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Metabolomic and lipidomic characterization of Oxalobacter formigenes strains HC1 and OxWR by UHPLC-HRMS

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Abstract

Diseases of oxalate, such as nephrolithiasis and primary hyperoxaluria, affect a significant portion of the US population and have limited treatment options. Oxalobacter formigenes, an obligate oxalotrophic bacterium in the mammalian intestine, has generated great interest as a potential probiotic or therapeutic treatment for oxalate-related conditions due to its ability to degrade both exogenous (dietary) and endogenous (metabolic) oxalate, lowering the risk of hyperoxaluria/hyperoxalemia. Although all oxalotrophs degrade dietary oxalate, Oxalobacter formigenes is the only species shown to initiate intestinal oxalate secretion to draw upon endogenous, circulating oxalate for consumption. Evidence suggests that Oxalobacter regulates oxalate transport proteins in the intestinal epithelium using an unidentified secreted bioactive compound, but the mechanism of this function remains elusive. It is essential to gain an understanding of the biochemical relationship between Oxalobacter and the host intestinal epithelium for this microbe to progress as a potential remedy for oxalate diseases. This investigation includes the first profiling of the metabolome and lipidome of Oxalobacter formigenes, specifically the human strain HC1 and rat strain OxWR, the only two strains shown thus far to initiate net intestinal oxalate secretion across native gut epithelia. This study was performed using untargeted and targeted metabolomics and lipidomics methodologies utilizing ultra-high-performance liquid chromatography-mass spectrometry. We report our findings that the metabolic profiles of these strains, although largely conserved, show significant differences in their expression of many compounds. Several strain-specific features were also detected. Discussed are trends in the whole metabolic profile as well as in individual features, both identified and unidentified.

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Abbreviations

BMP:

Bis(monoacylglycero)phosphate

CER:

Ceramide

CL:

Cardiolipin

CoQ10:

Coenzyme Q10

DG:

Diacylglycerol

DMPE:

Dimethylphosphatidylethanolamine

FA:

Formic acid

LPC:

Lysophosphatidylcholine

LPE:

Lysophosphatidylethanolamine

LSM:

Lysosphingomyelin

OxPE:

Oxidized phosphatidylethanolamine

PAzePC:

1-Palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine

PC:

Phosphatidylcholine

PE:

Phosphatidylethanolamine

PG:

Phosphatidylglycerol

PI:

Phosphatidylinositol

PS:

Phosphatidylserine

SM:

Sphingomyelin

SO:

Sphingosine

TG:

Triacylglycerol

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Acknowledgments

The authors would like to acknowledge Dr. Cory A. Leonard, Department of Pathology, Immunology and Laboratory Medicine, University of Florida, for her assistance with sample generation for this experiment, including media preparation and cell culture, harvest, and lysis. Also to be acknowledged are Dr. Jeremy P. Koelmel, Department of Pathology, Immunology and Laboratory Medicine, University of Florida, and Vanessa Y. Rubio, Department of Chemistry, University of Florida, for their contributions with figure generation for the lipidomics analysis and graphical abstract, respectively.

Funding

This work was funded by the National Institutes of Health grant 2R01DK088892-05A1.

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  1. Department of Pathology, Immunology and Laboratory Medicine, University of Florida, Gainesville, FL, 32610, USA

    Casey A. Chamberlain, Marguerite Hatch & Timothy J. Garrett

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  1. Casey A. Chamberlain
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  2. Marguerite Hatch
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Correspondence to Timothy J. Garrett.

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Published in the topical collection Young Investigators in (Bio-)Analytical Chemistry with guest editors Erin Baker, Kerstin Leopold, Francesco Ricci, and Wei Wang.

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Chamberlain, C.A., Hatch, M. & Garrett, T.J. Metabolomic and lipidomic characterization of Oxalobacter formigenes strains HC1 and OxWR by UHPLC-HRMS. Anal Bioanal Chem 411, 4807–4818 (2019). https://doi.org/10.1007/s00216-019-01639-y

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