Technology information

Description
Simultaneous Multiplexed Imaging of mRNA and Proteins with Subcellular Resolution in Breast Cancer Tissue Samples by Mass Cytometry. To build comprehensive models of cellular states and interactions in normal and diseased tissue, genetic and proteomic information must be extracted with single-cell and spatial resolution. Here, we extended imaging mass cytometry to enable multiplexed detection of mRNA and proteins in tissues. Three mRNA target species were detected by RNAscope-based metal in situ hybridization with simultaneous antibody detection of 16 proteins. Analysis of 70 breast cancer samples showed that HER2 and CK19 mRNA and protein levels are moderately correlated on the single-cell level, but that only HER2, and not CK19, has strong mRNA-to-protein correlation on the cell population level. The chemoattractant CXCL10 was expressed in stromal cell clusters, and the frequency of CXCL10-expressing cells correlated with T cell presence. Our flexible and expandable method will allow an increase in the information content retrieved from patient samples for biomedical purposes, enable detailed studies of tumor biology, and serve as a tool to bridge comprehensive genomic and proteomic tissue analysis.

Comment
Mass spectrometry‐based method is one of the highly multiplex techniques to capture the protein spatial intensity. IMC is another method dependent on metal‐tagged antibodies and enables the imaging of over 100 antibodies. Such method offers unprecedented opportunities to explore regional immunity composition and topological function units of TME. However, this Method dependent on the performance of antibodies and are relatively costly. It is still challenging to increase the current throughput to proteome‐wide. Bias may also exist when designing the panel of markers rather than discovering functional proteins from the proteomics data. [PMID: 35040595]

Keywords
Cell Line, Erbb-2, Genetics, Receptor, Analysis, Diagnostic Imaging, Tumor Microenvironment, Single-Cell Analysis, Cytology, In Situ Hybridization, Systems Biology, Physiology, Female, Proteomics, Tumor, Gene Expression Regulation, HeLa Cells, Messenger, Breast Neoplasms

Targets
Protein

Spatial resolution
Subcellular