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1.3 Gut Microbiome

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1.3 Gut Microbiome TECHNISCHE UNIVERSITÄT MÜNCHEN Lehrstuhl für Analytische Lebensmittelchemie Mass spectrometry based gut meta-metabolomics in obesity and type 2 Diabetes Alesia Walker Vollständiger Ausdruck der von der Fakultät für Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt der Technischen Universität München zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften genehmigter Dissertation. Vorsitzender: Univ.-Prof. Dr. E. Grill Prüfer der Dissertation: 1. apl.-Prof. Dr. Ph. Schmitt-Kopplin 2. Univ.-Prof. Dr. M. Rychlik 3. apl.-Prof. Dr. A. Hartmann, (Ludwig-Maximilians-Universität München) Die Dissertation wurde am 14.10.2013 bei der Technischen Universität München eingereicht und durch die Fakultät für Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt am 25.05.2014 angenommen. TABLE OF CONTENTS Table of contents Table of contents .................................................................................................................................................... I List of Figures ...................................................................................................................................................... IV List of Tables ......................................................................................................................................................... X Abbreviations ...................................................................................................................................................... XI Danksagung ....................................................................................................................................................... XII Summary ................................................................................................................................................................ a Zusammenfassung ................................................................................................................................................. b Chapter I ................................................................................................................................................................ 3 1 General Introduction ................................................................................................................................... 3 1.1 Metabolomics ................................................................................................................................... 3 1.2 Technologies in metabolomics ......................................................................................................... 5 1.2.1 Mass Spectrometry ............................................................................................................................ 5 1.2.1.1 Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) ................................. 6 1.2.1.2 Time of flight mass spectrometry (TOF-MS) ................................................................................... 7 1.3 Gut microbiome ................................................................................................................................ 9 1.3.1 Application of metabolomics to study the impact of gut microbiome ............................................ 11 1.3.2 Gut microbial metabolites ............................................................................................................... 12 1.3.2.1 The prediction of KEGG meta - metabolome ................................................................................. 14 1.3.3 Co - microbial metabolism in diseases ............................................................................................ 17 1.4 Main Objectives .............................................................................................................................. 20 Chapter II ............................................................................................................................................................ 25 2 Metabolomics in Type 2 Diabetes ............................................................................................................. 25 2.1 Introduction .................................................................................................................................... 25 2.2 Overview – Goals ........................................................................................................................... 27 2.2.1 Study design: Drug challenge in db/db mice ................................................................................... 27 2.3 Results and Discussion ................................................................................................................... 28 2.3.1 General: Body weight and blood glucose ........................................................................................ 28 2.3.2 The role of meta-metabolome in db/db mice during drug challenge ............................................... 29 2.3.2.1 Cholesterol and primary bile acid metabolism in db/db mice – metabolite patterns in the intestine 34 2.3.3 Differences of the fecal metabolome between db/db and wt mice .................................................. 38 2.3.3.1 KEGG metabolic pathway analysis of fecal meta-metabolome ..................................................... 40 2.3.4 Comparative analyses of metabolite classes between db/db and wt mice ....................................... 41 2.3.4.1 Fatty acids comparison in db/db mice vs. wt mice ......................................................................... 41 2.3.4.2 Oxylipins changes in db/db mice .................................................................................................... 44 2.3.4.3 N-acyltaurines in db/db mice .......................................................................................................... 45 2.3.4.4 Bile acids patterns in db/db mice .................................................................................................... 48 2.3.4.5 Arachidonic acid metabolism in db/db mice .................................................................................. 50 2.3.5 Mass difference analyses to reveal arachidonic acid co - metabolites applying NetCalc ................ 53 2.3.5.1 KEGG metabolic pathway enrichment analysis – sulfate conjugation ........................................... 56 2.3.6 Novel sulfur containing metabolites in db/db mice ......................................................................... 57 2.3.6.1 Sulfate conjugated metabolites: oxylipins sulfates ......................................................................... 57 2.3.6.2 Taurine conjugated metabolites: oxylipins taurines........................................................................ 59 2.3.6.3 Other N-acyl Fatty acids with amino acids ..................................................................................... 59 2.3.7 Plasma changes between db/db and wt mice based on KEGG metabolic pathway analyses .......... 60 2.3.8 Correlation studies and mass difference analyses between fecal and plasma samples of db/db mice ........................................................................................................................................................ 62 2.3.8.1 Correlation studies between feces and plasma samples .................................................................. 62 2.3.8.2 Mass difference analyses between fecal and plasma samples ........................................................ 64 2.3.9 Comparative analyses of fecal metabolome patterns between db/db and wt mice using UPLC-TOF- MS .................................................................................................................................................. 67 I 2.3.10 Metabolome analyses of COMBI treatment in db/db mice ............................................................. 68 2.3.11 Topographical Variation in wt mice ................................................................................................ 70 2.4 Summary and Conclusion ............................................................................................................... 73 Chapter III ........................................................................................................................................................... 77 3 Metabolomics in Obesity ............................................................................................................................ 77 3.1 Introduction .................................................................................................................................... 77 3.2 Overview – Goals ........................................................................................................................... 78 3.3 Study design of diet induced obesity (DIO) ................................................................................... 79 3.4 Results and Discussion ................................................................................................................... 79 3.4.1 General – Body weight changes ...................................................................................................... 79 3.4.2 Global analysis of cecal meta-metabolome due to DIO .................................................................. 80 3.4.3 Correlation studies: metabolites and body weight changes ............................................................
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