pancreatic ductal adenocarcinoma pdac cell lines panc 1  (ATCC)

Journal: Cancer Research

Article Title: Radiation-Enhanced CD24 Membrane Trafficking via GPI Anchoring Mediates Antitumor Immune Evasion

doi: 10.1158/0008-5472.CAN-25-2616

Figure Lengend Snippet: Radiation fails to enhance macrophage phagocytosis despite inducing immunogenic signals. A, Schematic illustration of scRNA-seq in animal models. t-SNE, t-distributed stochastic neighbor embedding. B, scRNA-seq analysis of immunogenic markers on tumor cells from control (Ctrl) or irradiated LLC tumors ( n = 3). C and D, Flow cytometry histograms and mean fluorescence intensity (MFI) of CRT on control or irradiated H460 ( C ) and PANC-1 ( D ) cells ( n = 3). Normalized to the control group. E, Immunofluorescence images of CRT (red) on control or irradiated H460 cells. F, Flow cytometry histograms and mean fluorescence intensity of tumor cells CRT from control or irradiated KPC tumor models ( n = 6). Normalized to the control group. G and H, Culture medium HMGB1 levels from control or irradiated H460 ( G ) and PANC-1 ( H ) cells measured by ELISA ( n = 3). I and J, Extracellular ATP release from control or irradiated H460 ( I ) and PANC-1 ( J ) cells measured by ELISA ( n = 3). K, scRNA-seq–derived M1 signature scores of tumor-associated macrophages from irradiated or control LLC tumors ( n = 3). L, Flow cytometry histograms and quantitation of control or irradiated H460 cell phagocytosis by THP-1 macrophages ( n = 3). M, Immunofluorescence images and quantitation of control or irradiated H460 cell (green) phagocytosis by THP-1 macrophages (red), with arrows indicating phagocytic events ( n = 6). Scale bars, 50 μm. N, Flow cytometry dotplot and quantitation of tumor cell phagocytosis by tumor-associated macrophages from control or irradiated LLC tumor ( n = 6). IR, 8 Gy irradiation. All data are presented as mean ± SEM and compared using a two-tailed Student t test. NS, not significant. A, Created in BioRender. Kong, L. (2026) https://BioRender.com/rwdzfwn .

Article Snippet: The human pancreatic ductal adenocarcinoma (PDAC) cell lines PANC-1 (RRID: CVCL_0480) and BxPC-3 (RRID: CVCL_0186), human non–small cell lung cancer cell lines NCI-H460 (RRID: CVCL_0459) and NCI-H1299 (RRID: CVCL_0060), human monocytic cell line THP-1 (RRID: CVCL_0006), murine hepatocellular carcinoma cell line Hepa1-6 (RRID: CVCL_0327), and murine lung carcinoma cell line Lewis (RRID: CVCL_4358) were obtained from the ATCC.

Techniques: Control, Irradiation, Flow Cytometry, Fluorescence, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Derivative Assay, Quantitation Assay, Two Tailed Test

Journal: Cancer Research

Article Title: Radiation-Enhanced CD24 Membrane Trafficking via GPI Anchoring Mediates Antitumor Immune Evasion

doi: 10.1158/0008-5472.CAN-25-2616

Figure Lengend Snippet: Radiation enhances tumor cell “do not eat me” signal CD24. A, UMAP plots of pancreatic adenocarcinoma with 10 clusters, NCBI Sequence Read Archive: GSE281288 . CD24 expression overlaid onto UMAP. B, Immunofluorescence images of CD24 (red) on control or irradiated H460 cells. Scale bars, 25 and 5 µm. C and D, Flow cytometry histograms and mean fluorescence intensity (MFI) of CD24 surface expression on control or irradiated H460 ( C ) and PANC-1 ( D ) cells ( n = 3). E, Tumor cell CD24 mean fluorescence intensity from control or irradiated KPC tumor models ( n = 6). F, Flow cytometry histograms and mean fluorescence intensity of CD24 surface expression on H460 cells 48 hours after 2–18 Gy of radiation ( n = 3). IR, 8 Gy irradiation. All data are presented as mean ± SEM and normalized to the control group. A two-tailed Student t test was performed for C–E . One-way ANOVA with a Tukey multiple comparisons test was performed for F . NS, not significant.

Article Snippet: The human pancreatic ductal adenocarcinoma (PDAC) cell lines PANC-1 (RRID: CVCL_0480) and BxPC-3 (RRID: CVCL_0186), human non–small cell lung cancer cell lines NCI-H460 (RRID: CVCL_0459) and NCI-H1299 (RRID: CVCL_0060), human monocytic cell line THP-1 (RRID: CVCL_0006), murine hepatocellular carcinoma cell line Hepa1-6 (RRID: CVCL_0327), and murine lung carcinoma cell line Lewis (RRID: CVCL_4358) were obtained from the ATCC.

Techniques: Sequencing, Expressing, Immunofluorescence, Control, Irradiation, Flow Cytometry, Fluorescence, Two Tailed Test

Journal: Cancer Research

Article Title: Radiation-Enhanced CD24 Membrane Trafficking via GPI Anchoring Mediates Antitumor Immune Evasion

doi: 10.1158/0008-5472.CAN-25-2616

Figure Lengend Snippet: CD24 inhibition combined with radiation promotes macrophages phagocytosis and activation. A and B, Flow cytometry histograms of CD24 surface expression on H460 ( A ) and PANC-1 ( B ) cells transfected with CD24 -targeting siRNA (si CD24 ), negative-control siRNA (siNC), or isotype control. C and D, Flow cytometry histograms and quantitation of H460 ( C ) and PANC-1 ( D ) cell phagocytosis by THP-1 macrophages, with the indicated treatments ( n = 3). E, Immunofluorescence images and quantitation of H460 cell phagocytosis by THP-1 macrophages (red), with the indicated treatments (green). Arrows, phagocytic events ( n = 6). Scale bars, 50 μm. F, qRT-PCR analysis of Siglec- 10 in THP-1 macrophages transfected with si Siglec- 10 or siNC ( n = 3). G, Flow cytometry histograms and quantitation of irradiated H460 cell phagocytosis by THP-1 macrophages, with the indicated treatments ( n = 3). H – J, Flow cytometry histograms and mean fluorescence intensity (MFI) of CD80 ( H ), CD86 ( I ), and PD-L1 ( J ) on BMDMs cocultured with tumor cells, with the indicated treatments ( n = 3). IR, 8 Gy irradiation. All data are presented as mean ± SEM. One-way ANOVA with a Tukey multiple comparisons test was performed for C–E and G–J . A two-tailed Student t test was performed for F . NS, not significant.

Article Snippet: The human pancreatic ductal adenocarcinoma (PDAC) cell lines PANC-1 (RRID: CVCL_0480) and BxPC-3 (RRID: CVCL_0186), human non–small cell lung cancer cell lines NCI-H460 (RRID: CVCL_0459) and NCI-H1299 (RRID: CVCL_0060), human monocytic cell line THP-1 (RRID: CVCL_0006), murine hepatocellular carcinoma cell line Hepa1-6 (RRID: CVCL_0327), and murine lung carcinoma cell line Lewis (RRID: CVCL_4358) were obtained from the ATCC.

Techniques: Inhibition, Activation Assay, Flow Cytometry, Expressing, Transfection, Negative Control, Control, Quantitation Assay, Immunofluorescence, Quantitative RT-PCR, Irradiation, Fluorescence, Two Tailed Test

Journal: Cancer Research

Article Title: Radiation-Enhanced CD24 Membrane Trafficking via GPI Anchoring Mediates Antitumor Immune Evasion

doi: 10.1158/0008-5472.CAN-25-2616

Figure Lengend Snippet: Radiation enhances CD24 membrane trafficking via GPI anchoring. A and B, Western blot analysis of CD24 in control or irradiated H460 ( A ) and PANC-1 ( B ) whole-cell lysate. C, Schematic representation of GPI anchoring. D, Western blot analysis of CD24 in control or irradiated H460 cell membrane and cytosolic lysate. E, Proteomic analysis heatmap of differential proteins in control or irradiated H1299. F and G, Western blot analysis of GPAA1 in control or irradiated H460 ( F ) and PANC-1 ( G ) whole-cell lysate. H, Western blot analysis of GPAA1 and CD24 in H460 cells transfected with si GPAA1 or siNC. I, Flow cytometry histograms of CD24 surface expression on H460 cells, with the indicated treatment or isotype control ( n = 3). MFI, mean fluorescence intensity. J, Flow cytometry histograms and quantitation of H460 cell phagocytosis by THP-1 macrophages, with the indicated treatments ( n = 3). IR, 8 Gy irradiation. All data are presented as mean ± SEM and compared using one-way ANOVA with a Tukey multiple comparisons test. NS, not significant.

Article Snippet: The human pancreatic ductal adenocarcinoma (PDAC) cell lines PANC-1 (RRID: CVCL_0480) and BxPC-3 (RRID: CVCL_0186), human non–small cell lung cancer cell lines NCI-H460 (RRID: CVCL_0459) and NCI-H1299 (RRID: CVCL_0060), human monocytic cell line THP-1 (RRID: CVCL_0006), murine hepatocellular carcinoma cell line Hepa1-6 (RRID: CVCL_0327), and murine lung carcinoma cell line Lewis (RRID: CVCL_4358) were obtained from the ATCC.

Techniques: Membrane, Western Blot, Control, Irradiation, Transfection, Flow Cytometry, Expressing, Fluorescence, Quantitation Assay

Journal: Nature Communications

Article Title: T cells engineered against Dickkopf-1-A2 complex can be used to treat HLA-A2 + solid and hematologic cancers

doi: 10.1038/s41467-026-69621-8

Figure Lengend Snippet: a Expression of DKK1 mRNA across hematologic tumors was ranked, and datasets of GSE13591 , GSE2350 , and GSE13159 from Oncomine database were used. MM (multiple myeloma), Pro-B-ALL (Pro-B cell acute lymphoblastic leukemia), MGUS (monoclonal gammopathy of undetermined significance), PCL (primary plasma cell leukemia), B-ALL Child (B-Cell acute lymphoblastic leukemia in children), FL (follicular lymphoma), B-ALL (B-cell acute lymphoblastic leukemia), MS (myelodysplastic syndrome), CML (chronic myeloid leukemia). b Expression of DKK1 mRNA across TCGA solid tumors compared to paired normal tissues was ranked in GEPIA2 database. PAAD (pancreatic adenocarcinoma), CHOL (cholangiocarcinoma), ESCA (esophageal carcinoma), HNSC (head and neck squamous cell carcinoma), LUSC (lung squamous cell carcinoma), LIHC (liver hepatocellular carcinoma), STAD (stomach adenocarcinoma), THYM (thymoma), UCS (uterine carcinosarcoma), KIRC (kidney renal clear cell carcinoma), LUAD (lung adenocarcinoma), ACC (adrenocortical carcinoma), CESC (cervical squamous cell carcinoma and endocervical adenocarcinoma). c , d Overall and disease-free survival of pancreatic cancer patients with DKK1 -high versus DKK1 -low expression in GEPIA2 database. The top 50% patients with higher expressions of DKK1 were defined as DKK1 -high, and the rest were defined as DKK1 -low patients. Hazards ratio and p value are demonstrated. Cox proportional hazards regression was used. e Solid tumor datasets of Pei Pancreas ( GSE16515 , normal n = 16, tumor n = 36), Landi Lung ( GSE10072 , normal n = 49, tumor n = 58) and Curtis Breast (EGAS00000000083, normal n = 144, tumor n = 250) from Oncomine database showing the expression of DKK1 mRNA in tumor and paired normal tissues. Two-tailed Student’s t -test was used. f Immunohistochemistry staining of DKK1-A2 complex on normal pancreatic, lung, or breast tissues from 5 randomly selected HLA-A2 positive donors. g–i Immunohistochemistry staining of DKK1, HLA-A2, and DKK1-A2 complex of 10 randomly selected HLA-A2 positive or negative PDAC, NSCLC, and TNBC tumor tissues. The independent experiments were repeated three times, and representative images are showed for immunohistochemistry staining. Data shown as mean ± SEM. Source data are provided as a file.

Article Snippet: Human mantle cell lymphoma cell line JeKo-1, Burkitt’s lymphoma cell line Daudi, MM cell lines U266, KMS-26, ARP-1, KMS-11, KMS-12-BM, and MM.1S, PDAC cell lines CFPAC-1, PANC-1, MIA PaCa-2, and PL45, NSCLC cell lines NCI-H441 and NCI-H266, and TNBC cell lines MDA-MB-231 and MDA-MB-468 were purchased from American Type Culture Collection or obtained from Arkansas Cancer Research Center.

Techniques: Expressing, Clinical Proteomics, Two Tailed Test, Immunohistochemistry, Staining

Journal: Nature Communications

Article Title: T cells engineered against Dickkopf-1-A2 complex can be used to treat HLA-A2 + solid and hematologic cancers

doi: 10.1038/s41467-026-69621-8

Figure Lengend Snippet: a Schematic presentation showing the recognition of DKK1-A2 CAR-T cells and DKK1-A2 expressing tumor cells. b Schematic presentation showing DKK1-A2 and CD19 CAR constructs. c Percentage of CAR expressing T cells determined as eGFP positive T cells by flow cytometry analysis. d Expression of DKK1, HLA-A2 or DKK1-A2 complex by JeKo-1, Daudi, U266, KMS-26, CFPAC-1, PANC-1, NCI-H441 or MDA-MB-231 tumor cell lines determined by flow cytometry. e CAR-T cell proliferation measured by dilution of Celltrace Violet analyzed by flow cytometry. CAR-T cells were stained with Celltrace Violet and co-cultured with JeKo-1, Daudi, U266 or CFPAC-1 tumor cells at an E:T ratio of 1:1 for 72 h. f Cytotoxicity of CAR-T cells determined by luciferase reporter assay. CAR-T cells were co-cultured with Luc-transduced JeKo-1, Daudi, U266, KMS-26, CFPAC-1, PANC-1, NCI-H441, or MDA-MB-231 tumor cells at indicated E:T ratios for 12 h. n = 3, technical replicates. g , h Percentages of IFN-γ and TNF-α producing CD4 + or CD8 + CAR-T cells determined by intracellular cytokine staining. CAR-T cells were co-cultured with JeKo-1, Daudi, MM.1S-A2, KMS-12-BM-A2, CFPAC-1 or PANC-1 tumor cells at an E:T ratio of 1:1 overnight. In the experiments, CD19 CAR-T cells were used as a control. n = 3, technical replicates. Data shown as mean ± SEM. Source data are provided as a file.

Article Snippet: Human mantle cell lymphoma cell line JeKo-1, Burkitt’s lymphoma cell line Daudi, MM cell lines U266, KMS-26, ARP-1, KMS-11, KMS-12-BM, and MM.1S, PDAC cell lines CFPAC-1, PANC-1, MIA PaCa-2, and PL45, NSCLC cell lines NCI-H441 and NCI-H266, and TNBC cell lines MDA-MB-231 and MDA-MB-468 were purchased from American Type Culture Collection or obtained from Arkansas Cancer Research Center.

Techniques: Expressing, Construct, Flow Cytometry, Staining, Cell Culture, Luciferase, Reporter Assay, Control

Journal: Nature Communications

Article Title: T cells engineered against Dickkopf-1-A2 complex can be used to treat HLA-A2 + solid and hematologic cancers

doi: 10.1038/s41467-026-69621-8

Figure Lengend Snippet: a Schema of U266 MM xenograft model treated with CAR-T cells, b Tumor burden measured as the levels of human Igλ secreted by U266 cells. Blood was collected from U266 xenografted mice weekly and human Igλ in the serum was measured by ELISA, c Survival curve of U266 xenografted mice monitored daily before and post CAR-T cell infusion, and d Body weight change of U266 xenografted mice. Ctrl, n = 4; CD19 CAR-T, n = 5; mC2 CAR-T, n = 5; biological replicates. e Schema of CFPAC-1-Luc PDAC orthotopic xenograft model treated with CAR-T cells, f Tumor burden shown as bioluminescence intensity of CFPAC-1-Luc PDAC orthotopic xenograft mice, g Survival curve of CFPAC-1-Luc PDAC orthotopically xenografted mice monitored daily before and post CAR-T cell infusion, h Body weight change of CFPAC-1-Luc PDAC orthotopic xenograft mice, and i Bioluminescence images of CFPAC-1-Luc PDAC orthotopic xenograft mice before and post CAR-T cell infusion. Ctrl, n = 5; mC2 CAR-T, n = 5; biological replicates. j Schema of NCI-H441-Luc NSCLC metastatic xenograft model treated with CAR-T cells infusion, k Bioluminescence intensity of NCI-H441-Luc NSCLC metastatic xenograft mice, l Survival of NCI-H441-Luc NSCLC metastatic xenograft mice, m Body weight change of NCI-H441-Luc NSCLC metastatic xenograft mice, and n Bioluminescence images of NCI-H441-Luc NSCLC metastatic xenograft mice before and post CAR-T cell infusion. Ctrl, n = 4; CD19 CAR-T, n = 4; mC2 CAR-T, n = 5; biological replicates. o Schema of MDA-MB-231-Luc TNBC orthotopic xenograft model treated with CAR-T cells, p Bioluminescence intensity of MDA-MB-231-Luc TNBC orthotopic xenograft mice, q Survival of MDA-MB-231-Luc TNBC orthotopic xenograft mice, r Body weight change of MDA-MB-231-Luc TNBC orthotopically xenografted mice, and s Bioluminescence images of MDA-MB-231-Luc TNBC orthotopic xenograft mice before and post-CAR-T cell infusion. Ctrl, n = 5; CD19 CAR-T, n = 5; mC2 CAR-T, n = 5; biological replicates. Two-way ANOVA was used for bioluminescence, tumor volume, and body weight analyses (Two-tailed Student’s t -test for CFPAC-1-Luc PDAC orthotopic xenograft model). Log-rank test was used for survival analysis. In most experiments, CD19 CAR-T cells were used as a control. Data shown as mean ± SEM. Source data are provided as a file.

Article Snippet: Human mantle cell lymphoma cell line JeKo-1, Burkitt’s lymphoma cell line Daudi, MM cell lines U266, KMS-26, ARP-1, KMS-11, KMS-12-BM, and MM.1S, PDAC cell lines CFPAC-1, PANC-1, MIA PaCa-2, and PL45, NSCLC cell lines NCI-H441 and NCI-H266, and TNBC cell lines MDA-MB-231 and MDA-MB-468 were purchased from American Type Culture Collection or obtained from Arkansas Cancer Research Center.

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test, Control

Journal: Nature Communications

Article Title: T cells engineered against Dickkopf-1-A2 complex can be used to treat HLA-A2 + solid and hematologic cancers

doi: 10.1038/s41467-026-69621-8

Figure Lengend Snippet: a Expression of DKK1, HLA-A2, and DKK1-A2 complex by primary tumor cells of 7 randomly selected PDAC patients determined by flow cytometry. b CAR-T cell proliferation measured by dilution of Celltrace Violet, analyzed by flow cytometry. CAR-T cells were stained with Celltrace Violet and co-cultured with primary PDAC tumor cells from the patients at an E:T ratio of 1:1. c Representative flow staining, and d Summarized results depicting the in vitro cytolytic activity of CAR-T cells against primary PDAC cells determined by flow cytometry, measuring the number or percentage of primary tumor cells and CAR-T cells in their co-culture. Primary PDAC tumor cells were stained with Celltrace Violet and co-cultured with CAR-T cells for 72 h. n = 2, technical replicates. One-way ANOVA was used. e – g Percentages of IFN-γ, TNF-α, and Granzyme B producing cells determined by intracellular cytokine staining. CAR-T cells were co-cultured with primary PDAC tumor cells at an E:T ratio of 1:1 overnight. n = 4, biological replicates. Two-tailed Student’s t -test was used. h Schema of PDAC PDX model treated with CAR-T cells, i Tumor volume of PDAC PDX mice measured weekly. Two-way ANOVA was used, j Survival of PDAC PDX mice monitored daily, Log-rank test was used, and k Percentages of CAR-T cells in blood, spleen, and bone marrow in PDAC PDX mice identified as GFP expression by flow cytometry before and post-CAR-T cell infusion. Percentage (%) was defined as the proportion of eGFP + CAR-T cells in blood cells, splenocytes, or bone marrow cells. Ctrl, n = 5; CD19 CAR-T, n = 5; mC2 CAR-T, n = 5; biological replicates. Two-tailed Student’s t -test was used. In some experiments, CD19 CAR-T cells were used as a control. Data shown as mean ± SEM. Source data are provided as a file.

Article Snippet: Human mantle cell lymphoma cell line JeKo-1, Burkitt’s lymphoma cell line Daudi, MM cell lines U266, KMS-26, ARP-1, KMS-11, KMS-12-BM, and MM.1S, PDAC cell lines CFPAC-1, PANC-1, MIA PaCa-2, and PL45, NSCLC cell lines NCI-H441 and NCI-H266, and TNBC cell lines MDA-MB-231 and MDA-MB-468 were purchased from American Type Culture Collection or obtained from Arkansas Cancer Research Center.

Techniques: Expressing, Flow Cytometry, Staining, Cell Culture, In Vitro, Activity Assay, Co-Culture Assay, Two Tailed Test, Control

Journal: Nature Communications

Article Title: T cells engineered against Dickkopf-1-A2 complex can be used to treat HLA-A2 + solid and hematologic cancers

doi: 10.1038/s41467-026-69621-8

Figure Lengend Snippet: a , b In vitro cytotoxicity of CAR-T cells determined by luciferase reporter assay. CAR-T cells were co-cultured with U266- or CFPAC-1-Luc tumor cells at indicated E:T ratios for 12 h. n = 2, technical replicates. c Representative flow staining, and d summarized results depicting the in vitro cytolytic activity of CAR-T cells against primary PDAC cells determined by flow cytometry, measuring the number or percentage of primary tumor cells and CAR-T cells in their co-culture. Primary PDAC tumor cells were stained with Celltrace Violet and co-cultured with CAR-T cells for 24 or 72 h. n = 3, technical replicates. One-way ANOVA was used. e Schema of U266 MM xenograft model treated with CAR-T cells, f Tumor burden measured as the level of human Igλ secreted by U266 cells. Blood was collected from U266 xenograft mice weekly, and serum was used for ELISA, Two-way ANOVA was used, and g Survival of U266 MM xenograft mice monitored daily before and post CAR-T cell infusion. Log-rank test was used. Ctrl, n = 4; mC2 CAR-T, n = 5; hC2 CAR-T, n = 5; biological replicates. h Schema of CFPAC-1-Luc PDAC orthotopic xenograft model treated with CAR-T cells, i Tumor burden measured as bioluminescence intensity of CFPAC-1-Luc PDAC orthotopic xenograft mice, Two-way ANOVA was used, j Survival of CFPAC-1-Luc PDAC orthotopic xenograft mice monitored daily, Log-rank test was used, and k Bioluminescence images of CFPAC-1-Luc PDAC orthotopic xenograft mice before and post CAR-T cell infusion. Ctrl, n = 4; mC2 CAR-T, n = 4; hC2 CAR-T, n = 4; biological replicates. Data shown as mean ± SEM. Source data are provided as a file.

Article Snippet: Human mantle cell lymphoma cell line JeKo-1, Burkitt’s lymphoma cell line Daudi, MM cell lines U266, KMS-26, ARP-1, KMS-11, KMS-12-BM, and MM.1S, PDAC cell lines CFPAC-1, PANC-1, MIA PaCa-2, and PL45, NSCLC cell lines NCI-H441 and NCI-H266, and TNBC cell lines MDA-MB-231 and MDA-MB-468 were purchased from American Type Culture Collection or obtained from Arkansas Cancer Research Center.

Techniques: In Vitro, Luciferase, Reporter Assay, Cell Culture, Staining, Activity Assay, Flow Cytometry, Co-Culture Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: AUM-302 exerts a more substantial inhibitory growth effect in PDAC spheroids than other tested compounds. (A) hF37 PDAC organoid-derived 2D cell line, (B) BxPC-3, (C) Capan-2, (D) MIA PaCa-2, and (E) PANC-1 PDAC cell lines were treated as spheroids with DMSO (vehicle) and 1 μM and 10 μM concentrations of BEZ-235, GDC-0941, TP-3654, or AUM-302 for 72 h. The figure displays representative images from a sample of N = 5 . The scale bar depicts 100 μm.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques: Derivative Assay

Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: BEZ-235, GDC-0941, TP-3654, and AUM-302 inhibit the viability of multiple pancreatic cancer cell lines in a spheroid setting. Pancreatic cancer cell lines BxPC-3 (A) Capan-2 (B) MIA PaCa-2 (C) PANC-1 (D) and organoid-derived cell line hF37 2D (E) were treated with variable concentrations of BEZ-235 ( A small solid pink circle on a white background. ) GDC-0941( A plain blue square with a solid color. ) TP-3654( Black triangle with a solid fill. ) and AUM-302 ( A green downward-facing triangle on a white background. ) for 72 h, and cell viability was measured using 3D Cell Titer-Glo. The results are shown as mean ± SD ( N = 5 ) and normalized to the control (DMSO). The LOG -9 represents 1 nM, -8 – 10 nM, -7 – 100nM, -6 – 1 μM, -5 – 10 μM, -4 – 100 μM.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques: Derivative Assay, Control

Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: AUM-302 exerts a more substantial inhibitory growth effect in hF31, hF37, hF44, hT1, and hM1a PDAC organoids than other tested compounds. hF31 (A–D) hF37 (E–H) hF44 (I–L) hT1 (M–P) and hM1a (Q–T) PDAC organoids were seeded in Matrigel and treated with DMSO (vehicle) and 10 nM, 100 nM, and 1 μM concentrations of GDC-0941 (A,E,I,M,Q) TP-3654 (B,F,J,N,R) BEZ-235 (C,G,K,O,S) or AUM-302 (D,H,L,P,T) for 72 h. The figure displays representative images from a sample of N = 5 . The scale bar depicts 100 μm.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: AUM-302 significantly exerts a stronger inhibitory growth effect on PDAC organoids than BEZ-235. hF31 (A) hF37 (B) hF44 (C) hT1 (D) and hM1a (E) PDAC organoids were seeded in Matrigel and treated with DMSO (vehicle) and 10 nM, 100 nM, and 1 μM concentrations of BEZ-235 and AUM-302 for 72 h. At the end of the incubation, cell viability was measured using 3D Cell Titer-Glo. The results are shown as mean ± SD ( N = 5 ) and normalized to the control (DMSO). The statistical analysis was performed using a two-way ANOVA in GraphPad Prism version 10.4.1. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques: Incubation, Control

Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: AUM-302 synergizes with RMC-6236 to inhibit the growth of hT1 PDAC organoids. hT1 PDAC organoids were seeded in Matrigel and treated with various concentrations of AUM-302, RMC-6236, and a combination of both compounds. Seventy-two hours post-treatment, the cell viability was assessed using 3D Cell Titer-Glo and analyzed using a non-linear regression model of log(inhibitor) vs. response–variable slope (four parameters) equation in GraphPad Prism version 10.4.1. and Synergy Finder. (A) Cell viability of mono-and combinatorial treatment. The results are shown as mean ± SD ( N = 5 ) and normalized to the control (DMSO). (B) The HSA Synergy Score is a dose matrix inhibition response. (C) The HSA Synergy Score is shown as a plot contour. (D) The graph was generated using a Plot Barometer using viability results. The needle points to the observed growth inhibition, denoted in percentage values, for the synergy between AUM-302 0.1 μM and RMC-6236 0.33 μM. The four white lines represent the predicted inhibition values calculated by the HSA, Loewe, ZIP, and BLISS models at specific concentrations.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques: Control, Inhibition, Generated

Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: AUM-302 and RMC-6236 affected the activity of the signaling pathways in the hT1 PDAC organoids. hT1 organoids were treated with DMSO (vehicle), 300nM RMC-6236, 10 nM and 100 nM AUM-302, and combinations of thereof for 24 and 48 h. (A) Western blots were performed for mTOR, AKT, ERK, S6, phosphorylated counterparts, and β-Actin as a loading control. (B,C) Densitometry analysis was conducted to quantify relative protein expression, normalized to β-actin control and non-phosphorylated proteins in samples after 24- and 48-h post-treatment. Densitometry was measured with ImageJ and analyzed with GraphPad Prism as described in “Materials and Methods.” Data are presented as mean ± SD values ( N = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques: Activity Assay, Protein-Protein interactions, Western Blot, Control, Expressing

Journal: Nature Communications

Article Title: Combined targeted and epigenetic-based therapy enhances antitumor immunity by stabilizing GATA6-dependent MHCI expression in pancreatic ductal adenocarcinoma

doi: 10.1038/s41467-026-69013-y

Figure Lengend Snippet: a Nine primary murine PDAC cell lines were established from spontaneous murine PDAC models ( CKP , KPC , and KPCY ). Created with BioRender.com. Flow cytometry analysis was used to determine GATA6 protein expression in each line. Representative histograms show GATA6 staining (GATA6 antibody, GATA6 AB, red) versus isotype control (blue) in GATA6 high (GATA6 hi ) and low (GATA6 lo ) expressing cell lines. The cell lines were dichotomized into GATA6 high ( n = 5, orange) and GATA6 low ( n = 4, blue) groups. b The parental cells (Ctrl) of three GATA6 high cell lines (60400, 511950, 70301) and three GATA6 low cell lines (60590, 60531, 511892) were treated with increasing doses of MEKi (trametinib) until 100x of the cells’ initial IC50 to generate the corresponding long-term MEKi-treated cells (MEKi). Transcriptomic profiling by RNA sequencing of the parental ( Ctrl) and long-term MEKi-treated (MEKi) cells was performed. GSEA of KEGG and HALLMARKs revealed gene sets that were significantly ( P -adjust < 0.05) different between parental (Ctrl) and long-term MEKi-treated (MEKi) cells within GATA6 high and GATA6 low groups, respectively. c Surface MHCI (H-2Db) expression on control (Ctrl) and MEKi-treated (MEKi) cells of GATA6 high (upper panel) and GATA6 low group (lower panel) was assessed by flow cytometry ( n = 4 biological replicates for cell line 60400, 511950, 511892, 60590, and 5 biological replicates for cell line 110299, 2838c3, 60531, 6694C2). MFI mean fluorescence intensity. Mean ± SD is shown. Statistical significance was calculated by unpaired two-tailed Mann–Whitney test. d Flow cytometric analysis of GATA6 expression in GATA6 knockout (KPCY1-CRISPR-GFP v2.1-gRNA-gCas9): gGATA6_KO-3 and gGATA6_KO-4, and negative control (gNT) cell lines ( n = 4 biological replicates). Statistical significance was calculated by unpaired two-tailed Mann–Whitney test. e Surface MHCI (H-2Db) expression on GATA6 knockout cell lines and negative control upon MEKi treatment ( n = 4 biological replicates). MFI: mean fluorescence intensity. Mean ± SD is shown. Statistical significance was calculated by unpaired two-tailed Mann–Whitney test. f Schematic of the knock-in strategy for N-terminal AID-tagged gata6. Shown are components of the knock-in cassette, and the positions of primers for genomic PCR are marked by arrows. g Western blot showing GATA6 protein expression in 110299 WT and 110299 AID-GATA6 cells after 24 h treatment with 1 µM 5-Ph-IAA ( n = 1 biological replicates). h Surface MHCI (H-2Db) expression on 110299 WT and 110299 AID-GATA6 cells treated with or without MEKi and/or 1 µM 5-Ph-IAA ( n = 5 biological replicates). MFI: mean fluorescence intensity. Mean ± SD is shown. Statistical significance was calculated by One-way ANOVA and Kruskal–Wallis test.

Article Snippet: HAs were PCR-amplified using murine primary PDAC cell line (110299) genomic DNA as template and cloned into pJET (Thermo Fisher Scientific) flanking the Blast-P2A-V5-AID cassette to generate pJET_gata6_N-AID_HDR.

Techniques: Flow Cytometry, Expressing, Staining, Control, RNA Sequencing, Fluorescence, Two Tailed Test, MANN-WHITNEY, Knock-Out, CRISPR, Negative Control, Knock-In, Western Blot

Journal: Nature Communications

Article Title: Combined targeted and epigenetic-based therapy enhances antitumor immunity by stabilizing GATA6-dependent MHCI expression in pancreatic ductal adenocarcinoma

doi: 10.1038/s41467-026-69013-y

Figure Lengend Snippet: a Patient-derived xenografts (PDXs) from 15 PDAC patients were transplanted subcutaneously into athymic nude mice, and the mice of third generation were used for MEKi treatment. When tumor volume reached 200 mm 3 , the mice were treated with vehicle control (Ctrl) or MEKi. Tumors were harvested after 4 weeks of treatment. Created with BioRender.com. b IHC staining of GATA6 and MHCI of the xenograft tumors was performed, and the proportion of marker-positive cells out of total cells was quantified by Definiens software. Xenografts were dichotomized into GATA6 high (GATA6 hi , n = 7 independent mice) and GATA low (GATA lo, n = 8 independent mice) expression groups using median GATA6 + cells (% total) as cutoff. MHCI + cells (% total) were quantified by Definiens and compared between vehicle control and corresponding MEKi-treated mice in GATA6 high and GATA low expression groups, respectively. Statistical significance was calculated by two-tailed Wilcoxon matched-pairs signed rank test. c IHC staining showing reduction of GATA6 expression in MEKi-treated tumors in GATA6 high expression group. Signal intensities were quantified by Definiens and compared between vehicle control and MEKi-treated mice groups ( n = 7 independent mice for each group). Statistical significance was calculated by two-tailed Wilcoxon matched-pairs signed rank test. d Flow cytometry analysis of GATA6 expression in 110299 cells treated with MEKi, class I HDAC inhibitors (mocetinostat, quisinostat, domatinostat), class II HDAC inhibitors (tasquinimod, LMK235, ricolinostat), or their combinations ( n = 4 independent experiments for each cell line). Data are presented as mean values ± SD. Individual dots represent independent biological replicates. Statistical significance was calculated by One-way ANOVA, Kruskal–Wallis test. e Western blot analysis of GATA6 protein levels in four GATA6 high (orange: 110299, 511950, 2838c3, 60400) and four GATA6 low (blue: 60531, 511892, 60590, 6694c2) murine PDAC cell lines following treatment with MEKi and/or domatinostat; β-actin served as a loading control ( n = 1 independent experiment). f Surface MHCI (H-2Db) expression in the four GATA6 high (orange) and four GATA6 low (blue) murine PDAC cell lines treated with MEKi and/or domatinostat, assessed by flow cytometry ( n = 4 independent experiments for cell line 511950, 60400, 511892, 60590, and 5 independent experiments for cell line 110299, 2838c3, 60531, 6694C2). Data are presented as box plots: the center line indicates the median, the bounds of the box indicate the 25th and 75th percentiles, and whiskers extend to minima and maxima. One-way ANOVA and Kruskal–Wallis test were used. Ctrl: vehicle control; MEKi: trametinib; Moce: mocetinostat; Quis: quisinostat; Doma: domatinostat; Tas: tasquinimod; LMK: LMK235; Rico: ricolinostat. MFI mean fluorescence intensity. Scale bar: μm.

Article Snippet: HAs were PCR-amplified using murine primary PDAC cell line (110299) genomic DNA as template and cloned into pJET (Thermo Fisher Scientific) flanking the Blast-P2A-V5-AID cassette to generate pJET_gata6_N-AID_HDR.

Techniques: Derivative Assay, Control, Immunohistochemistry, Marker, Software, Expressing, Two Tailed Test, Flow Cytometry, Western Blot, Fluorescence