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.tifRx=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.

空间交互映射在癌细胞、免疫细胞和基质细胞的相互作用中为患者分层及推进免疫治疗提供了新的机遇。尽管单细胞研究揭示了前列腺癌中的显著分子异质性，但目前尚不清楚免疫细胞的异质性如何影响局部肿瘤中肿瘤细胞与基质细胞之间的空间协调。本研究中，我们采用循环免疫荧光成像技术对整个组织切片进行成像，揭示了低级和高级前列腺癌及其邻近正常组织中癌细胞与基质细胞之间新的空间关联。我们的研究结果提供了一个包含699,461个单细胞的空间图谱，这些细胞在不同临床分级中显示出表观遗传学和分子差异。我们报道了一种独特的肥大细胞群体，该群体不同表达CD44、CD90和颗粒酶B（GZMB），并证明GZMB+肥大细胞在前列腺肿瘤中与M2巨噬细胞存在空间关联。最终，我们揭示了主要由雄激素受体阳性（AR+）基质细胞驱动的常见邻域，并确定了在AR+前列腺基质中活跃的转录网络。每个组织具有10轮成像数据；一轮淬灭成像数据（R3Q）。每轮成像由5个tif图像组成，来自5个通道（DAPI、M1、M2、M3和M4）。文件名结构：Rx_M1.M2.M3.M4_TissueID_SceneID_ChannelID_ORG.tif，其中Rx为轮次顺序（R1-R10）；M1为标记1；M2为标记2；M3为标记3；M4为标记4；TissueID为组织编号；SceneID为所有图像的单个场景（s001）；ChannelID为通道0-5；R3Q为R3成像后的淬灭轮。