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Arteriovenous Blood Metabolomics: A Readout of Intra-Tissue Metabostasis.

Ivanisevic J, Elias D, Deguchi H, Averell PM, Kurczy M, Johnson CH, Tautenhahn R, Zhu Z, Watrous J, Jain M, Griffin J, Patti GJ, Siuzdak G - Sci Rep (2015)

Bottom Line: Global profiling of paired arterial and venous plasma from 20 healthy individuals, followed up by targeted analysis made it possible to measure subtle (<2 fold), yet highly statistically significant and physiologically important differences in water soluble human plasma metabolome.While we detected changes in lactic acid, alanine, glutamine, and glutamate as expected from skeletal muscle activity, a number of unanticipated metabolites were also determined to be significantly altered including Krebs cycle intermediates, amino acids that have not been previously implicated in transport, and a few oxidized fatty acids.This study provides the most comprehensive assessment of metabolic changes in the blood during circulation to date and suggests that such profiling approach may offer new insights into organ homeostasis and organ specific pathology.

View Article: PubMed Central - PubMed

Affiliation: Center for Metabolomics and Mass Spectrometry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, United States.

ABSTRACT
The human circulatory system consists of arterial blood that delivers nutrients to tissues, and venous blood that removes the metabolic by-products. Although it is well established that arterial blood generally has higher concentrations of glucose and oxygen relative to venous blood, a comprehensive biochemical characterization of arteriovenous differences has not yet been reported. Here we apply cutting-edge, mass spectrometry-based metabolomic technologies to provide a global characterization of metabolites that vary in concentration between the arterial and venous blood of human patients. Global profiling of paired arterial and venous plasma from 20 healthy individuals, followed up by targeted analysis made it possible to measure subtle (<2 fold), yet highly statistically significant and physiologically important differences in water soluble human plasma metabolome. While we detected changes in lactic acid, alanine, glutamine, and glutamate as expected from skeletal muscle activity, a number of unanticipated metabolites were also determined to be significantly altered including Krebs cycle intermediates, amino acids that have not been previously implicated in transport, and a few oxidized fatty acids. This study provides the most comprehensive assessment of metabolic changes in the blood during circulation to date and suggests that such profiling approach may offer new insights into organ homeostasis and organ specific pathology.

No MeSH data available.


Related in: MedlinePlus

Simultaneous measurement of metabolite levels in human arterial and venous blood.Two key gluconeogenic metabolites whose levels changed significantly after the blood passage through the forearm skeletal muscle. The changes revealed by untargeted metabolite profiling (p < 0.0001, Wilcoxon matched pairs rank test) were identified as glutamate and lactate, respectively, by MS/MS matching against standards in the METLIN database and validated by targeted analysis (paired plots). EIC—Extracted Ion Chromatogram. MS/MS—Tandem mass spectra or fragmentation pattern. Four stars—p value < 0.0001. Michael Kurczy and Julijana Ivanisevic have illustrated the blood circulation using Adobe Illustrator.
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f3: Simultaneous measurement of metabolite levels in human arterial and venous blood.Two key gluconeogenic metabolites whose levels changed significantly after the blood passage through the forearm skeletal muscle. The changes revealed by untargeted metabolite profiling (p < 0.0001, Wilcoxon matched pairs rank test) were identified as glutamate and lactate, respectively, by MS/MS matching against standards in the METLIN database and validated by targeted analysis (paired plots). EIC—Extracted Ion Chromatogram. MS/MS—Tandem mass spectra or fragmentation pattern. Four stars—p value < 0.0001. Michael Kurczy and Julijana Ivanisevic have illustrated the blood circulation using Adobe Illustrator.

Mentions: Untargeted metabolite profiling was performed initially to examine the differences between arterial and venous plasma across human forearm tissue, collected from healthy adult individuals (female:male 1:1) at rest, after an overnight fast. The population cohort description, including the gender, age, height, weight and body mass index, is given in the Supplemental Table S2. The approach using hydrophilic interaction chromatography coupled with negative ionization mode mass spectrometry based on 80% methanol extraction was applied to maximize the coverage of central carbon metabolites in human plasma (Fig. 1). The analytical strategy using HILIC in negative ionization mode enabled the detection of 8811 metabolite features in the water soluble and lipid plasma metabolome. Overall, untargeted profiling of arterial and venous plasma revealed high inter-individual variability with specific metabolic phenotypes characterizing each individual. These personalized metabotypes may interfere with profile alignment and comparisons, therefore paired analysis of the arterial and venous blood were essential when conducting global (untargeted) profiling of human blood. Many nutrients and metabolic by-products transported in plasma were analyzed across all subjects, including amino and non-amino organic acids, purines and pyrimidines, endogenous sugars, fatty acids, and dietary metabolites and their breakdown products (e.g. tagatose, paraxanthine). The global profiling followed by comparison of paired metabolite levels (measured simultaneously in the same individual) highlighted the significant changes (p < 0.01) in venous vs. arterial plasma during circulation across the human peripheral tissue, precisely the skeletal muscle (Fig. 2). Characteristic metabolites with significantly different levels in venous vs. arterial plasma were filtered out using the interactive XCMS Online platform and identified using MS/MS pattern matching against METLIN - the largest standard metabolite database (Fig. 1)2526. Among significantly changing metabolites, several metabolites involved in gluconeogenesis body cycle displayed highly consistent ratios between arterial and venous levels. Glutamate showed a significant decrease in venous vs. arterial plasma (p = 3.8E-06, median fold change = 2.9, Fig. 3) across all 20 subjects. Lactate demonstrated consistent difference characterized by significant increase in venous plasma (p = 4.2E-04, median fold change = 1.4, Fig. 3). In addition, two variants of oxidized unsaturated fatty acids, putatively identified as hydroxyperoxy octadecadienoic acids (HPODEs), were found highly increased in venous plasma ([M-H]− = 311.2224 and [M-H]− = 309.2067, with p = 9.7E-04 and 4.2E-04, respectively, and a median fold change > 30). The exact structure of these specific metabolites was not confirmed by further analysis, due to many potential variants and the absence of MS/MS matching hits in available databases (METLIN, HMDB, and LIPID MAPS)272829.


Arteriovenous Blood Metabolomics: A Readout of Intra-Tissue Metabostasis.

Ivanisevic J, Elias D, Deguchi H, Averell PM, Kurczy M, Johnson CH, Tautenhahn R, Zhu Z, Watrous J, Jain M, Griffin J, Patti GJ, Siuzdak G - Sci Rep (2015)

Simultaneous measurement of metabolite levels in human arterial and venous blood.Two key gluconeogenic metabolites whose levels changed significantly after the blood passage through the forearm skeletal muscle. The changes revealed by untargeted metabolite profiling (p < 0.0001, Wilcoxon matched pairs rank test) were identified as glutamate and lactate, respectively, by MS/MS matching against standards in the METLIN database and validated by targeted analysis (paired plots). EIC—Extracted Ion Chromatogram. MS/MS—Tandem mass spectra or fragmentation pattern. Four stars—p value < 0.0001. Michael Kurczy and Julijana Ivanisevic have illustrated the blood circulation using Adobe Illustrator.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4525490&req=5

f3: Simultaneous measurement of metabolite levels in human arterial and venous blood.Two key gluconeogenic metabolites whose levels changed significantly after the blood passage through the forearm skeletal muscle. The changes revealed by untargeted metabolite profiling (p < 0.0001, Wilcoxon matched pairs rank test) were identified as glutamate and lactate, respectively, by MS/MS matching against standards in the METLIN database and validated by targeted analysis (paired plots). EIC—Extracted Ion Chromatogram. MS/MS—Tandem mass spectra or fragmentation pattern. Four stars—p value < 0.0001. Michael Kurczy and Julijana Ivanisevic have illustrated the blood circulation using Adobe Illustrator.
Mentions: Untargeted metabolite profiling was performed initially to examine the differences between arterial and venous plasma across human forearm tissue, collected from healthy adult individuals (female:male 1:1) at rest, after an overnight fast. The population cohort description, including the gender, age, height, weight and body mass index, is given in the Supplemental Table S2. The approach using hydrophilic interaction chromatography coupled with negative ionization mode mass spectrometry based on 80% methanol extraction was applied to maximize the coverage of central carbon metabolites in human plasma (Fig. 1). The analytical strategy using HILIC in negative ionization mode enabled the detection of 8811 metabolite features in the water soluble and lipid plasma metabolome. Overall, untargeted profiling of arterial and venous plasma revealed high inter-individual variability with specific metabolic phenotypes characterizing each individual. These personalized metabotypes may interfere with profile alignment and comparisons, therefore paired analysis of the arterial and venous blood were essential when conducting global (untargeted) profiling of human blood. Many nutrients and metabolic by-products transported in plasma were analyzed across all subjects, including amino and non-amino organic acids, purines and pyrimidines, endogenous sugars, fatty acids, and dietary metabolites and their breakdown products (e.g. tagatose, paraxanthine). The global profiling followed by comparison of paired metabolite levels (measured simultaneously in the same individual) highlighted the significant changes (p < 0.01) in venous vs. arterial plasma during circulation across the human peripheral tissue, precisely the skeletal muscle (Fig. 2). Characteristic metabolites with significantly different levels in venous vs. arterial plasma were filtered out using the interactive XCMS Online platform and identified using MS/MS pattern matching against METLIN - the largest standard metabolite database (Fig. 1)2526. Among significantly changing metabolites, several metabolites involved in gluconeogenesis body cycle displayed highly consistent ratios between arterial and venous levels. Glutamate showed a significant decrease in venous vs. arterial plasma (p = 3.8E-06, median fold change = 2.9, Fig. 3) across all 20 subjects. Lactate demonstrated consistent difference characterized by significant increase in venous plasma (p = 4.2E-04, median fold change = 1.4, Fig. 3). In addition, two variants of oxidized unsaturated fatty acids, putatively identified as hydroxyperoxy octadecadienoic acids (HPODEs), were found highly increased in venous plasma ([M-H]− = 311.2224 and [M-H]− = 309.2067, with p = 9.7E-04 and 4.2E-04, respectively, and a median fold change > 30). The exact structure of these specific metabolites was not confirmed by further analysis, due to many potential variants and the absence of MS/MS matching hits in available databases (METLIN, HMDB, and LIPID MAPS)272829.

Bottom Line: Global profiling of paired arterial and venous plasma from 20 healthy individuals, followed up by targeted analysis made it possible to measure subtle (<2 fold), yet highly statistically significant and physiologically important differences in water soluble human plasma metabolome.While we detected changes in lactic acid, alanine, glutamine, and glutamate as expected from skeletal muscle activity, a number of unanticipated metabolites were also determined to be significantly altered including Krebs cycle intermediates, amino acids that have not been previously implicated in transport, and a few oxidized fatty acids.This study provides the most comprehensive assessment of metabolic changes in the blood during circulation to date and suggests that such profiling approach may offer new insights into organ homeostasis and organ specific pathology.

View Article: PubMed Central - PubMed

Affiliation: Center for Metabolomics and Mass Spectrometry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, United States.

ABSTRACT
The human circulatory system consists of arterial blood that delivers nutrients to tissues, and venous blood that removes the metabolic by-products. Although it is well established that arterial blood generally has higher concentrations of glucose and oxygen relative to venous blood, a comprehensive biochemical characterization of arteriovenous differences has not yet been reported. Here we apply cutting-edge, mass spectrometry-based metabolomic technologies to provide a global characterization of metabolites that vary in concentration between the arterial and venous blood of human patients. Global profiling of paired arterial and venous plasma from 20 healthy individuals, followed up by targeted analysis made it possible to measure subtle (<2 fold), yet highly statistically significant and physiologically important differences in water soluble human plasma metabolome. While we detected changes in lactic acid, alanine, glutamine, and glutamate as expected from skeletal muscle activity, a number of unanticipated metabolites were also determined to be significantly altered including Krebs cycle intermediates, amino acids that have not been previously implicated in transport, and a few oxidized fatty acids. This study provides the most comprehensive assessment of metabolic changes in the blood during circulation to date and suggests that such profiling approach may offer new insights into organ homeostasis and organ specific pathology.

No MeSH data available.


Related in: MedlinePlus