our findings suggest that PQQ supplementation during gestation and lactation PR:PROJECT_SUMMARY augments pathways involved in the biosynthesis of long-chain fatty acids and PR:PROJECT_SUMMARY plays a unique role in modifying specific bioactive lipid species critical for PR:PROJECT_SUMMARY protection against NAFLD risk in later life. PR:INSTITUTE University of Oklahoma Health Sciences Center PR:DEPARTMENT Biochemistry and Molecular Biology, Harold Hamm Diabetes Center PR:LABORATORY Jonscher PR:LAST_NAME Jonscher PR:FIRST_NAME Karen PR:ADDRESS 975 NE 10th Street BRC-N 362A, Oklahoma City, OK, 73104, USA PR:EMAIL karen-jonscher@ouhsc.edu PR:PHONE 3032294620 PR:FUNDING_SOURCE NIDDK #STUDY ST:STUDY_TITLE Amelioration of developmental programming of NAFLD in adult liver using PQQ ST:STUDY_TYPE Pre-natal and Post-natal Diet and PQQ treatment ST:STUDY_SUMMARY Maternal obesity and consumption of a high-fat diet significantly elevate risk ST:STUDY_SUMMARY for pediatric non-alcoholic fatty liver disease (NAFLD), affecting 10% of ST:STUDY_SUMMARY children in the US. Almost half of these children are diagnosed with ST:STUDY_SUMMARY nonalcoholic steatohepatitis (NASH), a leading etiology for liver transplant. ST:STUDY_SUMMARY Animal models show that signs of liver injury and perturbed lipid metabolism ST:STUDY_SUMMARY asso-ciated with NAFLD begin in utero; however, safe dietary therapeutics to ST:STUDY_SUMMARY blunt developmental programming of NAFLD are unavailable. Using a mouse model of ST:STUDY_SUMMARY maternal Western-style diet (WD), we previously showed that pyrroloquinoline ST:STUDY_SUMMARY quinone (PQQ), a potent dietary antioxidant, protected offspring of WD-fed dams ST:STUDY_SUMMARY from development of NAFLD and NASH. Here, we used untargeted mass ST:STUDY_SUMMARY spectrometry-based lipidomics to delineate lipotoxic effects of WD on offspring ST:STUDY_SUMMARY liver and identify lipid targets of PQQ. PQQ exposure during pregnancy altered ST:STUDY_SUMMARY hepatic lipid profiles of WD-exposed offspring, upregulating peroxisome ST:STUDY_SUMMARY proliferator-activated receptor (PPAR) α signaling and mitochondrial fatty acid ST:STUDY_SUMMARY oxidation to markedly attenuate triglyceride accumulation beginning in utero. ST:STUDY_SUMMARY Surprisingly, the abundance of very long-chain ceramides, important in promoting ST:STUDY_SUMMARY gut barrier and hepatic function, was significantly elevated in PQQ-treated ST:STUDY_SUMMARY offspring. PQQ exposure reduced the hepatic ST:STUDY_SUMMARY phosphatidylcho-line/phosphatidylethanolamine (PC/PE) ratio in WD-fed offspring ST:STUDY_SUMMARY and improved glucose toler-ance. Notably, levels of protective n − 3 ST:STUDY_SUMMARY polyunsaturated fatty acids (PUFAs) were elevated in offspring exposed to PQQ, ST:STUDY_SUMMARY beginning in utero, and the increase in n − 3 PUFAs persisted into adulthood. ST:STUDY_SUMMARY Our findings suggest that PQQ supplementation during gestation and lactation ST:STUDY_SUMMARY augments pathways involved in the biosynthesis of long-chain fatty acids and ST:STUDY_SUMMARY plays a unique role in modifying specific bioactive lipid species critical for ST:STUDY_SUMMARY protection against NAFLD risk in later life. ST:INSTITUTE University of Oklahoma Health Sciences Center ST:DEPARTMENT Biochemistry and Molecular Biology, Harold Hamm Diabetes Center ST:LABORATORY Jonscher ST:LAST_NAME Jonscher ST:FIRST_NAME Karen ST:ADDRESS 975 NE 10th Street BRC-N 362A, Oklahoma City, OK, 73104, USA ST:EMAIL karen-jonscher@ouhsc.edu ST:PHONE 3032294620 ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 24 ST:NUM_MALES 24 ST:PUBLICATIONS Jonscher, et al FASEB J 2017; Friedman, et al Hepatol Commun 2018 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:AGE_OR_AGE_RANGE 20-24 weeks SU:GENDER Male SU:ANIMAL_HOUSING Vivarium University of Colorado Anschutz Medical Campus SU:ANIMAL_LIGHT_CYCLE 12/12 SU:ANIMAL_FEED CH; 2019; Envigo, Indianapolis, IN or .D TD.88137; Envigo SU:ANIMAL_WATER Water or treated with 1.25 mg/L PQQ #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - 1991A Diet:CTL SUBJECT_SAMPLE_FACTORS - 1991B Diet:CTL SUBJECT_SAMPLE_FACTORS - 1991C Diet:CTL SUBJECT_SAMPLE_FACTORS - 1996A Diet:CTL SUBJECT_SAMPLE_FACTORS - 1996B Diet:CTL SUBJECT_SAMPLE_FACTORS - 1996C Diet:CTL SUBJECT_SAMPLE_FACTORS - 1990A Diet:CTL SUBJECT_SAMPLE_FACTORS - 1990B Diet:CTL SUBJECT_SAMPLE_FACTORS - 1990C Diet:CTL SUBJECT_SAMPLE_FACTORS - 5329C Diet:CTL SUBJECT_SAMPLE_FACTORS - 5329A Diet:CTL SUBJECT_SAMPLE_FACTORS - 5329B Diet:CTL SUBJECT_SAMPLE_FACTORS - 389A Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 389B Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 389C Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 330A Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 330B Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 330C Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 358A Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 358B Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 358C Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 333A Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 333B Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 333C Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 362A Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 362B Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 362C Diet:CTL PQQ SUBJECT_SAMPLE_FACTORS - 754A Diet:WD SUBJECT_SAMPLE_FACTORS - 754B Diet:WD SUBJECT_SAMPLE_FACTORS - 754C Diet:WD SUBJECT_SAMPLE_FACTORS - 1007A Diet:WD SUBJECT_SAMPLE_FACTORS - 1007B Diet:WD SUBJECT_SAMPLE_FACTORS - 1007C Diet:WD SUBJECT_SAMPLE_FACTORS - 5328B Diet:WD SUBJECT_SAMPLE_FACTORS - 5328C Diet:WD SUBJECT_SAMPLE_FACTORS - 5328A Diet:WD SUBJECT_SAMPLE_FACTORS - 372A Diet:WD SUBJECT_SAMPLE_FACTORS - 372B Diet:WD SUBJECT_SAMPLE_FACTORS - 372C Diet:WD SUBJECT_SAMPLE_FACTORS - 386A Diet:WD SUBJECT_SAMPLE_FACTORS - 386B Diet:WD SUBJECT_SAMPLE_FACTORS - 386C Diet:WD SUBJECT_SAMPLE_FACTORS - 361A Diet:WD SUBJECT_SAMPLE_FACTORS - 361B Diet:WD SUBJECT_SAMPLE_FACTORS - 361C Diet:WD SUBJECT_SAMPLE_FACTORS - 989C Diet:WD SUBJECT_SAMPLE_FACTORS - 989A Diet:WD SUBJECT_SAMPLE_FACTORS - 989B Diet:WD SUBJECT_SAMPLE_FACTORS - 773A Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 773B Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 773C Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 768A Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 768B Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 768C Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 901A Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 901B Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 901C Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 767A Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 767B Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 767C Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 703A Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 703B Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 703C Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 756A Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 756B Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 756C Diet:WD PQQ SUBJECT_SAMPLE_FACTORS - 179A Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 179B Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 179C Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 181A Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 181B Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 181C Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 743A Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 743B Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 743C Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 739A Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 739B Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 739C Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 727A Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 727B Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 727C Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 766A Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 766B Diet:WD PQQ/WD SUBJECT_SAMPLE_FACTORS - 766C Diet:WD PQQ/WD #COLLECTION CO:COLLECTION_SUMMARY Livers were excised and snap frozen then 10-15 mg aliquots sent to the West CO:COLLECTION_SUMMARY Coast Metabolomics Center for analysis following standard protocols. CO:SAMPLE_TYPE Liver CO:STORAGE_CONDITIONS -80℃ CO:COLLECTION_VIALS Cryotubes CO:STORAGE_VIALS Cryotubes #TREATMENT TR:TREATMENT_SUMMARY Dams and offspring were fed either chow (CH) or western-style diet (WD), with or TR:TREATMENT_SUMMARY without PQQ in drinking water. A subset of WD-exposed offspring were weaned onto TR:TREATMENT_SUMMARY WD without PQQ. TR:TREATMENT WD and PQQ TR:TREATMENT_COMPOUND BioPQQ TR:TREATMENT_ROUTE Drinking water, ad libitem TR:TREATMENT_DOSE 1.25 mg/L TR:TREATMENT_DOSEDURATION 17-21 weeks TR:TREATMENT_VEHICLE drinking water #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Tissue was homogenized and lipids extracted following standard protocols at the SP:SAMPLEPREP_SUMMARY WCMC. Samples were prepared in triplicate SP:SAMPLEPREP_PROTOCOL_FILENAME SP_Extraction_Protocol_for_liver_multi-omic.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1200 CH:COLUMN_NAME Waters Acquity CSH C18 (100 x 2.1mm, 1.7um) CH:METHODS_FILENAME SOP_Lipidomic_Analysis_by_UPLC_QTOF.pdf #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE SOP_Lipidomic_Analysis_by_UPLC_QTOF.pdf #MS MS:INSTRUMENT_NAME Agilent 6530 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS See attached file MS:MS_RESULTS_FILE ST002192_AN003588_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END