Resting natural killer cells promote the progress of colon cancer liver metastasis by elevating tumor-derived stem cell factor
- Department of General Surgery, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, China
- Department of Microbiology and Immunology, Institute of Molecular Virology and Immunology, School of Basic Medical Sciences, Wenzhou Medical University, China
- Department of Pediatric Thoracic Surgery, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, China
- Department of Thoracic Surgery, The First Affiliated Hospital of Wenzhou Medical University, China
- Department of General Surgery, The First Affiliated Hospital of Wenzhou Medical University, China
Figures
Figure 1 with 1 supplement
Landscape of tumor immune microenvironment in colon cancer liver metastasis (CCLM) revealed by single-cell transcriptomics.
(A) Schematic overview of the experimental design and analytical workflow. Colon T: colon tumor tissue; Colon P: colon paratumor normal tissue; Liver T: liver metastasis tumor tissue; Liver P: liver …
Figure 1—figure supplement 1
Cluster characterization of the global landscape of colon cancer liver metastasis (CCLM) revealed by single-cell transcriptomics.
(A) The UMAP plot of all 12 cell clusters. (B) Dot plots showing average expression of marker genes in indicated cell clusters. (C) The pie chart of the proportion of main immune cells across …
Figure 2 with 2 supplements
Cellular identification in spatial transcriptomic samples.
(A) UMAP visualization of cell clusters in spatial transcriptomic samples. (B) Dot plots showing average expression of markers in indicated cell clusters. (C) Overview of the spatial transcriptomic …
Figure 2—figure supplement 1
Cellular identification in spatial transcriptomic samples.
(A) The UMAP plot of all 11 cell clusters. (B) Dot plots showing average expression of marker genes in indicated cell clusters. (C) Expression of key markers across all samples.
Figure 2—figure supplement 2
Gene expression features of each sample.
The feature plots showed the expression distributions of ALB, CD24, COL1A1, MYH11, CXCL8, CD14, CD79A, and CD3D in each spatial transcriptomic section.
Figure 3
Clinical and biological relationship between natural killer (NK) cells and metastasis of colon cancer revealed by bulk RNA transcriptomics.
(A–D) The relationship of immune cell percentage determined by xCell, EPIC, quanTIseq, and MCPCounter between metastasis and non-metastasis tumor in TCGA COAD cohort. (*: p<0.05, **:p<0.01, ***：p<0.0…
Figure 4
The landscape of natural killer (NK) cells in the disease progression of colon cancer liver metastasis (CCLM).
(A) The UMAP plot of NK cells from CCLM. (B) Unsupervised clustering identifies eight subsets of NK cells. (C) Expression of key markers that distinguish resting and activated subsets of NK cells. (D…
Figure 5 with 1 supplement
Clinical relationship between natural killer (NK) cells subsets and metastasis of colon cancer revealed by bulk RNA and spatial transcriptomics.
(A, B) The relationship of activated and resting NK cell percentage determined by CIBERSORT and tumor metastasis in TCGA COAD cohort. M0: non-metastasis colon cancer; M1: metastasis colon cancer. …
Figure 5—figure supplement 1
Clinical relationship between natural killer (NK) cells subsets and colon cancer revealed by TCGA COAD cohort.
(A) The relationship of activated and resting NK cell and clinical characteristics of colon cancer in TCGA COAD cohort. (B) The relationship of 22 immune cells percentage determined by CIBERSORT and …
Figure 6 with 1 supplement
Characterization and developmental course of differential subsets of natural killer (NK) cells in colon cancer liver metastasis (CCLM).
(A) The UMAP plot of KIR2DL4+ activated NK cells, GZMK+ resting NK cells from CCLM. (B) Proportions of KIR2DL4+ activated NK cells, GZMK+ resting NK cells in total immune cells. p values were …
Figure 6—figure supplement 1
The proportions of eight clusters of NK cells in Colon P, Colon T, Liver T and LN.(*: p<0.05, **:p<0.01, ***：p<0.001).
Figure 7 with 5 supplements
Colon cancer cells (HCT-116) educated NK cells shift toward tumor-promoting status depends on cell-to-cell interaction.
(A, B) Clone formation assay showed the natural killer (NK) cell-mediated inductive effect on cell proliferation of colon cancer cell (HCT-116). Colon cancer cells were cultured in the supernatant …
Figure 7—figure supplement 1
Natural killer (NK) cell-mediated tumor-promoting effect in colon cancer cells (DLD-1).
(A) Clone formation assay showed the NK cell-mediated inductive effect on cell proliferation of DLD-1 cell between CNS and CS groups. (B, C) Cell Counting Kit-8 (CCK-8) assay showed the NK …
Figure 7—figure supplement 2
The positive gate locations of CD56, GZMK, KIR2DL4, CD9, CD49a, and PD-1 defined according to the Fluorescence Minus One (FMO) control.
Figure 7—figure supplement 3
Phenotype switch (CD56+, GZMK+) of natural killer (NK) cells was analyzed by fluorescence-activated cell sorting (FACS) after fixation and permeabilization in different co-cultured groups.
Figure 7—figure supplement 4
Phenotype switch of natural killer (NK) cells in different co-cultured system and the corresponding NK cell-mediated effect on cell migration of fresh colon cancer cell (HCT-116).
(A, B) NK cells underwent phenotype switch (high expression of CD9) when cocultured with human colon cancer cell line (HCT-116) and human fibroblast cell line (HFF-1), the phenotype switch was more …
Figure 7—figure supplement 5
Schematic overview of the in vitro experimental design.
(A) Schematic overview of coculture experiments design. (B) Schematic overview of coculture experiments design in transwells.
Figure 8 with 1 supplement
The resting natural killer (NK) cell promotes tumor malignant phenotype via elevating tumor-derived sSCF.
(A) Luminex liquid suspension chip detection of 48 common chemotactic and inflammatory cytokines in CNS, SNS, and MNS group. (B, C) Concentration of SCF determined by luminex liquid suspension chip …
Figure 8—figure supplement 1
Relative KITLG expression in different groups.
(A) Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) analysis of KITLG expression in HCT-116 with and without co-cultured with natural killer (NK) cells. (B) RT-qPCR …
Figure 9
Schematic diagram.
Colon cancer cells (HCT-116 cells) educate natural killer (NK) cells as resting status depends on cell-to-cell interaction. Tumor-educated NK cells subsequent enhances tumor malignancy in a …
Author response image 1
The positive gate locations of CD56, GZMK, KIR2DL4, CD9, CD49a, PD-1 defined according to the FMO control.
Author response image 2
Phenotype switch of NK cells in different co-cultured system and the corresponding NK cell-mediated effect on cell migration of fresh colon cancer cell (HCT-116).
A-B: NK cells underwent phenotype switch (high expression of CD9) when cocultured with HCT and HFF, the phenotype switch was more obvious when co-cultured with HCT. CN: NK cells cocultured with …
Author response image 3
Phenotype switch (CD56+, GZMK+) of NK cells was analyzed by FACS after fixation and permeabilization in different co-cultured groups.
CN: NK cells cocultured with colon cancer cells; SN: NK cells cocultured with supernatant of cancer cells; MN: NK cells cocultured in fresh medium.
Tables
Author response table 1
The baseline data of patient from single cell sequencing database.
| Patient | Age | Sex | TNM | Neoadjuvant chemotherapy |
|---|---|---|---|---|
| P1 | 71 | Male | T1N0M1 | N |
| P2 | 78 | Female | T3N1M1 | N |
| P3 | 60 | Female | T2N0M1 | N |
| P4 | 26 | Female | T3N1M1 | N |
| P5 | 64 | Male | T2N0M1 | Y (PR, FOLFOX) |
| P6 | 71 | Male | T3N1M1 | N |
| P7 | 52 | Female | T3N1M1 | Y (PR, XELOX) |
| P8 | 53 | Male | T3N0M1 | N |
| P9 | 35 | Male | T1N0M1 | Y (PR, XELOX) |
| P10 | 66 | Male | T3N1M1 | N |
| P11 | 43 | Female | T3N1M1 | N |
| P12 | 68 | Male | T3N1M1 | N |
| P13 | 69 | Female | T3N1M1 | Y (PD, XELOX) |
| P14 | 46 | Female | T3N1M1 | Y (PD, XELOX) |
| P15 | 51 | Female | T3N1M1 | Y (PR, FOLFOX) |
| P16 | 64 | Female | T3N1M1 | N |
| P17 | 49 | Female | T3N1M1 | N |
| P18 | 55 | Male | T3N1M1 | Y (PD, XELOX) |
| P19 | 63 | Female | T2N0M1 | Y (PD, XELOX) |
| P20 | 54 | Male | T3N0M1 | Y (SD, FOLFOX) |
Author response table 2
The baseline data of patient from spatial transcriptome database.
| Patient | Age | Sex | TNM | Treatment |
|---|---|---|---|---|
| ST-P1 | 73 | Male | T3N0M1 | N |
| ST-P2 | 67 | Female | T3N1M1 | N |
| ST-P3 | 72 | Male | T3N1M1 | Y (XELOX) |
| ST-P4 | 68 | Female | T3N1M1 | Y (XELOX) |
Additional files
-
Supplementary file 1
Characterized genes for Single-cell transcriptomic analysis.
- https://cdn.elifesciences.org/articles/97201/elife-97201-supp1-v1.docx
-
Supplementary file 2
Characterized genes for spatial transcriptomic analysis.
- https://cdn.elifesciences.org/articles/97201/elife-97201-supp2-v1.docx
-
Supplementary file 3
Characterized genes of natural killer (NK) subsets.
- https://cdn.elifesciences.org/articles/97201/elife-97201-supp3-v1.docx
-
Supplementary file 4
Characterized genes of natural killer NK of different status.
- https://cdn.elifesciences.org/articles/97201/elife-97201-supp4-v1.docx
-
Supplementary file 5
Fluorescence Minus One control of CD56, CD9, PD-1 and CD49a.
- https://cdn.elifesciences.org/articles/97201/elife-97201-supp5-v1.docx
-
Supplementary file 6
Fluorescence Minus One control of KIR2DL4 and GZMK.
- https://cdn.elifesciences.org/articles/97201/elife-97201-supp6-v1.docx
-
Supplementary file 7
Primers for quantitative real-time PCR (qPCR).
- https://cdn.elifesciences.org/articles/97201/elife-97201-supp7-v1.docx
-
MDAR checklist
- https://cdn.elifesciences.org/articles/97201/elife-97201-mdarchecklist1-v1.pdf
-
Source code 1
Source code used in this study.
- https://cdn.elifesciences.org/articles/97201/elife-97201-code1-v1.zip
