Nuclear Magnetic Resonance Spectroscopy to Identify Metabolite Biomarkers of Nonresponsiveness to Targeted Therapy in Glioblastoma Tumor Stem Cells

• DOI: 10.1021/acs.jproteome.8b00801
• Publication Date: 2019-04-09T20:37:51Z
• Authors: Ingvild Comfort H...; Henriette Engen Berg; Daniel Sachse; Erlend Skaga; Frøydis Sved Skot...; Elsa Lundanes; Cecilie J. Sandberg; Einar O. Vik-Mo; Frode Rise; Steven Ray Wilson
• Keywords: 0301 basic medicine; Principal Component Analysis; Magnetic Resonance Spectroscopy; Brain Neoplasms; Cell Survival; Survivin; Citric Acid Cycle; Primary Cell Culture; Imidazoles; Antineoplastic Agents; Biomarkers, Pharmacological; Citric Acid; 3. Good health; 03 medical and health sciences; Metabolome; Neoplastic Stem Cells; Humans; Lactic Acid; Molecular Targeted Therapy; Glioblastoma; Naphthoquinones

Glioblastoma is the most common and malignant brain tumor, and current therapies confer only modest survival benefits. A major obstacle is our ability to monitor treatment effect on tumors. Current imaging modalities are ambiguous, and repeated biopsies are not encouraged. To scout for markers of treatment response, we used NMR spectroscopy to study the effects of a survivin inhibitor on the metabolome of primary glioblastoma cancer stem cells. Applying high resolution NMR spectroscopy (1H resonance frequency: 800.03 MHz) to just 3 million cells per sample, we achieved sensitive and high resolving determinations of, e.g., amino acids, nucleosides, and constituents of the citric acid cycle. For control samples that were cultured, prepared, and measured at varying dates, peak area relative standard deviations were 15-20%. Analyses of unfractionated lysates were performed for straightforward compound identification with COLMAR and HMDB databases. Principal component analysis revealed that citrate levels were clearly upregulated in nonresponsive cells, while lactate levels substantially decreased following treatment for both responsive and nonresponsive cells. Hence, lactate and citrate may be potential markers of successful drug uptake and poor response to survivin inhibitors, respectively. Our metabolomics approach provided alternative biomarker candidates compared to spectrometry-based proteomics, underlining benefits of complementary methodologies. These initial findings make a foundation for exploring in vivo MR spectroscopy (MRS) of brain tumors, as citrate and lactate are MRS-visible. In sum, NMR metabolomics is a tool for addressing glioblastoma.