Redox phospholipidomics discovers pro-ferroptotic death signals in A375 melanoma cells in vitro and in vivo

Yulia Y Tyurina; Alexandr A Kapralov; Vladimir A Tyurin; Galina Shurin; Andrew A Amoscato; Dhivyaa Rajasundaram; Hua Tian; Yuri L Bunimovich; Yulia Nefedova; William G Herrick; Ralph E Parchment; James H Doroshow; Hulya Bayir; Apurva K Srivastava; Valerian E Kagan

doi:10.1016/j.redox.2023.102650

Redox phospholipidomics discovers pro-ferroptotic death signals in A375 melanoma cells in vitro and in vivo

Redox Biol. 2023 May:61:102650. doi: 10.1016/j.redox.2023.102650. Epub 2023 Feb 28.

Authors

Affiliations

¹ Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA; Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA, USA. Electronic address: yyt1@pitt.edu.
² Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA; Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA, USA.
³ Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA; Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA.
⁴ Department of Critical Care Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
⁵ Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA; Department of Dermatology, University of Pittsburgh, Pittsburgh, PA, USA.
⁶ The Wistar Institute, Philadelphia, PA, USA.
⁷ Clinical Pharmacodynamic Biomarkers Program, Applied/Developmental Research Directorate, Frederick National Laboratory for Cancer Research, Frederick, MD, USA.
⁸ Division of Cancer Treatment and Diagnosis, National Cancer Institute, Bethesda, MD, USA.
⁹ Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA; Department of Critical Care Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
¹⁰ Clinical Pharmacodynamic Biomarkers Program, Applied/Developmental Research Directorate, Frederick National Laboratory for Cancer Research, Frederick, MD, USA. Electronic address: srivastavaa4@mail.nih.gov.
¹¹ Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA; Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA, USA. Electronic address: kagan@pitt.edu.

Abstract

Growing cancer cells effectively evade most programs of regulated cell death, particularly apoptosis. This necessitates a search for alternative therapeutic modalities to cause cancer cell's demise, among them - ferroptosis. One of the obstacles to using pro-ferroptotic agents to treat cancer is the lack of adequate biomarkers of ferroptosis. Ferroptosis is accompanied by peroxidation of polyunsaturated species of phosphatidylethanolamine (PE) to hydroperoxy- (-OOH) derivatives, which act as death signals. We demonstrate that RSL3-induced death of A375 melanoma cells in vitro was fully preventable by ferrostatin-1, suggesting their high susceptibility to ferroptosis. Treatment of A375 cells with RSL3 caused a significant accumulation of PE-(18:0/20:4-OOH) and PE-(18:0/22:4-OOH), the biomarkers of ferroptosis, as well as oxidatively truncated products - PE-(18:0/hydroxy-8-oxo-oct-6-enoic acid (HOOA) and PC-(18:0/HOOA). A significant suppressive effect of RSL3 on melanoma growth was observed in vivo (utilizing a xenograft model of inoculation of GFP-labeled A375 cells into immune-deficient athymic nude mice). Redox phospholipidomics revealed elevated levels of 18:0/20:4-OOH in RSL3-treated group vs controls. In addition, PE-(18:0/20:4-OOH) species were identified as major contributors to the separation of control and RSL3-treated groups, with the highest variable importance in projection predictive score. Pearson correlation analysis revealed an association between tumor weight and contents of PE-(18:0/20:4-OOH) (r = -0.505), PE-18:0/HOOA (r = -0.547) and PE 16:0-HOOA (r = -0.503). Thus, LC-MS/MS based redox lipidomics is a sensitive and precise approach for the detection and characterization of phospholipid biomarkers of ferroptosis induced in cancer cells by radio- and chemotherapy.

Keywords: A375 melanoma cells; Biomarkers; Ferroptosis; Hydroperoxy-phosphatidylethanolamine; Redox lipidomics.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Cell Death
Chromatography, Liquid
Humans
Lipid Peroxidation
Melanoma*
Mice
Mice, Nude
Oxidation-Reduction
Tandem Mass Spectrometry*

Abstract

Publication types

MeSH terms

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