Multimodal Mass Spectrometry Imaging of an Osteosarcoma Multicellular Tumour Spheroid Model to Investigate
Sophie M Pearce1, Neil A Cross1, David P Smith1
1Centre for Mass Spectrometry Imaging, Biomolecular Sciences Research Centre, Sheffield Hallam University, Howard Street, Sheffield S1 1WB, UK.
Abstract:
A multimodal mass spectrometry imaging (MSI) approach was used to investigate the chemotherapy drug-induced response of a Multicellular Tumour Spheroid (MCTS) 3D cell culture model of osteosarcoma (OS). The work addresses the critical demand for enhanced translatable early drug discovery approaches by demonstrating a robust spatially resolved molecular distribution analysis in tumour models following chemotherapeutic intervention. Advanced high-resolution techniques were employed, including desorption electrospray ionisation (DESI) mass spectrometry imaging (MSI), to assess the interplay between metabolic and cellular pathways in response to chemotherapeutic intervention. Endogenous metabolite distributions of the human OS tumour models were complemented with subcellularly resolved protein localisation by the detection of metal-tagged antibodies using Imaging Mass Cytometry (IMC). The first application of matrix-assisted laser desorption ionization-immunohistochemistry (MALDI-IHC) of 3D cell culture models is reported here. Protein localisation and expression following an acute dosage of the chemotherapy drug doxorubicin demonstrated novel indications for mechanisms of region-specific tumour survival and cell-cycle-specific drug-induced responses. Previously unknown doxorubicin-induced metabolite upregulation was revealed by DESI-MSI of MCTSs, which may be used to inform mechanisms of chemotherapeutic resistance. The demonstration of specific tumour survival mechanisms that are characteristic of those reported for in vivo tumours has underscored the increasing value of this approach as a tool to investigate drug resistance.
Insights
This study used multimodal mass spectrometry imaging (MSI) to analyze osteosarcoma multicellular tumor spheroids treated with doxorubicin. The findings reveal new insights into drug resistance mechanisms and metabolite changes in 3D cancer models.
Area of Science:
- Oncology
- Biochemistry
- Analytical Chemistry
Background:
- Multicellular tumor spheroids (MCTS) offer a 3D model for studying cancer drug responses.
- Osteosarcoma (OS) drug discovery requires improved, translatable preclinical models.
- Understanding chemotherapy resistance mechanisms is crucial for effective treatment.
Purpose of the Study:
- To investigate chemotherapy drug-induced responses in osteosarcoma MCTS using a multimodal mass spectrometry imaging (MSI) approach.
- To analyze the spatial distribution of metabolites and proteins in response to doxorubicin treatment.
- To explore novel mechanisms of tumor survival and drug resistance in 3D cancer models.
Main Methods:
- Utilized desorption electrospray ionization mass spectrometry imaging (DESI-MSI) for endogenous metabolite distribution.
- Employed Imaging Mass Cytometry (IMC) for subcellular protein localization using metal-tagged antibodies.
- Applied matrix-assisted laser desorption ionization-immunohistochemistry (MALDI-IHC) to 3D cell culture models for the first time.
Main Results:
- Identified region-specific tumor survival mechanisms and cell-cycle-specific drug responses to doxorubicin.
- Discovered previously unknown doxorubicin-induced metabolite upregulation via DESI-MSI.
- Demonstrated protein localization and expression changes following chemotherapeutic intervention.
Conclusions:
- The multimodal MSI approach provides robust, spatially resolved molecular analysis of 3D tumor models.
- Findings offer insights into osteosarcoma chemotherapeutic resistance mechanisms.
- This technique enhances the value of 3D models for investigating drug resistance in early drug discovery.
