Translational cancer research has seen an increasing interest in metabolomic profiling to decipher tumor phenotypes. However, the impact of post-surgical freezing delays on mass spectrometric metabolomic measurements of the cancer tissue remains elusive.
To evaluate the impact of post-surgical freezing delays on cancer tissue metabolomics and to investigate changes per metabolite and per metabolic pathway.
We performed untargeted metabolomics on three cortically located and bulk-resected glioblastoma tissues that were sequentially frozen as duplicates at up to six different time delays (0–180 min, 34 samples).
Statistical modelling revealed that 10% of the metabolome (59 of 597 metabolites) changed significantly after a 3 h delay. While carbohydrates and energy metabolites decreased, peptides and lipids increased. After a 2 h delay, these metabolites had changed by as much as 50–100%. We present the first list of metabolites in glioblastoma tissues that are sensitive to post-surgical freezing delays and offer the opportunity to define individualized fold change thresholds for future comparative metabolomic studies.
More researchers should take these pre-analytical factors into consideration when analyzing metabolomic data. We present a strategy for how to work with metabolites that are sensitive to freezing delays.
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Data and software availability
The metabolomic data has been processed and analyzed using the recently released MetaboDiff R package (Mock et al. 2018). MetaboDiff is platform-independent, available at http://github.com/andreasmock/MetaboDiff/ and has been released under the MIT license. The data of this paper, including a step-by-step user guide explaining the preprocessing and statistical analyses performed, can be found at http://github.com/andreasmock/quenching/.
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We thank Evelin Eiswirth for her help in visualizing the study design. Furthermore, we thank Farzaneh Kashfi, Ilka Hearn, Melanie Greibich, and Mandy Barthel for their excellent technical assistance.
Conflict of interest
The authors declare no conflict of interest.
The study was approved by the ethics committee of the Medical Faculty, University of Heidelberg (reference number: 005/2003) and written informed consent was obtained from all patients in accordance with the Declaration of Helsinki and its later amendments.
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Mock, A., Rapp, C., Warta, R. et al. Impact of post-surgical freezing delay on brain tumor metabolomics. Metabolomics 15, 78 (2019). https://doi.org/10.1007/s11306-019-1541-2