Metabolic fingerprinting of chemotherapy-resistant prostate cancer stem cells. An untargeted metabolomic approach by liquid chromatography-mass spectrometry

Alicia Bort; Belén G Sánchez; Carlos León; Leonor Nozal; José M Mora-Rodríguez; Florentina Castro; Antonio L Crego; Inés Díaz-Laviada

doi:10.3389/fcell.2022.1005675

Metabolic fingerprinting of chemotherapy-resistant prostate cancer stem cells. An untargeted metabolomic approach by liquid chromatography-mass spectrometry

Front Cell Dev Biol. 2022 Oct 17:10:1005675. doi: 10.3389/fcell.2022.1005675. eCollection 2022.

Authors

Alicia Bort¹, Belén G Sánchez², Carlos León³, Leonor Nozal⁴, José M Mora-Rodríguez², Florentina Castro⁴, Antonio L Crego⁵, Inés Díaz-Laviada²

Affiliations

¹ Yale University School of Medicine, Vascular Biology and Therapeutics Program, New Haven, CT, United states.
² Alcala University, School of Medicine, Department of Systems Biology and Research Institute in Chemistry "Andrés M. Del Río" (IQAR), Madrid, Spain.
³ Carlos III University, Department of Bioengineering and Aerospatial Engineering, Madrid, Spain.
⁴ Alcala University and General Foundation of Alcalá University, Center of Applied Chemistry and Biotechnology, Madrid, Spain.
⁵ Alcala University, Department of Analytical Chemistry, Physical Chemistry and Chemical Engineering, Madrid, Spain.

Abstract

Chemoresistance is one of the most important challenges in cancer therapy. The presence of cancer stem cells within the tumor may contribute to chemotherapy resistance since these cells express high levels of extrusion pumps and xenobiotic metabolizing enzymes that inactivate the therapeutic drug. Despite the recent advances in cancer cell metabolism adaptations, little is known about the metabolic adaptations of the cancer stem cells resistant to chemotherapy. In this study, we have undertaken an untargeted metabolomic analysis by liquid chromatography-high-resolution spectrometry combined with cytotoxicity assay, western blot, quantitative real-time polymerase chain reaction (qPCR), and fatty acid oxidation in a prostate cancer cell line resistant to the antiandrogen 2-hydroxiflutamide with features of cancer stem cells, compared to its parental androgen-sensitive cell line. Metabolic fingerprinting revealed 106 out of the 850 metabolites in ESI+ and 67 out of 446 in ESI- with significant differences between the sensitive and the resistant cell lines. Pathway analysis performed with the unequivocally identified metabolites, revealed changes in pathways involved in energy metabolism as well as posttranscriptional regulation. Validation by enzyme expression analysis indicated that the chemotherapy-resistant prostate cancer stem cells were metabolically dormant with decreased fatty acid oxidation, methionine metabolism and ADP-ribosylation. Our results shed light on the pathways underlying the entry of cancer cells into dormancy that might contribute to the mechanisms of drug resistance.

Keywords: cancer chemoresistance; cancer stem cells; fatty acid oxidation; liquid chromatography-mass spectrometry; prostate cancer; untargeted metabolomic.