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Mapping chemotherapeutic drug distribution in cancer cell spheroids using 2D-TOF-SIMS and LESA-TIMS-MS
Yarixa L Cintron-Diaz1, Arlet M Acanda de la Rocha, Anthony Castellanos
1Department of Chemistry and Biochemistry, Florida International University, 11200 SW 8th St., AHC4-233, Miami, FL 33199, USA. fernandf@fiu.edu.
Abstract:
Three-dimensional (3D) cancer cell cultures grown in the form of spheroids are effective models for the study of in vivo-like processes simulating cancer tumor pharmacological dynamics and morphology. In this study, we show the advantages of Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS) combined with in situ Liquid Extraction Surface Analysis coupled to trapped Ion Mobility Spectrometry Mass Spectrometry (LESA-TIMS-TOF MS) for high spatial resolution mapping and quantitation of ABT-737, a chemotherapeutic drug, at the level of single human colon carcinoma cell spheroids (HCT 116 MCS). 2D-TOF-SIMS studies of consecutive sections (∼16 μm thick slices) showed that ABT-737 is homogenously distributed in the outer layers of the HCT 116 MCS. Complementary in situ LESA-TIMS-TOF MS/MS measurements confirmed the presence of the ABT-737 drug in the MCS slides by the observation of the molecular ion [M + H]+m/z and mobility, and the charateristic fragmentation pattern. LESA-TIMS-TOF MS allowed a quantitative assessment of the ABT-737 drug of the control MCS slice spiked with ABT-737 standard over the 0.4-4.1 ng range and MCS treated starting at 10 μM for 24 h. These experiments showcase an effective protocol for unambigous characterization and 3D mapping of chemotherapeutic drug distribution at the single MCS level.
Insights
This study demonstrates Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS) and LESA-TIMS-TOF MS for mapping chemotherapeutic drug distribution in 3D cancer spheroids. The methods provide high spatial resolution for drug quantitation in single cell cultures.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Cancer Research
Background:
- Three-dimensional (3D) cancer cell spheroids mimic in vivo tumor characteristics, aiding pharmacological studies.
- Accurate drug distribution mapping in spheroids is crucial for understanding treatment efficacy.
- Advanced mass spectrometry techniques offer potential for high-resolution analysis of drug localization.
Purpose of the Study:
- To showcase the utility of TOF-SIMS combined with LESA-TIMS-TOF MS for high spatial resolution mapping and quantitation of chemotherapeutic drugs.
- To analyze the distribution of the drug ABT-737 within single human colon carcinoma cell spheroids (HCT 116 MCS).
- To establish an effective protocol for unambiguous characterization and 3D mapping of drug distribution at the single spheroid level.
Main Methods:
- Utilized Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS) for 2D mapping of drug distribution in spheroid sections.
- Employed in situ Liquid Extraction Surface Analysis coupled to trapped Ion Mobility Spectrometry Mass Spectrometry (LESA-TIMS-TOF MS) for drug confirmation and quantitation.
- Analyzed consecutive ∼16 μm thick slices of HCT 116 MCS treated with ABT-737.
Main Results:
- 2D-TOF-SIMS revealed homogeneous distribution of ABT-737 in the outer layers of HCT 116 MCS.
- LESA-TIMS-TOF MS/MS confirmed ABT-737 presence via molecular ion and fragmentation patterns.
- LESA-TIMS-TOF MS enabled quantitative assessment of ABT-737 in control and treated MCS over a defined concentration range.
Conclusions:
- The combined TOF-SIMS and LESA-TIMS-TOF MS approach provides high spatial resolution for drug mapping in 3D cancer spheroids.
- This methodology allows for unambiguous characterization and quantitation of chemotherapeutic drugs at the single spheroid level.
- The developed protocol is effective for studying drug pharmacodynamics and distribution in advanced cancer models.
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