Metabolomics: LIPIDOMIC AND DESI IMAGING STUDY OF MOUSE LIVER DOSED WITH A TYROSINE KINASE INHIBITING DRUG

Importance of the Topic

Gefitinib is a tyrosine kinase inhibitor used in cancer therapy with known effects on lipid metabolism and potential liver toxicity. Detailed lipidomic profiling and spatial imaging of liver tissues can uncover drug induced molecular changes and support safer drug development.

Objectives and Study Overview

This study investigated lipid alterations in mouse liver following intravenous administration of Gefitinib at multiple time points. A combined approach of untargeted lipidomics, targeted LC-MS/MS assays and DESI imaging was used to capture both global lipidome changes and spatial distribution of drug and lipids.

Methodology and Instrumentation

Approaches:

• Untargeted lipidomics: Reversed-phase LC-IM-MS(TOF) on a C18 column with ion mobility separation. Data processed by Progenesis QI and MetaboAnalyst via PCA and PLS-DA models.
• Targeted lipidomics: HILIC LC-MS/MS (LipidQuan) using a library of over 2000 lipids. Quantitative assays developed with Quanpedia standards to monitor >500 biologically relevant lipids.
• DESI imaging: Cryosectioned liver slices (10 µm) analyzed under ambient conditions to map spatial distributions of Gefitinib and selected lipids. Data analysis performed with HDI and multivariate statistics.

Used Instrumentation

• UPLC systems with ACQUITY Premier CSH C18 and BEH Amide columns
• Xevo TQ-XS triple quadrupole mass spectrometer
• SYNAPT XS ion mobility QToF mass spectrometer
• DESI XS imaging source
• Software: Progenesis QI, LipoStar, MetaboAnalyst, HDI

Main Results and Discussion

• PCA and PLS-DA analyses demonstrated clear separation between control and dosed groups and revealed a time-dependent trajectory of lipidome changes across 0.5 to 24 hours.
• Key lipids such as LPE(18:0), PG(16:0/18:1) and PC(16:0/18:1) were identified as significant markers of Gefitinib exposure.
• Dysregulation of phospholipid and bioactive lipid pathways followed distinct temporal patterns post-dose.
• DESI imaging confirmed spatial accumulation of Gefitinib and correlated distribution shifts of key lipids within liver tissue.

Benefits and Practical Applications

• Broad lipid coverage facilitates biomarker discovery for drug induced liver effects.
• Rapid targeted assays reduce development time and enhance quantitation confidence.
• Ambient DESI imaging provides label-free spatial molecular information to complement bulk lipidomics.

Future Trends and Potential Applications

• Integration of collision cross section data into untargeted workflows to refine lipid identifications.
• Expansion of targeted lipid libraries for comprehensive bioactive lipid monitoring across platforms.
• Advances in ambient imaging MS for higher spatial resolution and multiplexed quantitation.
• Adoption of combined lipidomics and imaging strategies in clinical and regulatory settings for drug safety evaluation.

Conclusion

The combined application of untargeted lipidomics, targeted LC-MS/MS and DESI imaging reveals time-dependent alterations in liver lipid metabolism induced by Gefitinib and maps their spatial distributions. This integrated workflow offers a powerful platform for investigating drug effects on tissue lipidomes and supports biomarker identification and safety assessment.

References

1. McKillop D Hutchison M Partridge EA Bushby N Cooper CM Clarkson Jones JA Herron W Swaisland HC Xenobiotica 2005 34 914 934
2. Molloy BJ King A Mullin LG Gethings LA Riley R Plumb RS Wilson ID Xenobiotica 2021 51 434 446
3. Want E Masson P Michopoulos F Nat Protoc 2013 8 17 32
4. Chong J Soufan O Li C Caraus I Li S Bourque G Wishart DS Xia J Nucleic Acids Res 2018 46 W486 W494