高分文献深度解读｜攻坚生命科学前沿研究，解析核心分子作用机制

Fig. 1. STING pathway activation enhances the antitumor efficacy of YM101.

A Tumor growth curves of EMT-6-bearing mice treated with control IgG (CTL) or YM101. Tumors were collected on day 19 (n?=?8 for CTL; n?=?9 for YM101). B Tumor growth inhibition (TGI) of individual EMT-6 tumors; TGI?<?50% was defined as nonresponder, and TGI?≥?50% as responder. C GSEA of the GO term “innate immune response” comparing YM101 responders versus nonresponders in EMT-6 tumors. D Top enriched immune/cytokine GO terms ranked by normalized enrichment score (NES); all terms shown have FDR?<?0.05. E Tumor growth curves of CT26-bearing mice treated with CTL or YM101. Tumors were collected on day 18 (n?=?6 for CTL; n?=?15 for YM101). F TGI of individual CT26 tumors. G, H GSEA of the GO terms “innate immune response” and “response to IFN-β” comparing YM101 responders versus nonresponders in CT26 tumors. I Overlap of upregulated DEGs (UpDEGs) in YM101-responsive EMT-6 and CT26 tumors; representative genes are listed. J Functional enrichment analysis of overlapping upregulated DEGs. K Intratumoral cytokine profile in EMT-6 tumors from an independent cohort after YM101 treatment (n?=?6 for responders; n?=?5 for nonresponders). L, O Representative tumor images from EMT-6 and CT26 models treated as indicated. M, P Tumor growth curves of EMT-6 and CT26 models (n?=?8 mice per group). N, Q Kaplan–Meier survival curves for EMT-6 and CT26 models (n?=?8 mice per group). A, E Data are mean?±?SD. The P values shown were calculated using two-tailed unpaired t-tests. M, P Data are mean?±?SD. Statistical comparisons were performed using one-way ANOVA followed by Holm–?ídák’s multiple comparisons test. N, Q P values were determined by the log-rank test. Statistical significance: *P?<?0.05; **P?<?0.01; ***P?<?0.001; ****P?<?0.0001. Source data and exact P values are provided in the Source data file.

Fig. 3. Combination therapy (MSA-2?+?YM101) enhances CXCL16–CXCR6 signaling in the tumor microenvironment.

For scRNA-seq analyses, n?=?3 independent pooled samples per group (each sample pooled from tumors of two biologically independent mice; 6 mice per group in total). A Scatter plots comparing the outgoing (x-axis) and incoming (y-axis) interaction strengths of T cells in combo vs. CTL, combo vs. MSA-2, and combo vs. YM101. The CXCL signaling pathway is strengthened in the combo group. B, C Chord diagrams and circular plots illustrating CXCL signaling networks among various cell types in CTL-, MSA-2-, YM101-, and combo-treated tumors. Thicker chords and circles indicate stronger signaling contributions. D Heatmaps showing the communication probability of CXCL signals originating from macrophages and cDCs under each treatment. E Bubble plots identifying potential ligand-receptor pairs contributing to the enhanced CXCL signaling in Combo-treated tumors. Dot color represents communication probabilities, and dot size reflects statistical significance (dots are absent when no communication is inferred). F, G Chord diagrams and circular plots specifically highlighting the CXCL16–CXCR6 axis. H Network analysis mapping the source and target cells involved in CXCL16–CXCR6 signaling. I Venn diagram displaying the number of significantly upregulated genes (UpDEGs) in macrophages from Combo-treated tumors compared to CTL, MSA-2, and YM101 treatments. J Heatmap showing representative immune-related genes with increased expression in macrophages under Combo therapy. K Box plots showing Cxcl16 expression across indicated macrophage subsets in EMT-6 tumors from scRNA-seq data under CTL, MSA-2, YM101, or combination treatment. Data are shown for all macrophages from three pooled samples per group (n?=?3). Single cells are shown for visualization only and are not considered independent biological replicates. Box plots show the median (center line), 25th–75th percentiles (box), and whiskers extending to the most extreme values within 1.5?×?IQR; outliers are shown as individual points.

Fig. 4. Triple-targeting therapy increases tumor-infiltrating CXCR6+CD8+ T cells and CXCL16+ myeloid cells in the EMT-6 model.

A Flow cytometry (FC) analysis of the relative abundance and fold changes of tumor-infiltrating CD8? T cell subsets across different treatment conditions. Tumors were obtained from an independent cohort in which treatment was initiated when tumors reached 200–250?mm3, and tumors were collected 24?h after completion of the dosing schedule for subsequent flow cytometry analyses (n?=?4 per group). B–E Percentage plots showing increased expression of Granzyme-B, Perforin, CD69, and Ki67 in CXCR6? T cells compared to CXCR6? T cells (n?=?32 paired samples). F Gating strategy for isolating CXCR6?CD8? T cells from tumor-infiltrating lymphocytes for downstream proteomic analyses. G Heatmap of differentially expressed proteins comparing CXCR6? and CXCR6?CD8? T cells (n?=?2 per group). H Enrichment analysis of differentially expressed proteins in CXCR6? versus CXCR6?CD8? T cells. I Percentage plots demonstrating higher PD-1 expression in CXCR6?CD8? T cells relative to CXCR6?CD8? T cells (n?=?32 paired samples). J Co-culture experiment using CXCR6? or CXCR6?CD8? T cells (from OT-1 mice) with EG7-OVA cells, showing enhanced tumor cell death induced by CXCR6?CD8? T cells compared to CXCR6? T cells. Anti-PD-L1 treatment further increases cytotoxic activity (n?=?3 biological replicates; results are representative of three independent experiments). K Flow cytometry analysis of myeloid cell populations in the EMT-6 model, showing the scaled number of CXCL16+ macrophages (CXCL16+ M?) and dendritic cells (CXCL16+ DC) following treatment with the specified therapies (n?=?4 per group). Statistical comparisons were made between the M?+?Y group and all other treatment groups unless otherwise specified. B–E, I Were analyzed using two-tailed paired t-tests. A, J, K Were analyzed using one-way ANOVA followed by Holm–?ídák’s multiple comparisons test. Statistical significance is indicated as *P?<?0.05; **P?<?0.01; ***P?<?0.001; ****P?<?0.0001. Source data and exact P values are provided in the Source data file.