#METABOLOMICS WORKBENCH rgn5011_20170926_112858 DATATRACK_ID:1246 STUDY_ID:ST000907 ANALYSIS_ID:AN001474 PROJECT_ID:PR000629 VERSION 1 CREATED_ON December 1, 2017, 11:09 am #PROJECT PR:PROJECT_TITLE Murine vitamin A deficiency results in a hypermetabolic state and alterations in PR:PROJECT_TITLE bacterial community structure and metabolism PR:PROJECT_SUMMARY Vitamin A deficiency (A-) is a significant public health problem. To better PR:PROJECT_SUMMARY understand how vitamin A status influences gut microbiota and host metabolism, PR:PROJECT_SUMMARY we systematically analyzed urine, cecum, serum, and liver samples from vitamin A PR:PROJECT_SUMMARY sufficient (A+) and A- mice using 1H NMR-based metabolomics, quantitative PR:PROJECT_SUMMARY (q)PCR, and 16S rRNA gene sequencing coupled with multivariate data analysis. PR:PROJECT_SUMMARY The microbiota in the cecum of A- mice showed compositional as well as PR:PROJECT_SUMMARY functional shifts compared to the microbiota from A+ mice. Targeted 1H NMR PR:PROJECT_SUMMARY analyses revealed significant changes in microbial metabolite concentrations PR:PROJECT_SUMMARY including higher butyrate and hippurate and decreased acetate and PR:PROJECT_SUMMARY 4-hydroxyphenylacetate in A+ relative to A- mice. Bacterial butyrate-producing PR:PROJECT_SUMMARY genes including butyryl-CoA:acetate CoA-transferase and butyrate kinase were PR:PROJECT_SUMMARY significantly higher in bacteria from A+ versus bacteria from A- mice. A - mice PR:PROJECT_SUMMARY had disturbances in multiple metabolic pathways including alterations in energy PR:PROJECT_SUMMARY metabolism (hyperglycemia, glycogenesis, TCA cycle, and lipoprotein PR:PROJECT_SUMMARY biosynthesis) and the A- host showed metabolites indicative of a hypermetabolic PR:PROJECT_SUMMARY state (higher levels of amino acids and nucleic acids). A- mice had PR:PROJECT_SUMMARY hyperglycemia, liver dysfunction, changes in bacterial metabolism, and altered PR:PROJECT_SUMMARY gut microbial communities. Moreover, integrative analyses indicated a strong PR:PROJECT_SUMMARY correlation between gut microbiota and host energy metabolism pathways in the PR:PROJECT_SUMMARY liver. Vitamin A regulates the microbiota, bacterial metabolism and the effects PR:PROJECT_SUMMARY of vitamin A on the microbiota results in alterations to host metabolism. PR:INSTITUTE The Pennsylvania State University (Penn State) PR:LAST_NAME Nichols PR:FIRST_NAME Robert PR:ADDRESS 101 Life science building, University Park, State college, PA, 16803 PR:EMAIL rgn5011@psu.edu PR:PHONE 7247662694 #STUDY ST:STUDY_TITLE Murine vitamin A deficiency results in a hypermetabolic state and alterations in ST:STUDY_TITLE bacterial community structure and metabolism.(Urine) ST:STUDY_SUMMARY Vitamin A deficiency (A-) is a significant public health problem. To better ST:STUDY_SUMMARY understand how vitamin A status influences gut microbiota and host metabolism, ST:STUDY_SUMMARY we systematically analyzed urine, cecum, serum, and liver samples from vitamin A ST:STUDY_SUMMARY sufficient (A+) and A- mice using 1H NMR-based metabolomics, quantitative ST:STUDY_SUMMARY (q)PCR, and 16S rRNA gene sequencing coupled with multivariate data analysis. ST:STUDY_SUMMARY The microbiota in the cecum of A- mice showed compositional as well as ST:STUDY_SUMMARY functional shifts compared to the microbiota from A+ mice. Targeted 1H NMR ST:STUDY_SUMMARY analyses revealed significant changes in microbial metabolite concentrations ST:STUDY_SUMMARY including higher butyrate and hippurate and decreased acetate and ST:STUDY_SUMMARY 4-hydroxyphenylacetate in A+ relative to A- mice. Bacterial butyrate-producing ST:STUDY_SUMMARY genes including butyryl-CoA:acetate CoA-transferase and butyrate kinase were ST:STUDY_SUMMARY significantly higher in bacteria from A+ versus bacteria from A- mice. A - mice ST:STUDY_SUMMARY had disturbances in multiple metabolic pathways including alterations in energy ST:STUDY_SUMMARY metabolism (hyperglycemia, glycogenesis, TCA cycle, and lipoprotein ST:STUDY_SUMMARY biosynthesis) and the A- host showed metabolites indicative of a hypermetabolic ST:STUDY_SUMMARY state (higher levels of amino acids and nucleic acids). A- mice had ST:STUDY_SUMMARY hyperglycemia, liver dysfunction, changes in bacterial metabolism, and altered ST:STUDY_SUMMARY gut microbial communities. Moreover, integrative analyses indicated a strong ST:STUDY_SUMMARY correlation between gut microbiota and host energy metabolism pathways in the ST:STUDY_SUMMARY liver. Vitamin A regulates the microbiota, bacterial metabolism and the effects ST:STUDY_SUMMARY of vitamin A on the microbiota results in alterations to host metabolism. ST:INSTITUTE Pennsylvania State University ST:LAST_NAME Nichols ST:FIRST_NAME Robert ST:ADDRESS 101 Life science building, University park, PA, 16803 ST:EMAIL rgn5011@psu.edu ST:PHONE 17247662694 #SUBJECT SU:SUBJECT_TYPE mouse SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS Vit A sufficient C1 Treatment:Control Genotype=Wild-Type SUBJECT_SAMPLE_FACTORS Vit A sufficient C2 Treatment:Control Genotype=Wild-Type SUBJECT_SAMPLE_FACTORS Vit A sufficient C3 Treatment:Control Genotype=Wild-Type SUBJECT_SAMPLE_FACTORS Vit A sufficient C4 Treatment:Control Genotype=Wild-Type SUBJECT_SAMPLE_FACTORS Vit A sufficient C5 Treatment:Control Genotype=Wild-Type SUBJECT_SAMPLE_FACTORS Vit A sufficient C6 Treatment:Control Genotype=Wild-Type SUBJECT_SAMPLE_FACTORS Vit A deficient T1 Treatment:Vit-A deficient food Genotype=Wild-Type SUBJECT_SAMPLE_FACTORS Vit A deficient T2 Treatment:Vit-A deficient food Genotype=Wild-Type SUBJECT_SAMPLE_FACTORS Vit A deficient T3 Treatment:Vit-A deficient food Genotype=Wild-Type SUBJECT_SAMPLE_FACTORS Vit A deficient T4 Treatment:Vit-A deficient food Genotype=Wild-Type SUBJECT_SAMPLE_FACTORS Vit A deficient T5 Treatment:Vit-A deficient food Genotype=Wild-Type SUBJECT_SAMPLE_FACTORS Vit A deficient T6 Treatment:Vit-A deficient food Genotype=Wild-Type #COLLECTION CO:COLLECTION_SUMMARY urine #TREATMENT TR:TREATMENT_SUMMARY Twelve male litters were weaned at 3 wks and continuously fed the vitamin A TR:TREATMENT_SUMMARY sufficient diet, vitamin A deficient diet, or the vitamin A deficient diet, TR:TREATMENT_SUMMARY supplemented with retenoic acid until the end of the experiment. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY The NMR sample prep for the urine, liver, cecal contents and serum are attached. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE - CH:INSTRUMENT_NAME - CH:COLUMN_NAME - #ANALYSIS AN:ANALYSIS_TYPE NMR #NMR NM:INSTRUMENT_NAME Bruker Avance III NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 1D-1H NM:SPECTROMETER_FREQUENCY 600 Mhz #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS total peak intensity per metabolite NMR_METABOLITE_DATA_START Samples C1 C2 C3 C4 C5 C6 T1 T2 T3 T4 T5 T6 Factors Treatment:Control Treatment:Control Treatment:Control Treatment:Control Treatment:Control Treatment:Control Treatment:Vit-A deficient food Treatment:Vit-A deficient food Treatment:Vit-A deficient food Treatment:Vit-A deficient food Treatment:Vit-A deficient food Treatment:Vit-A deficient food hippurate 250.6731656 202.6968613 196.6267142 216.9695819 179.4161928 239.9963858 198.7595936 203.3415659 167.9743074 142.8332548 172.679867 189.3842227 PAG 509.5759996 444.5828599 421.1258148 448.7129039 607.7959892 494.2317124 524.3036306 507.2351136 400.9022055 380.950891 447.2819154 447.7920189 IS 281.3477471 170.3647059 209.6309101 198.8115116 266.6106797 217.3031097 311.5927087 283.605262 287.2129253 172.141199 230.6551593 223.5808028 4-HPA 294.9234518 265.3563264 232.230322 242.3776327 447.1892373 270.7525755 209.6754708 294.7596491 186.6210147 236.8927987 245.4384213 201.2817186 TMAO 7.886985229 6.370776033 6.760731579 7.859306449 7.065138793 7.948923333 8.031412518 9.267394979 8.06568439 7.2022188 7.789678872 6.889986525 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name hippurate PAG IS 4-HPA TMAO METABOLITES_END #END