Fig. 1: Spatial transcriptome atlas of human CRCs by Stereo-seq reveals ICB responsiveness.

a Schematic of single cell and spatial transcriptomic analysis in CRC patients with or without neoadjuvant anti-PD1 treatment. b The top differential express gene (DEG) expressions in each spatial cluster are shown in the matrix plot. c The box and whisker plot shows the proportions of spatial clusters tumor_MKI67 and tumor-stroma boundary in indicated patient groups. The smooth muscle is not included to remove sampling bias. Data are represented as mean±IQR and analyzed by unpaired 2-tailed Student-t test with Bonferroni correction. Ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ***, p < 0.0001. N number: pMMR=6; dMMR=4; dSD=2; dPR=1; dCR=3. d Representative images of FOV and H&E staining in treatment naïve pMMR (patient #59), and anti-PD1 treated dMMR with stable disease (SD) (patient #95). Low mRNA-enriched region (fiber and cavity) is not displayed. Scale bars were set at 1200 μm for the global spatial plot (left) and 300μm for the FOVs (middle and right). e The box and whisker plot shows the average distance from epi/tumor cluster to the clusters of tumor-stroma boundary, immune aggregates, stroma and smooth muscle. Data are represented as mean ± IQR and analyzed by unpaired 2-tailed Student t test with Bonferroni correction. Ns not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ***, p < 0.0001. N number of the spots: tumor-stroma boundary=17,639; immune aggregates=16,304; stroma=28,012; smooth muscle=24,172. f Pearson correlation between spatial clusters tumor_MKI67 and tumor-stroma boundary. The dot plot color represents indicated patient group. The line and the band present the linear regression model and confidence interval (95%) respectively. N number of the patients: pMMR=6; dMMR=4; dSD=2; dPR=1; dCR=3.