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Multiparametric Tumor Organoid Drug Screening Using Widefield Live-Cell Imaging for Bulk and Single-Organoid Analysis

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Successive High-Resolution (H₂O)-GCIB and C₆₀-SIMS Imaging Integrates Multi-Omics in Different Cell Types in Breast

Hua Tian¹, Louis J Sparvero^2,3, Tamil Selvan Anthonymuthu^2,4,3

¹Department of Chemistry, Pennsylvania State University, Chemistry Building, Shortlidge Rd, University Park, Pennsylvania 16802, United States.

Analytical Chemistry

|June 2, 2021

Summary

New mass spectrometry methods enable detailed lipid and metabolite analysis of single cells within the tumor microenvironment (TME). This approach reveals cellular communication and reprogramming crucial for understanding tumor progression and immune response.

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Area of Science:

Biotechnology
Mass Spectrometry Imaging
Cancer Research

Background:

Understanding the tumor microenvironment (TME) requires analyzing cell interactions and communication.
Current multi-omics profiling methods struggle with phenotypically diverse cells in native tissues.
New technologies are needed for simultaneous biomolecular profiling of individual cells within the TME.

Purpose of the Study:

To develop a novel methodology for comprehensive lipidomic and metabolomic profiling of individual cells within the TME.
To enable simultaneous analysis of biomolecular profiles and cell types on the same tissue section.
To investigate the role of lipid reprogramming and metabolic changes in tumor progression and immune suppression.

Main Methods:

Developed a dual mass spectrometry imaging approach using water gas cluster ion beam secondary ion mass spectrometry ((H2O)-GCIB-SIMS) for lipidomic and metabolomic profiling.
Utilized C60-SIMS with lanthanide antibodies for cell-type specific profiling on the same frozen-hydrated tissue sections.
Achieved high spatial resolution (1.6 μm for (H2O)-GCIB-SIMS, 1.1 μm for C60-SIMS) for single-cell analysis.

Main Results:

Successfully profiled >150 key ions, including lipids and metabolites, at the single-cell level within the TME.
Observed distinct variations in ion distribution and intensity across different cell types (e.g., tumor cells, immune cells).
Demonstrated the feasibility of integrating multi-omics profiling within a single tissue section at the single-cell resolution.

Conclusions:

The developed SIMS imaging technique allows for unprecedented multi-omics analysis of individual cells in their native tissue context.
This approach provides new insights into lipid reprogramming and metabolic responses driving cell-cell interactions in the TME.
The methodology has the potential to advance our understanding of normal tissue regulation and pathogenic processes.