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Molecular Profiling of Glioblastoma Patient-Derived Single Cells Using Combined MALDI-MSI and MALDI-IHC
Kasper K Krestensen1, Tim F E Hendriks1, Andrej Grgic1
1The Maastricht MultiModal Molecular Imaging (M4I) institute, Division of Imaging Mass Spectrometry (IMS), Maastricht University, 6229 ER Maastricht, The Netherlands.
This study validates matrix-assisted laser desorption/ionization–immunohistochemistry (MALDI-IHC) for single-cell analysis, combining it with mass spectrometry imaging (MSI) to reveal cell-specific lipid profiles and protein markers. The workflow accurately classifies glioblastoma cells, offering new insights into tumor heterogeneity.
Area of Science:
- Utilizes advanced mass spectrometry imaging (MSI) techniques for high-resolution molecular profiling.
- Integrates matrix-assisted laser desorption/ionization–immunohistochemistry (MALDI-IHC) for multiplexed protein visualization.
- Focuses on single-cell analysis within complex biological samples like glioblastoma.
Background:
- Mass spectrometry-based imaging, particularly MALDI-MSI, has advanced significantly in spatial resolution, enabling single-cell molecular profiling.
- MALDI-immunohistochemistry (MALDI-IHC) allows multiplexed protein visualization in tissues but lacks validation for single-cell cultures.
- Understanding cell-to-cell interactions is crucial for deciphering intratumor heterogeneity.
Purpose of the Study:
- To introduce and validate a workflow combining MALDI-MSI and MALDI-IHC for single, isolated cells.
- To investigate cell type-specific lipid profiles and protein markers in patient-derived glioblastoma cells.
- To assess the compatibility of initial MSI measurements with subsequent MALDI-IHC analysis.
Main Methods:
- Developed a workflow for sequential MALDI-MSI and MALDI-IHC on single, isolated glioblastoma and neuronal cells.
- Performed high-resolution MSI for untargeted lipid profiling.
- Applied MALDI-IHC for targeted protein marker visualization.
- Created an automated recognition model based on MALDI-MSI data for cell classification.
Main Results:
- The multimodal imaging successfully revealed distinct lipid profiles differentiating glioblastoma cells from neuronal cells.
- Initial MSI measurement and sample preparation did not negatively impact subsequent MALDI-IHC ion intensities.
- An automated model accurately classified cells based on lipid profiles (triglycerides, phosphatidylcholines, sphingomyelins) consistent with MALDI-IHC markers.
Conclusions:
- MALDI-IHC provides valuable molecular information in single-cell measurements, even after initial MSI analysis, without efficacy loss.
- This validated workflow enhances the understanding of molecular complexities at the single-cell level.
- The approach offers unique insights into cell-to-cell interactions driving intratumor heterogeneity, paving the way for future research.
