Article Figures & Data
Figures
- Figure 1.
Multiple myeloid cell subsets are present within tumors of different origin. A, analysis of myeloid cell subsets within subcutaneous 4T1 and Her2 tumors by flow cytometry. Hematopoietic cells were analyzed for the expression of CD11b and Gr1, and gated populations in the dot plots were further analyzed for CD11c expression. B, MHCII expression by indicated subsets. C, accumulation of CD11b+Gr1+ cells in the spleens of 4T1 tumor-bearing mice (top) and Gr1 expression (bottom). Numbers indicate percentage of cells for each gate or region. D, dendrogram analysis based on hierarchical clustering of tumor-associated and steady-state myeloid cells (BM, bone marrow; SI, small intestine; Spl, spleen; Tmr, tumor). Sample sizes are indicated in Materials and Methods. E, hierarchical clustering of the same populations based on a list of 139 genes associated with DCs, macrophages, or neutrophils.
- Figure 2.
Abundance of myeloid cell subsets across different tumors. A, Whisker box plots summarizing the relative frequency of myeloid subsets among hematopoietic cells within 4T1, Her2, and B16 tumors. B, frequency of CD3+ T cells within hematopoietic cells in 4T1, Her2, and B16 tumors. C, frequency of myeloid cells in 4T07, Pan02, EL4, CT26, PDA, RIP-Tag2, and A20 tumors (n = 3–7). *, P < 0.05; **, P < 001; ***, P < 0.001.
- Figure 3.
Tumor type dictates the composition of myeloid cell subsets within tumors. A, myeloid cells within subcutaneous and mammary gland of transgenic Her2 tumors (n = 4–7); within primary subcutaneous and spontaneous peritoneal metastatic 4T1 tumors (n = 3); and within subcutaneous B16 tumors or lung and kidney metastases (n = 3–4). B, left, analysis of myeloid subsets within tumors from Balb/c mice with single (Her2 or 4T1; n = 5) or double (Her2 and 4T1 on opposite flanks; n = 8) tumors. Right, frequency of myeloid cells in the spleens of tumor-bearing mice. C, left, analysis of myeloid cell subsets within tumors from F1 (Balb/c × C57BL/6) mice with single (Her2 or B16; n = 3) or double (Her2 and B16 on opposite flanks; n = 3) tumors. Right, frequency of myeloid cells in the spleens of tumor-bearing mice. Numbers indicate percentage of cells for each gate or region. N.S., not statistically significant; *, P < 0.05; ***, P < 0.001.
- Figure 4.
Tumor-infiltrating myeloid cells express immunomodulatory genes. A, comparison of gene expression profiles of CD11c+ and CD11c− TAMs to Gr1low MDSCs from 4T1 tumor-bearing mice based on 2-fold change (fold-change vs. fold-change plot). B, comparison of gene expression profiles of TANs and Gr1hi MDSCs from 4T1 tumor-bearing mice (fold-change vs. t test P value plot). C, heatmap generated by hierarchical clustering using probes identified in A and B for all tumor myeloid subsets. D, heatmap showing the expression of hypoxia-related genes by tumor-associated and steady-state myeloid cells. Selected genes associated with the populations are indicated on the plots.
- Figure 5.
Expression of protumorigenic genes by TANs. A, comparison of gene expression profiles of TANs and Gr1hi MDSCs from 4T1 tumor-bearing mice to naïve neutrophils (left, fold-change vs. fold-change, 2-fold change). Expression of these probes was further analyzed in fold-change versus fold-change plots comparing TANs from B16 and Her2 tumors with naïve neutrophils (right); 170 of the 214 probes passed the CV < 0.5 filter. B, heatmap showing the expression of the 170 probes identified in A across neutrophils. C, top 20 canonical pathways enriched for TANs and Gr1hi MDSCs compared with naïve neutrophils analyzed by Ingenuity Pathway Analysis. ROS, reactive oxygen species.
- Figure 6.
Tumor type–specific transcriptional profiles of TANs. A, fold-change versus fold-change plot comparing 4T1 TANs with TANs in Her2 and B16, based on 2-fold change. Selected genes associated with the populations are indicated on the plots. B, chemokine, chemokine receptor, cytokine, and cytokine receptor expression profiles of TANs. C, PPARα-associated gene expression profiles of neutrophils. D, principal component analysis of neutrophil populations.
- Figure 7.
Altered expression profile of secretory granule molecules in TANs. A, expression of granular effector molecules by neutrophils. B, mean expression value of haptoglobin (Hp, HP) across neutrophils and tumor-associated myeloid cells. C, serum haptoglobin levels in mice with small (S), medium (M), large (L) tumors and naïve mice. D, correlation between the frequency of Gr1hi cells in blood and the serum haptoglobin concentration. E, haptoglobin levels in serum from healthy donors (n = 4) and patients with breast cancer (n = 10). *, P < 0.05; **, P < 0.01.
Additional Files
Supplementary Data
Files in this Data Supplement:
- Supplementary Figure Legends - PDF file - 63K
- Supplementary Figure 1 - PDF file - 136K, Supplementary Figure S1: Additional figures on myeloid cell characterization in different tumor models.
- Supplementary Figure 2 - PDF file - 42K, Supplementary Figure S2: Additional figures on myeloid cell characterization based on tumor type and anatomical location.
- Supplementary Table 3 - XLSX file - 12K, Supplementary Table S3: Expression values for neutrophil-associated genes from the heatmap in Figure 5B.
- Supplementary Table 1 - XLSX file - 29K, Supplementary Table S1: Expression values for myeloid cell signature genes from the heatmap in Figure 1E.
- Supplementary Table 6 - XLSX file - 1K, Supplementary Table S6: Expression values for neutrophil granular effector molecules from the heatmap in Figure 7A.
- Supplementary Table 2 - XLSX file - 7K, Supplementary Table S2: Expression values for hypoxia-related genes from the heatmap in Figure 4D.
- Supplementary Table 5 - XLSX file - 2K, Supplementary Table S5: Expression values for PPARa-associated genes from the heatmap in Figure 6C.
- Supplementary Table 4 - XLSX file - 4K, Supplementary Table S4: Expression values for neutrophil cytokine-chemokine genes from the heatmap in Figure 6B.
Volume 2, Issue 7
