Multiplex imaging of localized prostate tumors reveals changes in the spatial organization of AR-positive cells in the microenvironment

Mapping spatial interactions of cancer, immune and stromal cells present novel opportunities for patient stratification and for advancing immunotherapy. While single-cell studies revealed significant molecular heterogeneity in prostate tumors, there is currently no understanding of how immune cell heterogeneity impacts spatial coordination between tumor and stromal cells in localized tumors. Here, we used cyclic immunofluorescent imaging on whole-tissue sections to uncover novel spatial associations between cancer and stromal cells in low- and high-grade prostate tumors and tumor-adjacent normal tissues. Our results provide a spatial map of 699,461 single-cells that show epigenetic and molecular differences in distinct clinical grades. We report unique populations of mast cells that differentially express CD44, CD90 and Granzyme B (GZMB) and demonstrate GZMB+ mast cells are spatially associated with M2 macrophages in prostate tumors. Finally, we uncover recurrent neighborhoods that are primarily driven by androgen receptor positive (AR+) stromal cells and identify transcriptional networks active in AR+ prostate stroma.

Each tissue has 10 Rounds of imaging data; one quenching round of imaging data (R3Q). For each round of imaging consists of 5 tif images from 5 channels (DAPI, M1, M2, M3 and M4).

File Name structure: Rx_M1.M2.M3.M4_TissueID_SceneID_ChannelID_ORG.tif

Rx=Round order (R1-R10); M1=Marker 1; M2=Marker 2; M3=Marker 3; M4=Marker 4; TissueID=Tissue Number; SceneID=Single scene for all images (s001); ChannelID=Channels 0-5; R3Q=Quenching round after R3 imaging.