Fig. 6: GDF1-induced tumour-lineage plasticity might sensitise HCC patients to anti-PD1 therapy.

a HCC patients from the TCGA–LIHC project were divided into three subgroups according to the specified classifiers. The relative expression of GDF1, CD8A, TGFB1, and PDL1 was shown in different subgroups of patients (one-way ANOVA test). b C57BL/6 mice bearing Hepa1–6 tumours transfected with mGDF1 or control vector were treated with anti-PD1 antibodies or control IgG at 100 μg/mouse every three days. The tumour-inhibition rates were detected by an in vivo bioluminescence imaging system (two-way ANOVA test, data are presented as mean values ± SEM, CTR-IgG: n = 6, CTR-PD1: n = 6, GDF1-IgG: n = 8, GDF1-PD1: n = 8). c Kaplan–Meier analysis of overall survival in C57BL/6 mice bearing Hepa1-6 tumours transfected with mGDF1 or control vector and treated with anti-PD1 antibodies or control IgG (log-rank test). d IHC staining of CD8 and GZMB in mouse orthotopic Hepa1–6 tumours transfected with mGDF1 or control vector and treated with anti-PD1 antibodies or control IgG. Scale bars represent 50 µm, n = 3 independent experiments. e, f IHC staining of GDF1 and CD8 + infiltrating T lymphocytes was performed in consecutive TMA slides. Kaplan–Meier analysis of overall survival (e) and disease-free survival (f) in HCC patients divided into four subgroups according to GDF1 and CD8 staining (NS, not significant, log-rank test). g Representative IHC staining of GDF1 and CD8 in consecutive TMA slides with 196 liver tumour tissues. Scale bars represent 50 µm. IgG, immunoglobulin G. PD1, anti-PD1 antibody. h C57BL/6 mice bearing Hepa1–6-CTR or Hepa1–6-GDF1^KO cells were treated with LSD1 inhibitor, anti-PD1 antibody, or combination of both. For treatment with antibody, tumour-bearing mice were treated with anti-PD1 antibodies or control IgG at 100 μg/mouse every three days. For LSD1 inhibitor treatment, tumour-bearing mice were treated with 10 mg/kg of GSK-LSD1 or vehicle (4% DMSO in saline) each week (4 consecutive days followed by a 3-day holiday). The tumour inhibition rates were detected by an in vivo bioluminescence imaging system (NS, not significant, two-way ANOVA test, data are presented as mean values ± SEM, n = 8 mice per group). i Kaplan–Meier analysis of overall survival in C57BL/6 mice bearing Hepa1–6 tumours with different treatment groups (log-rank test). j The number of metastatic lung nodules in mice bearing Hepa1–6 tumours with different treatment groups. (NS, not significant, two-tailed independent Student’s t-test, data are presented as mean values ± SEM, n = 8 mice per group). Source data are provided as a Source Data file.