Journal: Advanced Science

Article Title: IL4I1⁺ Macrophages and TDO2⁺ Myofibroblasts Drive AhR‐Mediated Immunosuppression and Ferroptosis Resistance in Solid Predominant Lung Adenocarcinoma

doi: 10.1002/advs.202513606

Figure Lengend Snippet: IL4I1 + TAMs and TDO2 + myCAFs synergistically promote CD8 + T cell exhaustion and immunotherapy resistance. ( A) Flow cytometry analysis of CD8, GZMB, PD‐1, and TIM3 expression in murine CD8 + T cells co‐cultured with mCAFs and iBMDMs ( n = 3 per group). P ‐values were calculated with one‐way ANOVA test with Tukey's post‐hoc test. ( B) In vivo bioluminescence imaging showing the growth of orthotopic lung tumors constructed by LLC, mCAFs‐vector/TDO2, and iBMDMs on day 21 ( n = 5 per group). ( C) Normalized bioluminescence imaging quantification of orthotopic lung tumors ( n = 5 per group). P ‐values were calculated with Student's t ‐test. ( D) Representative multiplexed IHC staining of orthotopic lung tumors stained for Pan‐CK, IL4I1, TDO2, F4/80, and COL1A1. Scale bar = 10 µm. ( E) Representative IHC staining of orthotopic lung tumors stained for Ki‐67, CD163, CD206, CD86, CD8, PD‐1, and GZMB. Scale bar = 50 µm. ( F) Schematic outline showing anti‐PD‐1 treatment for subcutaneous xenograft tumor formed by mouse KP lung cancer cells, mCAFs‐vector/TDO2, and iBMDMs in C57BL/6 mice. ( G) Gross appearance of subcutaneous xenograft tumor ( n = 5 per group). ( H) Tumor weight of subcutaneous xenograft tumor formed by mouse KP lung cancer cells, mCAFs‐vector/TDO2, and iBMDMs in the treatment of IgG2α and anti‐PD‐1 ( n = 5 per group). P ‐values were calculated with one‐way ANOVA test with Tukey's post‐hoc test. ( I) Bar plot showing the frequency of CD8 + GZMB + T cells, CD8 + PD‐1 + T cells among CD8A + T cell, and CD8 + TIM3 + T cells ( n = 5 per group). P ‐values were calculated with one‐way ANOVA test with Tukey's post‐hoc test. ( J) The concentrations of IL4I1 in the LUAD patients accepted anti‐PD‐1/PD‐L1 treatment. Violin plot showing the concentrations of blood plasma IL4I1 in PR ( n = 8) and SD/PD ( n = 11) LUAD patients (left). P ‐values were calculated with Student's t ‐test. Receiver operating characteristic (ROC) curves showing the diagnostic accuracy of IL4I1 (right) for prediction of immunology response.

Article Snippet: On day 9 post‐implantation, mice were randomly divided into four groups ( n = 5 per group) and intraperitoneally administered either IgG2α isotype control (BE0089, RRID: AB_1107769, BioXcell) or anti‐PD‐1 antibody (200 μg, BE0146, RRID: AB_10949053, BioXcell) every three days, corresponding to the following treatment groups: (1) mCAF‐vector + iBMDMs + IgG2α, (2) mCAF‐vector + iBMDMs + anti‐PD‐1, (3) mCAF‐TDO2 + iBMDMs + IgG2α, and (4) mCAF‐TDO2 + iBMDMs + anti‐PD‐1.

Techniques: Flow Cytometry, Expressing, Cell Culture, In Vivo, Imaging, Construct, Plasmid Preparation, Immunohistochemistry, Staining, Clinical Proteomics, Diagnostic Assay

Journal: Advanced Science

Article Title: IL4I1⁺ Macrophages and TDO2⁺ Myofibroblasts Drive AhR‐Mediated Immunosuppression and Ferroptosis Resistance in Solid Predominant Lung Adenocarcinoma

doi: 10.1002/advs.202513606

Figure Lengend Snippet: Synergistic efficacy of AhR antagonist, ferroptosis inducer, and immunotherapy in LUAD treatment. (A) Schematic outline showing combination treatment regimen (CH‐223191, IKE, and anti‐PD‐1) for subcutaneous tumors constructed by mouse KP lung cancer cells, mCAFs‐TDO2, and iBMDMs. ( B–D) Gross appearance of the tumor mass ( B ), tumor weight ( C ), and kinetics of the tumor volume (mm 3 ) ( D ) were measured and documented for C57BL/6 mice in group ( n = 6 per group). ( E–G) Representing H&E staining ( E ), TUNEL staining ( F ), and 4‐HNE staining ( G ) of tumor tissues under different treatment regimens. Scar bar = 200 µm. ( H) Bar plot showing percentages of CD8 + GZMB + T cells, CD8 + PD‐1 + T cells, and CD8 + TIM3 + T cells under different treatment regimens ( n = 6 per group). P ‐values were calculated with one‐way ANOVA test with Tukey's post‐hoc test.

Article Snippet: On day 9 post‐implantation, mice were randomly divided into four groups ( n = 5 per group) and intraperitoneally administered either IgG2α isotype control (BE0089, RRID: AB_1107769, BioXcell) or anti‐PD‐1 antibody (200 μg, BE0146, RRID: AB_10949053, BioXcell) every three days, corresponding to the following treatment groups: (1) mCAF‐vector + iBMDMs + IgG2α, (2) mCAF‐vector + iBMDMs + anti‐PD‐1, (3) mCAF‐TDO2 + iBMDMs + IgG2α, and (4) mCAF‐TDO2 + iBMDMs + anti‐PD‐1.

Techniques: Construct, Staining, TUNEL Assay

Journal: Advanced Science

Article Title: IL4I1⁺ Macrophages and TDO2⁺ Myofibroblasts Drive AhR‐Mediated Immunosuppression and Ferroptosis Resistance in Solid Predominant Lung Adenocarcinoma

doi: 10.1002/advs.202513606

Figure Lengend Snippet: IL4I1 + TAMs and TDO2 + myCAFs synergistically promote CD8 + T cell exhaustion and immunotherapy resistance. ( A) Flow cytometry analysis of CD8, GZMB, PD‐1, and TIM3 expression in murine CD8 + T cells co‐cultured with mCAFs and iBMDMs ( n = 3 per group). P ‐values were calculated with one‐way ANOVA test with Tukey's post‐hoc test. ( B) In vivo bioluminescence imaging showing the growth of orthotopic lung tumors constructed by LLC, mCAFs‐vector/TDO2, and iBMDMs on day 21 ( n = 5 per group). ( C) Normalized bioluminescence imaging quantification of orthotopic lung tumors ( n = 5 per group). P ‐values were calculated with Student's t ‐test. ( D) Representative multiplexed IHC staining of orthotopic lung tumors stained for Pan‐CK, IL4I1, TDO2, F4/80, and COL1A1. Scale bar = 10 µm. ( E) Representative IHC staining of orthotopic lung tumors stained for Ki‐67, CD163, CD206, CD86, CD8, PD‐1, and GZMB. Scale bar = 50 µm. ( F) Schematic outline showing anti‐PD‐1 treatment for subcutaneous xenograft tumor formed by mouse KP lung cancer cells, mCAFs‐vector/TDO2, and iBMDMs in C57BL/6 mice. ( G) Gross appearance of subcutaneous xenograft tumor ( n = 5 per group). ( H) Tumor weight of subcutaneous xenograft tumor formed by mouse KP lung cancer cells, mCAFs‐vector/TDO2, and iBMDMs in the treatment of IgG2α and anti‐PD‐1 ( n = 5 per group). P ‐values were calculated with one‐way ANOVA test with Tukey's post‐hoc test. ( I) Bar plot showing the frequency of CD8 + GZMB + T cells, CD8 + PD‐1 + T cells among CD8A + T cell, and CD8 + TIM3 + T cells ( n = 5 per group). P ‐values were calculated with one‐way ANOVA test with Tukey's post‐hoc test. ( J) The concentrations of IL4I1 in the LUAD patients accepted anti‐PD‐1/PD‐L1 treatment. Violin plot showing the concentrations of blood plasma IL4I1 in PR ( n = 8) and SD/PD ( n = 11) LUAD patients (left). P ‐values were calculated with Student's t ‐test. Receiver operating characteristic (ROC) curves showing the diagnostic accuracy of IL4I1 (right) for prediction of immunology response.

Article Snippet: On day 9 post‐implantation, mice were randomly divided into four groups ( n = 5 per group) and intraperitoneally administered either IgG2α isotype control (BE0089, RRID: AB_1107769, BioXcell) or anti‐PD‐1 antibody (200 μg, BE0146, RRID: AB_10949053, BioXcell) every three days, corresponding to the following treatment groups: (1) mCAF‐vector + iBMDMs + IgG2α, (2) mCAF‐vector + iBMDMs + anti‐PD‐1, (3) mCAF‐TDO2 + iBMDMs + IgG2α, and (4) mCAF‐TDO2 + iBMDMs + anti‐PD‐1.

Techniques: Flow Cytometry, Expressing, Cell Culture, In Vivo, Imaging, Construct, Plasmid Preparation, Immunohistochemistry, Staining, Clinical Proteomics, Diagnostic Assay