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Stable isotope resolved metabolomics of lung cancer in a SCID mouse model

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Abstract

We have determined the time course of [U-13C]-glucose utilization and transformations in SCID mice via bolus injection of the tracer in the tail vein. Incorporation of 13C into metabolites extracted from mouse blood plasma and several tissues (lung, heart, brain, liver, kidney, and skeletal muscle) were profiled by NMR and GC–MS, which helped ascertain optimal sampling times for different target tissues. We found that the time for overall optimal 13C incorporation into tissue was 15–20 min but with substantial differences in 13C labeling patterns of various organs that reflected their specific metabolism. Using this stable isotope resolved metabolomics (SIRM) approach, we have compared the 13C metabolite profile of the lungs in the same mouse with or without an orthotopic lung tumor xenograft established from human PC14PE6 lung adenocarcinoma cells. The 13C metabolite profile shows considerable differences in [U-13C]-glucose transformations between the two lung tissues, demonstrating the feasibility of applying SIRM to investigate metabolic networks of human cancer xenograft in the mouse model.

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Abbreviations

BHT:

Butylated hydroxytoluene

DSS:

2,2′Dimethylsilapentane-5-sulfonate

MTBSTFA:

N-methyl-N-[tert-butyl-dimethylsilyl]trifluoroacetamide

NSCLC:

Non small cell lung cancer

SIRM:

Stable isotope resolved metabolomics

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Acknowledgments

This work was supported in part by National Science Foundation EPSCoR grant # EPS-0447479, NIH Grant Number P20RR018733 from the National Center for Research Resources, 1R01CA118434-01A2 (to TWMF), R01CA-086412 and RO1 CA150947 (to JY) from the National Cancer Institute, the Kentucky Challenge for Excellence, Susan G. Komen Foundation BCTR0503648 and NCI R21CA133688 (to ANL), and the Brown Foundation. We thank J. Tan, Ruifeng Su and Mr. Richard Hansen for technical assistance. We also thank the two reviewers for many useful comments and additional references to improve the manuscript.

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Authors and Affiliations

  1. Department of Chemistry, University of Louisville, 2210 S. Brook St, Rm 348 John W. Shumaker Research Building, Louisville, KY, 40292, USA

    Teresa W.-M. Fan, Andrew N. Lane & Richard M. Higashi

  2. Department of Medicine, James Graham Brown Cancer Center, Clinical Translational Research Building, 505 S. Hancock St., Louisville, KY, 40202, USA

    Teresa W.-M. Fan, Andrew N. Lane & Jun Yan

  3. Center for Regulatory Environmental Metabolomics, University of Louisville, 2210 S. Brook St., Louisville, KY, 40292, USA

    Teresa W.-M. Fan, Andrew N. Lane & Richard M. Higashi

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  1. Teresa W.-M. Fan
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  2. Andrew N. Lane
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  4. Jun Yan
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Correspondence to Teresa W.-M. Fan.

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Fan, T.WM., Lane, A.N., Higashi, R.M. et al. Stable isotope resolved metabolomics of lung cancer in a SCID mouse model. Metabolomics 7, 257–269 (2011). https://doi.org/10.1007/s11306-010-0249-0

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