86 High throughput spatial mulitomic assay for assessing immune cell phenotype and function in the tumor microenvironment

• DOI: 10.1136/jitc-2024-sitc2024.0086
• Publication Date: 2024-11-05T14:58:59Z
• Authors: Debia Wakhloo; Julia Yu; Sonali A Deshpande; Anushka Dikshit

<h3>Background</h3> Tumormicroenvironment (TME) is a complex milieu of multiple cell types mainly including tumor cells, immune endothelial cells and stromal cells. Immune profile within the TME can determine success immunotherapies, indicate therapeutic efficacy as well any potential toxicity. Spatially interrogating these crucial in studying immune-immune immune-tumor interactions. Characterizing require assessing marker expression using protein detection activation markers RNA detection. To address this, we have developed high throughput RNAscope™ assay on BOND RX to spatially visualize same slide. <h3>Methods</h3> We utilized new Multiomic LS that detect up 6 targets The TSA-based amplification strategy offers signal boost for both targets. optimize detection, workflow completely protease-free there by preserving antigen integrity tissue morphology. be customized include target proteins interest. Here, demonstrate use our pre-conjugated antibody panel which includes CD8, CD4, FoxP3 PanCK infiltrating lymphocytes (TILs). also unconjugated primary antibodies CD68 CD163 macrophages tissues. <h3>Results</h3> Infiltrating Cytotoxic T were detected GZMB IFNG co-expression. Regulatory FOXP3 was used delineate from stroma. Similarly, associated CD163, , IL10 IL-1B expression. able identify distinct M1 M2 macrophage populations samples. <h3>Conclusions</h3> RNAscope successfully interrogate different subpopulation with quality while maintaining higher plex capability provides more simultaneously, turn enabling phenotype profiling sensitivity specificity technology known for.