Figure 1 Myeloid lineage microenvironment between LCC and RCC
(A) t-SNE plot showing the transcriptome landscape of 370,115 cells from datasets of 62 CRC patients. Proportions of the global cells in left-sided and right-sided CRC were shown. (B), 39,167 myeloid cells from solid tumor tissues were extracted from the dataset. Porpotions of the myeloid lineages in left-sided and right-sided CRC were shown. (C) The fraction of the 4 myeloid subpopulations that originated from left-sided (blue) and right-sided (red) CRC samples were presented. (D) A volcano plot of DEGs (differential expressed genes) that were up-regulated (avg_logFC > 0.25, p-value < 0.05) or downregulated (avg_logFC < -0.25, p-value < 0.05) between left-sided and right-sided CRC. (E) Heatmap of all DEGs showed that tumorigenesis-asscoaited genes that were up-regulted in right-sided CRC, while immune-related genes that were up-regulated in left-sided CRC.
Figure S1. Heterogeneity of cell poprotions and biological functions between left-sided and right-sided CRC
Cell poprotions between left-sided and right-sided CRC from normal (A) and patient (tumor) samples. Visualization of the DEGs (left) and (right) DEPs (differentially expressed pathways) between left-sided and right-sided CRC from myeloid lineage of granulocytes (C), DC (D), Monocytes (E), and Macrophages (F).
Figure 2 neutrophils exhibit distinct functionalities between left-sided and right-sided CRC
(A) The UMAP plot showed the distribution of granulocytes. Granulocytes were reclustered into 4 subclusters. The fraction of cells htat originated from left-sided and right-sided CRC samples for 4 subgroups identified in this profile. (B) Expression profile of 5 granulocyte-migration-associated genes in macrophages. (C) Bubble plot showing the communication probability for selected ligand-receptor pairs. Dot size indicated P-value, colored by communication probability. Granulocytes-specific ligand-receptor pairs were highlighted. (D) Chord diagram showing preferential interations of granulocytes with various myeloid subclusters mediated by CCL23-CCR1 ligand-receptor pair. (E) Expression profile of signatures of granulocytes from left-sided (blue) and right-sided (red) CRC. The Kaplan-Meier overall survival curves of TCGA COAD patients grouped by the right-sided CRC (F) and left-sided CRC (G) signatures. (H) Expression of a antitumor signature across granulocytes from left-sided and right-sided CRC. Two-sided Wilcoxon test. antitumor signatures contain TNF, ICAM1, IFNB1, CCL3, TNFSF10, IL21. (I) Differential pathway enriched in granulocytes from left-sided and right-sided CRC.
Figure S2 Genes involved in granulocyte migration
UMAP visualization showing the expression of selected genes in macrophages of left-sided and right-sided CRC.
Figure 3 Heterogeneity and dynamics of DC subsets between left-sided and right-sided CRC
(A) The t-SNE plot showing the distribution of DCs. DCs were clustered into 7 subclusters. The fraction of cells that originated from left-sided and right-sided CRC samples for 7 subgroups identified in this profile. (B) Number of up-regulated genes in each DC subset between left-sided and right-sided CRC. (C) Expression of CCL3 and CCR1 in left-sided and right-sided CRC. (D) Violin plot showing the ISG (IFN-stimulated genes) score across DC subsets in CRC. ISG signatures, MX2, ISG15, IRF7, BST2, IFITM2, and IFI27. (E) Heatmap of DEGs and DEPs in DC subsets (padj < 0.01, and avg_logFC > 0.25). The expression of each gene was centered to the average expression across DC subsets with a scale from -2 to 2. (F). Correlations of all DC subsets. Pearson Correlation Coefficient was applied. (G). Tissue prevalence estimated by Ro/e score. (H) Differentiation trajectory of DC subsets in CRC, with each color coded for pseudotime and clusters.
Figure 4 Immunological ACKR1+ DC were prevalent in left-sided CRC
(A) Violin plot showing the expression of ACKR1, SLC16A11, CD1A. (B) t-SNE plot showing the distribution of ACKR1+ DC. (C) Boxplot showing the proportions of ACKR1+ DC in left-sided and right-sided CRC. (D) Barplot showing the proportion of ACKR1+ DC across all DC subclusters. (E) Visualization the expression of ACKR1+ DC versus ACKR- DC markers. (E-J) Dotplot showing the correlation of ACKR1+ DC markers (ACKR1, SLC16A11, C1QA, CD1A, C1QC, EPHA8) and CD8. (K) Kaplan-Meier plot showing better clinical outcome in colon cancer with higher expression of ACKR1+ DC marker genes. (L) Bxoplot showing the AKCR1+ DC score over reponse (R) and non-response (NR) groups in TISMO colon cancer antiPD1 treatment assay. (M) Violin plot showing the DEGs between ACKR1+DC and ACKR1- DC cells. (N) Dotplot showing the DEPs between ACKR1+ DC and ACKR1- DC.
Figure S3 ACKR1+ DC is an prominent marker for prognosis
(A) t-SNE plot showing the distribution of ACKR1+ DC over left-sided and right-sided CRC. Violin plot showing the DEGs between ACKR1+ DC and ACKR1- DC in left-sided (B) and right-sided CRC (C). (D). Box plot showing the AKCR1+ DC score between Response and Non-response group in pancancer antiPD1 treatment assay, including lung cancer, melanoma, sarcoma, liver cancer, renal adenocarcinoma, and gstric cancer. Dot plot showing the DEPs between ACKR1+ DC and ACKR1- DC in left-sided (E) and right-sided CRC (F).
Figure 5 Monocytes from right-sided CRC exhibit higher level of granulocyte-migration-related genes
(A) t-SNE plot showing the distribution of monocyte subsets. (B) Circle plot showing the proportion of monocyte subsets between left-sided and right-sided CRC. (C) Dot plot showing the DEGs between left-sided and right-sided CRC in monocytes subsets. Top 5 genes were labelled. (D) Dot plot showing the GO enrichment terms between left-sided and right-sided CRC in monocyte subset. (E) Heatmap showing the correlations between monocytes in left-sided and right-sided CRC. (F) Co-expression of variance genes in left-sided and right-sided CRC. (G) Visualization of expression of granulocyte-migration-related genes between left-sided and right-sided CRC. (H) t-SNE plot showing the expression patterns of granulocyte-migration-related genes in monocytes.
Figure 6 Tissue-specific ligand-receptor pairs
Dot plot showing the left-sided-specific (A) and right-sided-specific (B-I) ligand-receptor pairs.