Application of stable isotope labels for metabolomics in studies in fatty liver disease

Patrycja Puchalska, Peter Crawford

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The progression of nonalcoholic fatty liver disease (NAFLD) increases the risks of cirrhosis and cardiovascular disease. Marked alteration of both cytosolic and mitochondrial metabolism, and in combination with insulin resistance, increases hepatic glucose production. Utilization of stable isotope tracers to study liver metabolism offers deep insight into rearrangements of metabolic pathways and substrate-product relationships under the conditions leading to fatty liver and induced by diseases, drugs, toxins, or genetic manipulations. Isotope tracing untargeted metabolomics (ITUM) recently emerged as a powerful platform in which the label can be tracked in an untargeted fashion, revealing the penetration of substrates into metabolic pathways, even at low abundance. Here, we describe a protocol that can be utilized to study the changes in utilization of any labeled substrate toward a wide range of metabolites either in isolated liver cells or whole liver tissue under conditions mimicking various stages of fatty liver disease. Furthermore, a routine protocol for extraction, separation, and mass spectrometric detection of isotopically labeled metabolites in an untargeted or targeted fashion. An informatic approach to analyze stable isotope untargeted metabolomic datasets is also described.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages259-272
Number of pages14
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1996
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Fingerprint

Metabolomics
Fatty Liver
Isotopes
Liver Diseases
Liver
Metabolic Networks and Pathways
Informatics
Insulin Resistance
Fibrosis
Cardiovascular Diseases
Glucose
Pharmaceutical Preparations

Keywords

  • Fatty liver disease
  • Isotope tracking untargeted metabolomics
  • Mass spectrometry
  • Stable isotopes
  • Untargeted metabolomics

Cite this

Puchalska, P., & Crawford, P. (2019). Application of stable isotope labels for metabolomics in studies in fatty liver disease. In Methods in Molecular Biology (pp. 259-272). (Methods in Molecular Biology; Vol. 1996). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9488-5_20

Application of stable isotope labels for metabolomics in studies in fatty liver disease. / Puchalska, Patrycja; Crawford, Peter.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 259-272 (Methods in Molecular Biology; Vol. 1996).

Research output: Chapter in Book/Report/Conference proceedingChapter

Puchalska, P & Crawford, P 2019, Application of stable isotope labels for metabolomics in studies in fatty liver disease. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1996, Humana Press Inc., pp. 259-272. https://doi.org/10.1007/978-1-4939-9488-5_20
Puchalska P, Crawford P. Application of stable isotope labels for metabolomics in studies in fatty liver disease. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 259-272. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9488-5_20
Puchalska, Patrycja ; Crawford, Peter. / Application of stable isotope labels for metabolomics in studies in fatty liver disease. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 259-272 (Methods in Molecular Biology).
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