Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Application of the HILIC-MS Novel Protocol to Study the Metabolic Heterogeneity of Glioblastoma Cells

Version 1 : Received: 9 April 2024 / Approved: 10 April 2024 / Online: 11 April 2024 (13:45:06 CEST)

How to cite: Šofranko, J.; Gondáš, E.; Murín, R. Application of the HILIC-MS Novel Protocol to Study the Metabolic Heterogeneity of Glioblastoma Cells. Preprints 2024, 2024040751. https://doi.org/10.20944/preprints202404.0751.v1 Šofranko, J.; Gondáš, E.; Murín, R. Application of the HILIC-MS Novel Protocol to Study the Metabolic Heterogeneity of Glioblastoma Cells. Preprints 2024, 2024040751. https://doi.org/10.20944/preprints202404.0751.v1

Abstract

Glioblastoma is a highly malignant brain tumor consisting of heterogeneous cellular population. The transformed metabolism of glioblastoma cells supports their growth and division on the background of their milieu. One might hypothesize that the transformed metabolism of a primary glioblastoma could be well adapted to limitations in the variety and number of substrates imported into the brain parenchyma and present it their microenvironment. Additionally, the phenotypic heterogeneity of cancer cells could promote the variations among their metabolic capabilities regarding the utilization of available substrates and release of metabolic intermediates. With the aim to identify the putative metabolic footprint of different types of glioblastoma cells, we exploited the possibility for separation of polar and ionic molecules present in culture media or cell lysates by hydrophilic interaction liquid chromatography (HILIC). The mass spectrometry (MS) was then used to identify and quantify the eluted compounds. The introduced method allows the detection and quantification of more than 150 polar and ionic metabolites in single run, those may be present either in culture media or cell lysates and provide data for polaromic studies within metabolomics. The method was applied to analyze the culture media and cell lysates derived from two types of glioblastoma cells, T98G and U118. The analysis revealed that even the both types of glioblastoma cells share several common metabolic aspects, they also exhibit differences in their metabolic capability. This finding agrees with the hypothesis about metabolic heterogeneity of glioblastoma cells. Furthermore, the combination of both analytical methods, HILIC-MS, provides a valuable tool for metabolomic studies based on the simultaneous identification and quantification of a wide range of polar and ionic metabolites - polaromics.

Keywords

glioblastoma; HILIC; LC-MS; metabolomics; amino acid; metabolic heterogeneity

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

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