Q-TOF LC/MS-based Untargeted Metabolomics Approach to Evaluate the Effect of Folate-Conjugated Cyclodextrins on Triple-Negative Breast Cancer Cells

Background: Breast cancer is a heterogenic disease that comprises of various morphologies with intrinsic subtypes and is principally responsible for casualties among female cancer patients. Triple-negative breast cancer (TNBC) is the most aggressive subtype with a high probability of relapsing, hence successful treatment can be quite challenging. The pathogenesis of TNBC remains ambiguous and the identification of dependable biomarkers for its early diagnosis is crucial to design a strategy for therapeutic armamentarium. Objective: To clarify the folate-dependent mechanism of action causing cell death and to unravel the potential biomarkers of TNBC to defeat this consequential public health burden. Methods: The MTT assay and the morphological examination via microscopy were carried out to examine the viability of the cells upon the administration of the blank folate-conjugated cyclodextrin nanoparticles. An untargeted metabolomic approach using Q-TOF LC/MS was performed. Multivariate analysis of the metabolomic profile was applied to the MDA-MB-231 cell line with the aim of comparing the untreated cells with the folate-conjugated cyclodextrin nanoparticles applied cells to detect possible biomarkers. Results: The spectrophotometric and microscopic analyses revealed that MDA-MB-231 cells underwent early apoptosis following the incubation with the folate-conjugated nanoparticles for 24 h of administration. Moreover, metabolomics profiling pointed out that hexose metabolism was significantly altered. Data mining procedures showed that glycolysis, mannose, fructose, and galactose were the most affected pathways in TNBC upon blank folate-conjugated cyclodextrin nanoparticle administration and this effect was determined to be cell-specific. A perturbed hexose pathway may be the explanation of selective cell death and decelerated cell growth seen in TNBC cells. Conclusion: Our study offers a new understanding of the underlying mechanisms of TNBC since we hereby provide evidence that hexose is one of the main driving forces for the metabolic mechanism over TNBC cells. This alternative mechanistic approach may markedly increase the effect of chemotherapy on TNBC.