Journal: Neoplasia (New York, N.Y.)

Article Title: OSMR coordinates a self-perpetuating circuit linking chemoresistance and neutrophil-driven immunosuppression in gastric cancer

doi: 10.1016/j.neo.2026.101279

Figure Lengend Snippet: OSMR increases BMP5 expression by inhibiting TRIM2 which regulates GATA3 ubiquitination. (A) Genes enriched in the Cytokine-cytokine receptor interaction signaling pathway. (B) BMP5 mRNA and protein expression in AGS and MKN28 cells when OSMR was up-regulated or down-regulated. (C) Correlation between BMP5 expression and GC patients’ overall survival in Kaplan-Meier plotter database. (D) Jaspar and hTFtarget database prediction of the transcription factors of BMP5. (E) BMP5 mRNA expression when four transcription factors were down-regulated. (F) Dual-Luciferase Reporter Assay analysis for GATA3-binding BMP5 promoter. (G) GATA3 mRNA and protein expression in AGS and MKN28 cells when OSMR was up-regulated or down-regulated. (H) GATA3 protein expression in MKN28 cells when CHX was treated. (I) GATA3 Ubiquitination detection when OSMR was down-regulated. (J) TRIM family expression in AGS when OSMR was overexpressed. (K) TRIM2, GATA3, and BMP5 expression in AGS cells (left); Co-IP analysis of interaction between OSMR and PI3K (right). (L) GATA3 Ubiquitination detection when TRIM2 was down-regulated. (**, P < 0.01).

Article Snippet: The membranes were blocked with 5% non-fat milk in TBST for 1 hour at room temperature, followed by an overnight incubation at 4°C with primary antibodies (OSMR (Abcam, #ab315388), CCNE2 (Abcam, #ab40890), β-actin (CST, #4967), PI3K (P85) (CST, #4292), p-PI3K (p-P85) (Abcam, #ab278545), AKT (CST, #9272), p-AKT (CST, #4060), P27 (CST, #2552), p-P27 (Abcam, #ab62364), FLAG (CST, #14793), BMP5 (Proteintech, #13253-1-AP), Ub (CST, #20326), TRIM2 (Proteintech, #20356-1-AP), GATA3 (CST, #5852), p-SMAD1/5 (CST, #9516), SMAD5 (CST, #12534), PD-L1 (CST, #13684), BATF (CST, #8638), IL-31 (abcam, #ab62579)), which were diluted in accordance with the manufacturer's instructions.

Techniques: Expressing, Ubiquitin Proteomics, Luciferase, Reporter Assay, Binding Assay, Co-Immunoprecipitation Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: OSMR coordinates a self-perpetuating circuit linking chemoresistance and neutrophil-driven immunosuppression in gastric cancer

doi: 10.1016/j.neo.2026.101279

Figure Lengend Snippet: OSMR induced BMP5 promotes N2 polarization of neutrophils and inhibits CD8+ T cells function. (A) Photos, weights and volume of tumors from MFC cells knocking down BMP5. The animal model used was 615 mice. (B) Ki67 staining of tumors from MFC cells knocking down BMP5. (C) Gating strategy for the flow cytometry experiments. (D) Percentage of immune cells in CD45 + cells in tumors from MFC cells knocking down BMP5. (E, F) The expression of N1 and N2 biomarkers in neutrophils after co-cultured with MKN28 cells.The Neutrophils used are derived from humans. (G, H) The expression of N1 and N2 biomarkers in neutrophils after co-cultured with MKN28 cells and treated with BMP5. (I, J) TNF-α and IFN-γ expression in CD8+ T cells after co-cultured neutrophils and GC cells. (*, P < 0.05; **, P < 0.01).

Article Snippet: The membranes were blocked with 5% non-fat milk in TBST for 1 hour at room temperature, followed by an overnight incubation at 4°C with primary antibodies (OSMR (Abcam, #ab315388), CCNE2 (Abcam, #ab40890), β-actin (CST, #4967), PI3K (P85) (CST, #4292), p-PI3K (p-P85) (Abcam, #ab278545), AKT (CST, #9272), p-AKT (CST, #4060), P27 (CST, #2552), p-P27 (Abcam, #ab62364), FLAG (CST, #14793), BMP5 (Proteintech, #13253-1-AP), Ub (CST, #20326), TRIM2 (Proteintech, #20356-1-AP), GATA3 (CST, #5852), p-SMAD1/5 (CST, #9516), SMAD5 (CST, #12534), PD-L1 (CST, #13684), BATF (CST, #8638), IL-31 (abcam, #ab62579)), which were diluted in accordance with the manufacturer's instructions.

Techniques: Animal Model, Staining, Flow Cytometry, Expressing, Cell Culture, Derivative Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: OSMR coordinates a self-perpetuating circuit linking chemoresistance and neutrophil-driven immunosuppression in gastric cancer

doi: 10.1016/j.neo.2026.101279

Figure Lengend Snippet: Tumor BMP5 activates the SMAD1/5 pathway in neutrophils and promotes PD-L1 transcription. (A) Correlation between neutrophil score and BMP5 receptors in TCGA database. (B) Schematic of BMP5 and its receptors signaling pathway. (C) p-SMAD1/5 and SMAD protein levels in neutrophils. (D) PD-L1 mRNA and protein level in neutrophils treated with BMP5 and SMAD1/5 inhibitor. (E) PD-L1 mRNA and protein level in neutrophils co-cultured with MKN28 cells or treated with SMAD1/5 inhibitor. (F) Dual-Luciferase Reporter Assay analysis for SMAD5-binding PD-L1 promoter. (G) mIF staining and correlation of BMP5,CD15, PD-L1, and CD8 in tumor samples from patients in Cohort 2. (**, P < 0.01).

Article Snippet: The membranes were blocked with 5% non-fat milk in TBST for 1 hour at room temperature, followed by an overnight incubation at 4°C with primary antibodies (OSMR (Abcam, #ab315388), CCNE2 (Abcam, #ab40890), β-actin (CST, #4967), PI3K (P85) (CST, #4292), p-PI3K (p-P85) (Abcam, #ab278545), AKT (CST, #9272), p-AKT (CST, #4060), P27 (CST, #2552), p-P27 (Abcam, #ab62364), FLAG (CST, #14793), BMP5 (Proteintech, #13253-1-AP), Ub (CST, #20326), TRIM2 (Proteintech, #20356-1-AP), GATA3 (CST, #5852), p-SMAD1/5 (CST, #9516), SMAD5 (CST, #12534), PD-L1 (CST, #13684), BATF (CST, #8638), IL-31 (abcam, #ab62579)), which were diluted in accordance with the manufacturer's instructions.

Techniques: Cell Culture, Luciferase, Reporter Assay, Binding Assay, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: OSMR coordinates a self-perpetuating circuit linking chemoresistance and neutrophil-driven immunosuppression in gastric cancer

doi: 10.1016/j.neo.2026.101279

Figure Lengend Snippet: Tumor induced PD-L1 expression in neutrophils enhances IL31 secretion of CD8 + T cells. (A) IL31 and OSM mRNA level in CD8 + T cells after co-cultured neutrophils and GC cells. (B) TIM-3, PD-1 and IL31 mRNA level in CD8 + T cells treated with IL2 and Anti-CD3/CD28 antibody. (C) IL31 mRNA and protein level in CD8 + T cells co-cultured with neutrophils treated with BMP5 and PD-L1 inhibitor (Atezolizumab). (D) Schematic of PD-1/PD-L1 signaling pathway in CD8 + T cells. (E) BATF mRNA and protein level in CD8 + T cells treated with IL2 and Anti-CD3/CD28 antibody for 0 day and 14 days. (F) BATF mRNA and protein level in CD8 + T cells after co-cultured neutrophils and GC cells. (G) IL31 mRNA and protein level in CD8 + T cells when BATF was up-regulated or down-regulated. (H) Dual-Luciferase Reporter Assay analysis for BATF-binding IL31 promoter. (I) p-PI3K, PI3K, p-AKT, AKT, p-P27, P27, CCNE2, PD-L1, and PD-L2 expression in MKN28 cells treated with supernatant of CD8 + T cells (J) mIF staining analysis of correlation of OSMR, CD15, CD8, and IL31 in tumor samples from patients in Cohort 2. (**, P < 0.01).

Article Snippet: The membranes were blocked with 5% non-fat milk in TBST for 1 hour at room temperature, followed by an overnight incubation at 4°C with primary antibodies (OSMR (Abcam, #ab315388), CCNE2 (Abcam, #ab40890), β-actin (CST, #4967), PI3K (P85) (CST, #4292), p-PI3K (p-P85) (Abcam, #ab278545), AKT (CST, #9272), p-AKT (CST, #4060), P27 (CST, #2552), p-P27 (Abcam, #ab62364), FLAG (CST, #14793), BMP5 (Proteintech, #13253-1-AP), Ub (CST, #20326), TRIM2 (Proteintech, #20356-1-AP), GATA3 (CST, #5852), p-SMAD1/5 (CST, #9516), SMAD5 (CST, #12534), PD-L1 (CST, #13684), BATF (CST, #8638), IL-31 (abcam, #ab62579)), which were diluted in accordance with the manufacturer's instructions.

Techniques: Expressing, Cell Culture, Luciferase, Reporter Assay, Binding Assay, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: OSMR coordinates a self-perpetuating circuit linking chemoresistance and neutrophil-driven immunosuppression in gastric cancer

doi: 10.1016/j.neo.2026.101279

Figure Lengend Snippet: Vixarelimab blocks OSMR to improve chemotherapy effectiveness and T cell function. (A) p-PI3K, PI3K, CCNE2, and BMP5 expression in MKN28 cells treated with Vixarelimab. (B) Cell proliferation and migration of MKN28 cells with Fluorouracil or Vixarelimab treatment. (C) TNF-α and IFN-γ expression in CD8 + T cells co-cultured with MKN28 cells with Fluorouracil or Vixarelimab treatment. The neutrophils and CD8 + T cells used are derived from humans. (D) Photos and weights of tumors from MKN28 cells with Fluorouracil or Vixarelimab treatment. (E) Volume of tumors from MKN28 cells with Fluorouracil or Vixarelimab treatment. The animal model used was nude mice. (F) Ki67 and Cleaved Casoase-3 staining of tumors from MKN28 cells with Fluorouracil or Vixarelimab treatment. (*, P < 0.05, **, P < 0.01).

Article Snippet: The membranes were blocked with 5% non-fat milk in TBST for 1 hour at room temperature, followed by an overnight incubation at 4°C with primary antibodies (OSMR (Abcam, #ab315388), CCNE2 (Abcam, #ab40890), β-actin (CST, #4967), PI3K (P85) (CST, #4292), p-PI3K (p-P85) (Abcam, #ab278545), AKT (CST, #9272), p-AKT (CST, #4060), P27 (CST, #2552), p-P27 (Abcam, #ab62364), FLAG (CST, #14793), BMP5 (Proteintech, #13253-1-AP), Ub (CST, #20326), TRIM2 (Proteintech, #20356-1-AP), GATA3 (CST, #5852), p-SMAD1/5 (CST, #9516), SMAD5 (CST, #12534), PD-L1 (CST, #13684), BATF (CST, #8638), IL-31 (abcam, #ab62579)), which were diluted in accordance with the manufacturer's instructions.

Techniques: Cell Function Assay, Expressing, Migration, Cell Culture, Derivative Assay, Animal Model, Staining