Figure 5.

Construction of the CRC immune module (CCIM)

(A) The cell-cell interaction (CCI) network for all cell subpopulations in CRC. The color of edges indicates the type of ligand cell, and the size of edges represents the CCI numbers between cells.

(B) The number of interactions between monocytes/macrophages and CD4⁺ T cells inferred from single-cell RNA sequencing (scRNA-seq).

(C) The number of interactions between monocytes/macrophages and CD8⁺ T cells.

(D) mIHC staining for FOLR2 (red), DKK3 (white), FOXP3 (yellow), PDCD1 (orange), CD4 (green), CD8 (light blue), and DAPI (blue) reveals the CCIM image pattern. Scale bar, 50 μm. The dotted line indicates the margin between tumor tissue and para-tumor tissue.

(E) The proportion of cells from CCIM and other cell types around FOLR2⁺ macrophages. The statistics were determined with the area of 800 pixels around FOLR2⁺ macrophages. Biological replicates, n = 5.

(F) Voronoi plot for the CCIM topological pattern. The green dot represents the center of each part.

(G) Depiction of the CCIM module. A CCIM module contains at least one FOLR2⁺ macrophage, one tolerant CD8⁺ T cell, one Treg cell, and one exhausted CD4⁺ T cell.

(H) The density plot for the CCIM in CRC.

(I) Heatmap of cell-type-specific receptor-ligand interactions inferred by SingleCellSignalR. Shown are inferred interactions between FOLR2⁺ macrophages and Treg cells, exhausted CD4⁺ T cells, exhausted CD4⁺ T cells, and tolerant CD8⁺ T cells. Circle color indicates the inferred interaction score, and circle size indicates the mean expression of receptor and ligand genes for each pair.

(J–L) The association of exhausted T cells (p < 0.01) (J), tolerant T cells (p <0.01) (K), and Treg cells (p<0.01) (L) with FOLR2⁺ macrophages. The p value was calculated with Pearson’s coefficient, and p < 0.05 was considered to indicate significance.

(M) The CCIM score stratifies CRC patients with poor prognosis (patients, n = 1,405).