Journal: Advanced Science

Article Title: Three‐Year Follow‐Up of Neoadjuvant Tislelizumab with Chemotherapy in Locally Advanced Gastric and Gastroesophageal Junction Adenocarcinoma: Revealing Cancer‐Associated Fibroblast Heterogeneity Corresponding to PD‐1 Blockade Efficacy

doi: 10.1002/advs.202508433

Figure Lengend Snippet: eCAFs and iCAFs showed different reactions to immunotherapy. a) UMAP visualization of CAF subtypes identified in LAGC tissues. b) Proportions of CAF subtypes in responders and non‐responders. c) UMAP feature plots showing the expression levels of representative marker genes (ACTA2, COL14A1, FAP, and SLIT2) in CAF subtypes. d) GO functional enrichment analysis of iCAFs, eCAFs, and myCAFs. e) KEGG pathway enrichment analyses of iCAFs, eCAFs, and myCAFs. f) GSEA of iCAFs, eCAFs, and myCAFs. GO, gene ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; GSEA, gene set enrichment analysis; CAF, cancer‐associated fibroblast; iCAF, inflammatory CAF; eCAF, extracellular matrix‐related CAF; myCAF, myofibroblastic CAF; LAGC, locally advanced gastric cancer.

Article Snippet: Multiplex immunohistochemical (IHC) staining was performed using the Opal 5‐color kit (Akoya Bioscience, NEL801001KT) to evaluate CFD (GB12045, Servicebio, 1:5000, Opal 440), COL14A1 (AF0573, Affinity Biosciences, 1:2000, Opal 488), FAP (11779‐1‐AP, 1:2000, Opal 555), SLIT2 (20217‐1‐AP, ProteinTech, 1:2000, Opal 647), CXCL14 (10468‐1‐AP, ProteinTech, 1:1000, Opal 647) and POSTN (66491‐1‐Ig, 1:1000, Opal 488).

Techniques: Expressing, Marker, Functional Assay

Journal: Advanced Science

Article Title: Three‐Year Follow‐Up of Neoadjuvant Tislelizumab with Chemotherapy in Locally Advanced Gastric and Gastroesophageal Junction Adenocarcinoma: Revealing Cancer‐Associated Fibroblast Heterogeneity Corresponding to PD‐1 Blockade Efficacy

doi: 10.1002/advs.202508433

Figure Lengend Snippet: The different CAF subtypes involved the immune microenvironment. a) UMAP of all spots (n = 29808) from nine GC primary samples. b) The proportion of cell types in all samples. c) Violin plots showing the expression of immune‐stimulating factors CXCL8, CXCL9, CXCL10, CXCL11, CCL4, CCL5, and IFNG. d) Heatmap for AUcell scores of immune pathways and CAF subtypes (iCAFs and eCAFs) across GC samples. e) The spatial representation of cell type niches in STAD#2 (immunogenic GC) and STAD#5 (non‐immunogenic GC). f) The expression level of iCAFs in spots of STAD#2 and STAD#5. g) The spatial representation of iCAFs in spots of STAD#2 and STAD#5. h) Violin plots for the expression of POSTN, CXCL14, CFD, CD34, SLIT2, FAP, and ACTA2 in all GC samples. i) The spatial representation of CD8+ T cells in spots of STAD#2, STAD#8, (immunogenic GC) STAD#5, and STAD#6 (non‐immunogenic GC). * p < 0.05, ** p < 0.01, *** p < 0.001. CAF, cancer‐associated fibroblast; iCAF, inflammatory CAF; eCAF, extracellular matrix‐related CAF; GC, gastric cancer.

Article Snippet: Multiplex immunohistochemical (IHC) staining was performed using the Opal 5‐color kit (Akoya Bioscience, NEL801001KT) to evaluate CFD (GB12045, Servicebio, 1:5000, Opal 440), COL14A1 (AF0573, Affinity Biosciences, 1:2000, Opal 488), FAP (11779‐1‐AP, 1:2000, Opal 555), SLIT2 (20217‐1‐AP, ProteinTech, 1:2000, Opal 647), CXCL14 (10468‐1‐AP, ProteinTech, 1:1000, Opal 647) and POSTN (66491‐1‐Ig, 1:1000, Opal 488).

Techniques: Expressing

Journal: Advanced Science

Article Title: Three‐Year Follow‐Up of Neoadjuvant Tislelizumab with Chemotherapy in Locally Advanced Gastric and Gastroesophageal Junction Adenocarcinoma: Revealing Cancer‐Associated Fibroblast Heterogeneity Corresponding to PD‐1 Blockade Efficacy

doi: 10.1002/advs.202508433

Figure Lengend Snippet: Multi‐IF confirmed the duality of CAFs in immunotherapy. a,b) Multiplexed immunofluorescence staining of FAP, SLIT2, CFD, and COL14A1 in LAGC tissues of responders and non‐responders treated with immunotherapy. Multiplexed immunofluorescence assays were performed twice on tumor samples following assay optimization. c–f) Quantification of PCP for COL14A1, FAP, SLIT2, and CFD in patients with LAGC stratified by TRGs. g) Kaplan‐Meier analysis of PFS based on different expression levels of COL14A1, FAP, SLIT2, and CFD in patients receiving immunotherapy. h) Kaplan‐Meier analysis of OS according to the expression levels of COL14A1, FAP, SLIT2, and CFD in patients receiving immunotherapy. PCP, percentage of positive cells; TRG 0, (complete pathological response in the primary tumor and lymph nodes, no residual tumor cells), TRG 1, (near‐complete response, ≤10% residual tumor cells), TRG 2, (partial response, 10–50% residual tumor cells), and TRG 3, (poor or no response, ≥50% residual tumor cells). PFS, prognosis‐free survival; OS, overall survival; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Multiplex immunohistochemical (IHC) staining was performed using the Opal 5‐color kit (Akoya Bioscience, NEL801001KT) to evaluate CFD (GB12045, Servicebio, 1:5000, Opal 440), COL14A1 (AF0573, Affinity Biosciences, 1:2000, Opal 488), FAP (11779‐1‐AP, 1:2000, Opal 555), SLIT2 (20217‐1‐AP, ProteinTech, 1:2000, Opal 647), CXCL14 (10468‐1‐AP, ProteinTech, 1:1000, Opal 647) and POSTN (66491‐1‐Ig, 1:1000, Opal 488).

Techniques: Immunofluorescence, Staining, Expressing