pdac cell lines aspc 1  (ATCC)

Journal: Nature Communications

Article Title: Targeting TNK2/ACK1 reverses the immunosuppressive tumor microenvironment and synergizes with immunochemotherapy in pancreatic cancer

doi: 10.1038/s41467-025-67197-3

Figure Lengend Snippet: a Schematic illustration of the screening strategy created created in BioRender. wu, c. (2025) https://BioRender.com/9r8yhl5 . b Representative spatial partition showing “Cold_desert” zones and “Hot_inflammatory” zones defined by CD8 + T cell distribution; 120 spots were randomly sampled from “Cold_desert” and 160 from “Hot_inflammatory” across 10 cases ( GSM3036911 ). Each spot diameter 50 µm. Differential expression genes between the two spot groups are provided as DEGs1 in Supplementary Data . c Representative images grouping into “Cold_desert” samples and “Hot_inflammatory” samples based on CD8 + T cell infiltration across the same 10 cases ( GSM3036911 ); differential expression genes between two groups are provided as DEGs2 in Supplementary Data . d Representative mIHC images from 13 cases (HRA000433) illustrating “Cold_desert” tumors and “Hot_inflammatory” tumors. And 100 areas were randomly sampled from “Cold_desert” tumors and 80 from “Hot_inflammatory” tumors for CD8 + T cell fraction quantification; each area diameter 100 µm. Differential expression genes between two groups are provided as DEGs3 in Supplementary Data . CD8 + T cells: red, CK19: cyan, nucleus: blue. e DEG4 denotes tumor versus normal differential expression genes from TCGA-PAAD and GTEx that are positively associated with the immunosuppressive state within TCGA tumors (provided in Supplementary Data ). f Venn diagram showing the intersection used to derive genes positively associated with the immunosuppressive tumor microenvironment in PDAC. Four differential gene sets were intersected. Selection thresholds were: DEGs1 log2FC ≥ 1.5, p < 0.05; DEGs2 log2FC ≥ 1.3, p < 0.05; DEGs3 log2FC ≥ 1.5, p < 0.05; DEGs4 log2FC ≥ 1.0, p < 0.05. g The representative images of TNK2 IHC staining at different expression levels ( n = 70 independent samples). h The heatmap of the differential expression of TNK2 in 70 pairs of tumor tissues (T) and corresponding adjacent non-tumor tissues (N). i , j Survival analysis of PDAC patients with low and high TNK2 expression based on the dataset of TJMUCH, and p value from log-rank test are shown. k Statistical analysis of OS, RFS and PFS based on a multi-center retrospective cohort for TNK2 Hazard ratio ( GSE78229 : n = 49; GSE62452 : n = 65; GSE71729 : n = 125; ICGC-PAAD-AU seq: n = 81; TCGA-PAAD: n = 150; ICGC-PAAD-AU-array: n = 103; E-MTAB-6134: n = 288; ICGC-PAAD-CA-seq: n = 113. All n represents independent samples). Hazard ratio with 95% confidence intervals, box shows the interquartile range (IQR), line indicates the median, and whiskers the min/max values; tests two sided; exact p values are reported in the legends. Scale bars:100 µm. Source data are provided as a Source Data file.

Article Snippet: The human PDAC cell lines AsPC-1, MIA PaCa-2, Panc 10.05, Capan-2, CFPAC-1 BxPC-3 and SW1990, and human embryonic kidney cell line HEK-293T, were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Quantitative Proteomics, Selection, Immunohistochemistry, Expressing

Journal: Nature Communications

Article Title: Targeting TNK2/ACK1 reverses the immunosuppressive tumor microenvironment and synergizes with immunochemotherapy in pancreatic cancer

doi: 10.1038/s41467-025-67197-3

Figure Lengend Snippet: a – c Interactions of the TNK2_high Malignant ductal cells. a TNK2_low Malignant ductal cells ( b ) and CD8 + T cells ( c ) with other cell types indicated in the figure, as determined by cell-cell contact analysis. d Representative images of the mIHC staining of PDAC tissue microarray ( n = 98 independent samples) with high (left) or low (right) TNK2 expression. Images are representative of results observed across the cohort. CD8 + T cells: red, NK cells (NKp60 + cells): orange, MDSC cells (CD33 + cells): cyan, Tregs (Foxp3 + cells): yellow, CK19 + cells: pink, and nuclei: blue. Scale bars: 50 μm. e Correlation between counts of TNK2 + CK19 + cells and counts of CD8 + T cells in PDAC samples ( n = 98 independent samples). The r value was determined based on the spearman correlation coefficient, two-sided p value is reported. f Correlation between tumor-cell TNK2 expression and the percentage of CD8⁺ T cells was assessed using the single-cell RNA-seq dataset CRA001160 ( n = 24 independent primary PDAC tumors). Tumors were stratified by the median tumor-cell TNK2 expression into TNK2-High and TNK2-Low groups (12 cases each). g The representative mIHC staining images of PDAC tissue microarray (left) ( n = 98 independent samples) with TNK2 in green, CK19 in cyan, CD8 + T cell in red and nuclei in blue, the closest spatial distance analysis (right) with TNK2 + CK19 + cells in yellow dots and CD8 + T cells in blue dots, the lines were the connections between them. Scale bars: 50 μm. The statistical analysis of the number of CD8 + T cells ( h ) and the nearest distance between TNK2 + CK19 + cells and CD8 + T cells ( i ) in PDAC samples with different TNK2 expression. Analysis of the TNK2 expression from 33 randomly selected regions on 10 spatial transcriptome sequencing samples ( j ) and the average distance of the nearest CD8 + T cells in these selected regions ( k ). Two-sided Welch t test with Benjamini–Hochberg correction; adjusted p values are indicated ( k ). Scale bars: 50 μm. Statistical significance was determined by two-sided Wilcoxon rank-sum tests ( f , h , i ), box shows the interquartile range (IQR), line indicates the median, and whiskers the min/max values. Source data are provided as a Source Data file.

Article Snippet: The human PDAC cell lines AsPC-1, MIA PaCa-2, Panc 10.05, Capan-2, CFPAC-1 BxPC-3 and SW1990, and human embryonic kidney cell line HEK-293T, were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Staining, Microarray, Expressing, RNA Sequencing, Sequencing

Journal: Nature Communications

Article Title: Targeting TNK2/ACK1 reverses the immunosuppressive tumor microenvironment and synergizes with immunochemotherapy in pancreatic cancer

doi: 10.1038/s41467-025-67197-3

Figure Lengend Snippet: a Schematic illustration of the co-culture experiment for activated CD8 + T cells and KPC cells created in BioRender. wu, c. (2025) https://BioRender.com/9r8yhl5 . The flow cytometry analysis showing the changes of TNFα + CD8 + T cells ( b ), PD-1 + CD8 + T cells ( c ) and proliferative CD8 + T cells ( d ) in the co-culture system ( n = 3 independent experiments). e Schematic illustration of the co-culture experiment for activated OT-I-td-Tomato CD8 + T cell and KPC cells created in BioRender. wu, c. (2025) https://BioRender.com/9r8yhl5 . f Percentage of apoptotic KPC#3-OVA or KPC#1-OVA cell lines after co-cultured with activated OT-I-td-Tomato CD8 + T cells, as measured by flow cytometry after staining with PI and annexin-V ( n = 3 independent experiments). The representative fluorescence images of KPC#3-OVA or KPC#1-OVA cell lines stained with caspase-3/7 probes (green) which cocultured with OT-I-td-Tomato CD8 + T cell (red) ( g , h ) and fluorescence intensity quantified with ImageJ ( i ) ( n = 3 independent experiments). j The LDH release assay to determine the cytotoxicity of OT-I-td-Tomato CD8 + T cells. k Schematic illustration of the apoptosis assay in PDAC organoid created in BioRender. wu, c. (2025) https://BioRender.com/9r8yhl5 . The representative fluorescence images of PDAC#5 or PDAC#4 organoids stained with caspase-3/7 probes (green) which co-cultured with activated PBMC ( l ), and quantified by ImageJ ( m ) ( n = 3 independent experiments). Scale bars, 100 μm. Statistical significance was determined by two-tailed unpaired Student’s t test. Data represent means ± s.d. Source data are provided as a Source Data file.

Article Snippet: The human PDAC cell lines AsPC-1, MIA PaCa-2, Panc 10.05, Capan-2, CFPAC-1 BxPC-3 and SW1990, and human embryonic kidney cell line HEK-293T, were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Co-Culture Assay, Flow Cytometry, Cell Culture, Staining, Fluorescence, Lactate Dehydrogenase Assay, Apoptosis Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Targeting TNK2/ACK1 reverses the immunosuppressive tumor microenvironment and synergizes with immunochemotherapy in pancreatic cancer

doi: 10.1038/s41467-025-67197-3

Figure Lengend Snippet: a Venn diagram illustrating the screening strategy for TNK2-regulated cell surface proteins. Candidate genes were obtained from the intersection of four datasets: immune-regulatory proteins, known membrane proteins, proteins with significant membrane peptides identified by surface proteomics after TNK2 overexpression, and genes upregulated in PDAC with fold change ≥2. Six overlapping genes (HVEM, PPIA, GPI, PSMC2, CALR, and RAC1) were identified. b ScRNA-seq data analysis, UMAP plot were annotated and colored for different cell types (left panel). Expression levels of TNK2 and HVEM were lighted in UMAP individually (2 middle panels) or together (right panel). c Hierarchical plot showing the inferred intercellular communication network for HVEM signaling. Solid and open circles represent source and target, respectively. Circle edge width represents the communication probability. Edge colors are consistent with the signaling source. d Heatmap showing the relative importance of each cell group based on the computed four network centrality measures (Sender, Receiver, Mediator, Influencer) of HVEM signaling pathway network. Representative pancreatic tumor images ( e , j ) and tumor size measured at the experimental endpoint ( f , k ) in tumors formed by KPC#3 cells expressing TNK2 cDNA or vector ( e , f ) or KPC#1 cells expressing TNK2 shRNA or control ( j – k ). Flow cytometry analysis showing the changes of CD8 + T cells ( g , l ), TNFα + CD8 + T cells ( h , m ), and PD-1 + CD8 + T ( i , n ) cells in tumors formed by KPC#3 cells expressing TNK2 cDNA or vector ( g – i ) or KPC#1 cells expressing TNK2 shRNA or control ( l – n ). Statistical significance was determined by two-way ANOVA followed by Tukey’s multiple comparisons test ( f , k ) and one-way ANOVA followed by Tukey’s multiple comparisons test ( g – i , l – n ). n = 8 independent experiments ( e – n ). All tests were two sided. Data represent as mean ± s.d. Source data are provided as a Source Data file.

Article Snippet: The human PDAC cell lines AsPC-1, MIA PaCa-2, Panc 10.05, Capan-2, CFPAC-1 BxPC-3 and SW1990, and human embryonic kidney cell line HEK-293T, were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Membrane, Over Expression, Expressing, Plasmid Preparation, shRNA, Control, Flow Cytometry

Journal: Nature Communications

Article Title: Targeting TNK2/ACK1 reverses the immunosuppressive tumor microenvironment and synergizes with immunochemotherapy in pancreatic cancer

doi: 10.1038/s41467-025-67197-3

Figure Lengend Snippet: a Representative images of TNK2 IHC staining and CT scans in PDAC patients treated with neoadjuvant therapy. b Association between TNK2 expression level and neoadjuvant chemotherapy response. Counts of PR, SD and PD are shown for TNK2-low ( n = 22) and TNK2-high ( n = 23) tumors (total n = 45 independent experiments). The overall association across the three response categories was tested by Pearson’s χ² test (two-sided); p = 0.0046. For the binary summary, NPD = PR + SD versus PD (comparison indicated in the panel). PR partial remission, SD stable disease, PD progressive disease, NPD non-progressive disease. c The synergy score of GEM, AIM100 and αPD-1 were analyzed through Synergy finder for the treatment of orthotopic transplantation mouse model. The representative tumor images ( d up) and the representative PET-CT images ( d down, e ) of the KPC GEMM mouse which treated with control, AG, AIM100 and AG, AG and aPD-1, or AG, AIM100 and αPD-1, at the experimental endpoint before the therapy ending stage ( n = 3 independent experiments). f The pancreas weights of the mouse groups described in ( d ) ( n = 3 independent experiments). g Kaplan–Meier curves for tumor free survival in the mouse groups in ( d ). Flow cytometry analysis showing the proportions of CD8 + T cells ( h ), TNFα + CD8 + T cells ( i ), and PD-1 + CD8 + T ( j ) cells in the mouse groups in ( d ). k The representative mIHC images of tumor issues (CK19 in cyan, Ki67 in red, and nuclei in blue) from the mouse groups in ( d ). l The representative mIHC images of tumor issues from the mouse groups in ( d ) at the experimental endpoint, with TNK2 stained in blue, CD8 in red, CK19 in purple, and nuclei in blue. Statistical significance was determined by log-rank (Mantel–Cox) test in survival analysis ( g ) or one-way ANOVA followed by Tukey’s multiple comparisons test ( e , f , h – l ). All tests were two sided ( e – l ). Data represent mean ± s.d. Source data are provided as a Source Data file.

Article Snippet: The human PDAC cell lines AsPC-1, MIA PaCa-2, Panc 10.05, Capan-2, CFPAC-1 BxPC-3 and SW1990, and human embryonic kidney cell line HEK-293T, were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Immunohistochemistry, Expressing, Comparison, Transplantation Assay, Positron Emission Tomography-Computed Tomography, Control, Flow Cytometry, Staining