SciLifeLab Science Summit - SB

Spatial biology techniques have revolutionized our understanding of the tumor microenvironment (TME) and its intricate cellular interactions. Multiplex immunofluorescence (mIF) methods enable precise profiling of immune cells and other key players within the TME, revealing their spatial distribution and interactions (PMID: 38012408). In situ hybridization (ISH) technologies complement protein profiling by mapping cytokine- and chemokine-expressing cells, crucial for deciphering signaling networks and immune activation.  

In this context, a new multiomics method that integrates RNAscope™ and sequential immunofluorescence (seqIF™) protocols is presented, allowing co-detection of RNAs and proteins within the TME by using COMET™, an automated tissue imaging and staining platform. The integrated protocol allows up to 12 RNA and 24 protein biomarker detection cycles, generating a 12-plex RNA and 24-plex protein panel without human intervention.  

We validated the assay sensitivity and specificity and developed a panel of 12 probes to target essential RNA biomarkers of tumor-infiltrating lymphocytes and their activation states in human FFPE tumor tissues. Simultaneously, we employed a 24-antibody panel to detect protein biomarkers to enable single-cell identification within the TME. Applying this approach to human FFPE tumor tissues, we demonstrated that co-detection of RNA and protein biomarkers in the same section enhances our understanding of key cellular components involved in tumor progression and immune response. This integrated approach promises deeper insights into cancer biology.  

Our findings underscore the promise of deeper insights into tissue biology with spatial multiomics technologies. By fully automating these technologies on platforms like COMET™, we improve the throughput of spatial multiomics assay, reduce user interventions, and increase the robustness of assays. This progress has important implications for creating predictive markers, refining cancer diagnoses, and tailoring personalized therapies.