NMR and MS-based Stable Isotope-Resolved Metabolomics and applications in cancer metabolism

Abstract

There is considerable interest in defining metabolic reprogramming in human diseases, which is recognized as a hallmark of human cancer. Although radiotracers have a long history in specific metabolic studies, stable isotope-enriched precursors coupled with modern high resolution mass spectrometry and NMR spectroscopy have enabled systematic mapping of metabolic networks and fluxes in cells, tissues and living organisms including humans. These analytical platforms are high in information content, are complementary and cross-validating in terms of compound identification, quantification, and isotope labeling pattern analysis of a large number of metabolites simultaneously. Furthermore, new developments in chemoselective derivatization and in vivo spectroscopy enable tracking of labile/low abundance metabolites and metabolic kinetics in real-time. Here we review developments in Stable Isotope Resolved Metabolomics (SIRM) and recent applications in cancer metabolism using a wide variety of stable isotope tracers that probe both broad and specific aspects of cancer metabolism required for proliferation and survival.

Resolving Metabolic Heterogeneity in Experimental Models of the Tumor Microenvironment from a Stable Isotope Resolved Metabolomics Perspective

“…Stable isotopes especially 13 C, 15 N, and 2 H (D) are fully biologically compatible and isotopically enriched substrates can be administered with no or minimal biological effects. Stable isotope tracing of metabolic pathways has been demonstrated in a wide variety of biological systems [32,[91][92][93][94][95][96][97] for the reconstruction of metabolic networks [98][99][100] and also, in some cases, mapping of metabolic flux [77,90,91,[101][102][103][104][105][106].…”

“…We have begun to develop the above described approaches to investigate metabolic interactions between human lung cancer cells and Mφ in spheroid co-cultures. We have also introduced mSIRM [97,107,121,170] in our studies to maximize information retrieval from limited samples and to minimize batch artifacts. The following describes some preliminary findings that illustrate the value of these approaches.…”

Metabolomic Approaches to Study Chemical Exposure-Related Metabolism Alterations in Mammalian Cell Cultures

“…As life is built upon carbon chemistry, carbon-13 is a stable isotope tracer frequently used in NMR and MS metabolomics studies [ 69 , 70 , 71 ]. In NMR, individual 13C atoms can be tracked through isotopomers, i.e., isomers that differ only by the position of 13C, and followed over chemical transformations of metabolic pathways.…”

“…In fluxomics experiments, culture cells are put in contact with 13C-labelled substrates, such as glucose, and 13C labeled product metabolites are quantified either by NMR or MS. This permits a very precise quantification of fluxes of matter along metabolic pathways [ 70 , 72 ]. Using this approach, direct impacts of a perturbation, be it a chemical added to the cell culture or a genetic mutation, can be identified.…”

Metabolomics Reveals the Alteration of Metabolic Pathway by Alpha-Melanocyte-Stimulating Hormone in B16F10 Melanoma Cells

“…Metabolomics is a very powerful tool that provide a more comprehensive understanding of the effects of biological potential at a specified time in particular environmental conditions [ 12 ]. With the development in the last few decades of analytical instruments, such as mass spectrometry and nuclear magnetic resonance spectroscopy, metabolomic analysis of cells has been widely used to perform a complete assessment of functional cellular responses to pathogen infections, toxicity and environmental factors [ 13 ]. In addition, studies have reported that metabolites directly initiate cell signaling stages and regulate various biological processes such as post-translational modifications and epigenetic mechanisms [ 14 ].…”