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Differential lipid analysis of oxaliplatin-sensitive and resistant HCT116 cells reveals different levels of drug-induced lipid droplet formation

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Abstract

Lipid droplets (LDs) are intracellular storage vesicles composed of a neutral lipid core surrounded by a glycerophospholipid membrane. LD accumulation is associated with different stages of cancer progression and stress responses resulting from chemotherapy. In previous work, a novel dual nano-electrospray ionization source and data-dependent acquisition method for measuring the relative abundances of lipid species between two extracts were described and validated. Here, this same source and method were used to determine if oxaliplatin-sensitive and resistant cells undergo similar lipid profile changes, with the goal of identifying potential signatures that could predict the effectiveness of an oxaliplatin-containing treatment. Oxaliplatin is commonly used in the treatment of colorectal cancer. When compared to a no-drug control, oxaliplatin dosing caused significant increases in triglyceride (TG) and cholesterol ester (CE) species. These increases were more pronounced in the oxaliplatin-sensitive cells than in oxaliplatin-resistant cells. The increased neutral lipid abundance correlated with LD formation, as confirmed by confocal micrographs of Nile Red–stained cells. Untargeted proteomic analyses also support LD formation after oxaliplatin treatment, with an increased abundance of LD-associated proteins in both the sensitive and resistant cells.

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Acknowledgements

We thank the UNC Microscopy Services Laboratory (MSL) and its director, Dr. Pablo Ariel, for confocal microscope training and access. We also thank Dr. Cameron Worthington and Ms. Alexis Zimmer for their helpful discussions.

Funding

This work was supported by the National Institute of General Medical Sciences through Grant Award Number R35 GM128697. Some of this research was conducted at the UNC Proteomics Core Facility and Microscopy Services Laboratory (MSL). These facilities are supported in part by an NCI Center Core Support Grant (P30 CA016086), awarded to the UNC Lineberger Comprehensive Cancer Center.

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  1. Department of Chemistry, University of North Carolina at Chapel Hill, Kenan and Caudill Laboratories, Chapel Hill, NC, 27599-3290, USA

    Tyler S. Larson, Thomas J. DiProspero, Gary L. Glish & Matthew R. Lockett

  2. Lineberger Comprehensive Cancer Center, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599-7295, USA

    Matthew R. Lockett

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Correspondence to Gary L. Glish or Matthew R. Lockett.

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The authors declare the following competing financial interest(s): Bruker Daltonics maintains an active license to some of the previously patented UNC DIMS IP used in this work.

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Larson, T.S., DiProspero, T.J., Glish, G.L. et al. Differential lipid analysis of oxaliplatin-sensitive and resistant HCT116 cells reveals different levels of drug-induced lipid droplet formation. Anal Bioanal Chem 416, 151–162 (2024). https://doi.org/10.1007/s00216-023-05010-0

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