ANGPTL3 in the peripheral circulation is associated with resistance to anti-PD1 therapy in advanced gastric cancer

Figures

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  Figure 1.. ANGPTL3 levels are high in the peripheral blood of patients with AGC who progress after nivolumab treatment. Peripheral blood was collected from patients with AGC before and 1 month after initiating nivolumab therapy, and ANGPTL3 levels in plasma were measured by proteomic profiling or ELISA (n = 80). A, ANGPTL3 seq.10382-1 detected by proteomic profiling. B, ANGPTL3 seq.10391-1 detected by proteomic profiling. C, ANGPTL3 protein detected by ELISA. The median values for each group are shown in brackets. No significant differences were observed between pretreatment and posttreatment levels for any of the markers.

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  Figure 2.. High plasma ANGPTL3 levels are significantly associated with poor prognosis of patients with AGC after nivolumab treatment. Patients were divided into two groups, high (red lines) and low (blue lines), based on ANGPTL3 cutoff values before treatment, and significant differences in PFS and OS were statistically analyzed by the Kaplan–Meier method and the Mantel–Cox log-rank test. A, ANGPTL3 seq.10382-1 detected by proteomic profiling. B, ANGPTL3 seq.10391-1 detected by proteomic profiling. C, ANGPTL3 protein detected by ELISA.

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  Figure 3.. High posttreatment ANGPTL3 levels are significantly associated with shorter OS of patients with AGC after nivolumab treatment. Patients were divided into two groups, high (red lines) and low (blue lines), based on ANGPTL3 cutoff values after treatment, and significant differences in PFS and OS were statistically analyzed by the Kaplan–Meier method and the Mantel–Cox log-rank test. A, ANGPTL3 seq.10382-1 detected by proteomic profiling. B, ANGPTL3 seq.10391-1 detected by proteomic profiling. C, ANGPTL3 protein detected by ELISA.

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  Figure 4.. Blocking ANGPTL3 synergistically enhances anti-PD1 therapeutic efficacy in mouse tumor models. BALB/c mice were subcutaneously (5 × 105) implanted with Colon26 cells, and received intraperitoneal injection of anti-ANGPTL3 mAb, anti-PD1 mAb, or mIgG as a control at 10 mg/kg on days 3 and 10 after tumor implantation (n = 5). On day 17, the subcutaneous tumor and spleen were harvested randomly from three of 10 mice for assays. A, Suppression of tumor growth by treatments (n = 10). Open circles, mIgG control; closed circles, anti-ANGPTL3 mAb; and closed triangles, anti-PD1 mAb. Gray diamonds, anti-ANGPTL3 mAb and anti-PD1 mAb. B, Dramatical increase of tumor-infiltrating lymphocytes (TIL) by anti-ANGPTL3/-PD1 combination therapy. Tumor-infiltrating lymphocytes were analyzed by flow cytometry (n = 3). C, Increase of SPCs by treatments. SPCs were analyzed by flow cytometry (n = 3). D, Enhancement of the cytotoxic activity of splenic CD8+ T cells by anti-ANGPTL3/-PD1 combination therapy. AH1-prestimulated CD8+ T cells were cocultured with Colon26 cells as a target at effector:target (E:T) ratio = 40:1 for 4 hours (n = 3). Splenic DX5+ NK cells were also cocultured with Yac1 cells as a…