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North Carolina Central UniversityLuke Chen, PhDNCCU-01-16VWDecember 27, 2016Author: Doug Lee, PhD

Metabolomic impact of Nrf2 activation on esophageal epithelium

Study OverviewStudy DesignStudy Objective

The goal of this study was to characterize the metabolic impact of loss of Nrf2 and Keap1 on mouse esophageal epithelium tissue and (in a separate adult wild type cohort) to identify changes in esophageal epithelial

metabolism.

This 30-sample study examined global metabolic profiles in mouse esophageal tissue from Nrf2 and Keap1

knockouts.

Global metabolic profiles were determined from the experimental groups outlined in the table below.

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Group Name

Group Description

N

WT

Wild-type

N=6

Nrf2 KO

Nrf2 knockout

N=6

Keap1 KO

Keap1 knockout

N=3

Principal Component Analysis

The samples separate

Glutathione, precursors and GSH biosynthetic pathway intermediates were enhanced in the Keap1KO and decreased in the NRF2 KO. These changes are consistent with Nrf2s role in regulating anti-oxidant defenses.

Redox Metabolism - Glutathione Metabolism

Bilirubin was increased in the Keap1KO and heme was increased in the Nrf2 KO, changes that are consistent with heme oxygenase as a substrate of Nrf2 (PMID: 21868703).Redox Metabolism - Heme Metabolism

Several cofactors and anti-oxidants were increased in the Keap1KO suggesting that Nfr2 enhances anti-oxidant defenses and metabolic homeostasis.

Redox Metabolism - Antioxidants and Cofactors

Several intermediates of the citric acid cycle were increased in the Keap1KO suggesting that Nrf2 enhances energy metabolism and/or mitochondrial function.Carbohydrate Metabolism - Energy

Intermediates related to glycogen were enhanced in the Keap1 KO suggesting Nrf2 enhances glycogen metabolismCarbohydrate Metabolism - Glycogen

Arabitol/xylitol was increased in Keap1KO and decreased in the Nrf2 KO; and several pentose phosphate pathway (PPP) intermediates and pentitol metabolites were increased in the Keap1 KO suggesting that Nrf2 channels metabolism through the PPP pathway.Carbohydrate Metabolism- PPP

Nucleotide and aminosugar intermediates, which are channeled from the glycolysis, were increased in Keap 1 KO but decreased in the NRF2KO, suggesting that glycosylation reactions are enhanced by Nrf2.Carbohydrate Metabolism - Nucleotide Sugars

Many phospholipids were increased in the Keap1 KO suggesting Nrf2 enhances lipid biosynthesis.Lipid Metabolism

Several acyl-carnitines and 3-hydroxybutyrate were increased in the Nrf2KO suggesting that Nrf2 downregulates beta-oxidation.

Lipid Metabolism - Beta Oxidation

Lipid Metabolism - SignalingEicosanoids and endocannabinoids including arachidonylethanolamide, were increased in Keap1 KO suggesting that Nrf2 enhances lipid signaling.

CMP and UMP were increased in the Keap1 KO and decreased in Nrf2 KO; many purine and pyrimidine metabolites were increased in Keap1KO suggesting that Nrf2 enhances nucleotide biosynthesis or nucleic acid turnover.Nucleic Acid Metabolism

Subpathway Analysis: WT vs NRF2KO (p<0.01)Subpathway

p value

Gamma-glutamyl Amino Acid

5.91E-05

Glutathione Metabolism

1.09E-04

Fatty Acid, Dihydroxy

2.00E-04

Nucleotide Sugar

2.29E-04

Disaccharides and Oligosaccharides

2.91E-04

Vitamin B6 Metabolism

8.31E-04

Pyrimidine Metabolism, Orotate containing

0.00199839

Mevalonate Metabolism

2.57E-03

Medium Chain Fatty Acid

0.00290864

Glycine, Serine and Threonine Metabolism

0.00557812

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Subpathway Analysis: WT vs KEAP1KO (p<0.01)Subpathway

p value

Nucleotide Sugar

3.46E-07

Pyrimidine Metabolism, Uracil containing

1.72E-06

Methionine, Cysteine, SAM and Taurine Metabolism

2.14E-06

Phosphatidylethanolamine (PE)

3.49E-06

Sphingolipid Metabolism

1.07E-05

Purine Metabolism, (Hypo)Xanthine/Inosine containing

1.31E-05

Aminosugar Metabolism

2.30E-05

Glutamate Metabolism

2.76E-05

Glycine, Serine and Threonine Metabolism

2.83E-05

Phosphatidylinositol (PI)

3.38E-05

Ceramides

3.73E-05

Phosphatidylglycerol (PG)

4.8473E-05

Phosphatidylserine (PS)

6.47E-05

Pentose Metabolism

7.77E-05

Sterol

8.92E-05

Folate Metabolism

0.000124461

Advanced Glycation End-product

0.000153135

Purine Metabolism, Adenine containing

1.67E-04

Histidine Metabolism

1.85E-04

Phosphatidylcholine (PC)

1.92E-04

Glutathione Metabolism

2.35E-04

Plasmalogen

2.60E-04

Hemoglobin and Porphyrin Metabolism

3.29E-04

Tryptophan Metabolism

3.68E-04

Pyrimidine Metabolism, Thymine containing

3.68E-04

Riboflavin Metabolism

4.34E-04

Ascorbate and Aldarate Metabolism

4.79E-04

Alanine and Aspartate Metabolism

5.50E-04

Pantothenate and CoA Metabolism

6.91E-04

Gamma-glutamyl Amino Acid

7.98E-04

Phenylalanine Metabolism

9.65E-04

Phospholipid Metabolism

9.81E-04

Fatty Acid Metabolism (Acyl Choline)

0.001180258

Fructose, Mannose and Galactose Metabolism

0.001218238

Pentose Phosphate Pathway

0.001670689

Purine Metabolism, Guanine containing

0.001677319

Glycogen Metabolism

0.001840044

Eicosanoid

0.002672507

Endocannabinoid

0.002786562

Bacterial/Fungal

0.003362365

TCA Cycle

0.003588187

Dipeptide Derivative

0.00377853

Pyrimidine Metabolism, Cytidine containing0.004465765Tyrosine Metabolism0.005150577Acetylated Peptides0.007691728Purine and Pyrimidine Metabolism0.00806993Medium Chain Fatty Acid0.009477024

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