Journal: Journal for immunotherapy of cancer

Article Title: Targeting B7-H3 inhibition-induced activation of fatty acid synthesis boosts anti-B7-H3 immunotherapy in triple-negative breast cancer.

doi: 10.1136/jitc-2024-010924

Figure Lengend Snippet: Figure 1 B7-H3 is highly expressed in TNBC. (A) B7-H3 is highly expressed in BC tissues. Significance was calculated with the Wilcoxon test. (B) Single-cell RNA sequencing (scRNA-seq) analysis of 10 BC patients revealed the distribution of different cell types within the BC TIME and the correlation between B7-H3 expression and scRNA-seq distribution. (C) B7-H3 is highly expressed in TNBC tissues. Significance was calculated with the Wilcoxon test. (D) Kaplan-Meier plotter analysis showed that high B7-H3 expression is associated with poor outcomes in TNBC patients. (E, F) Representative images of B7-H3 immunohistochemical staining in TNBC and adjacent non-cancerous tissues from tissue microarrays. (G) Statistical analysis of the proportion of B7-H3-positive cells in TNBC versus adjacent tissues. (H, I) Western blot analysis showed that B7-H3 expression is significantly higher in TNBC cells than in non-cancerous breast epithelial cells. Significance was calculated using the Student’s t-test. ***p<0.001, ****p<0.0001. BC, breast cancer; TIME, tumor immune microenvironment; TNBC, triple-negative breast cancer.

Article Snippet: These agents included the B7- H3 mAb (BioXCell, Catalog BE0124), the FASN inhibitor (MCE, HY- 120394), and the PD- L1 mAb (BioXCell, Catalog BE0101).

Techniques: RNA Sequencing, Expressing, Immunohistochemical staining, Staining, Western Blot

Journal: Journal for immunotherapy of cancer

Article Title: Targeting B7-H3 inhibition-induced activation of fatty acid synthesis boosts anti-B7-H3 immunotherapy in triple-negative breast cancer.

doi: 10.1136/jitc-2024-010924

Figure Lengend Snippet: Figure 2 Biological functions of B7-H3 in TNBC cells. (A–C) Transfection efficiency of B7-H3 knockdown in TNBC cells. (D) B7-H3 knockdown suppressed TNBC cell proliferation as measured by the CCK-8 assay. (E, G) B7-H3 knockdown reduced colony formation capacity in TNBC cells. (F, H) B7-H3 knockdown impaired TNBC cell motility via transwell assay. Scale bars: 50 µm. (I, J) Downregulation of B7-H3 induced early apoptosis in TNBC cells. Data are presented as mean±SD (n=3). Significance was calculated using the Student’s t-test. **p<0.01, ***p<0.001, ****p<0.0001. TNBC, triple-negative breast cancer.

Article Snippet: These agents included the B7- H3 mAb (BioXCell, Catalog BE0124), the FASN inhibitor (MCE, HY- 120394), and the PD- L1 mAb (BioXCell, Catalog BE0101).

Techniques: Transfection, Knockdown, CCK-8 Assay, Transwell Assay

Journal: Journal for immunotherapy of cancer

Article Title: Targeting B7-H3 inhibition-induced activation of fatty acid synthesis boosts anti-B7-H3 immunotherapy in triple-negative breast cancer.

doi: 10.1136/jitc-2024-010924

Figure Lengend Snippet: Figure 3 Transcriptional and metabolomic profiling in B7-H3 knockdown TNBC cells. (A, B) GO enrichment analysis of differentially expressed genes related to biological processes. (C, D) GSEA plots illustrated significant enrichment of fatty acid beta-oxidation and fatty acid catabolic processes in the ranked gene dataset. (E) Heat map of the differentially expressed genes between si-NC and si-B7-H3 cells (n=3). (F) Western blot analysis showed that B7-H3 knockdown leads to an increase in FASN protein levels. (G) Schematic representation of metabolic enzymes and processes altered after B7-H3 knockdown in MDA-MB-231 cells. (H) MSEA illustrates the fold enrichment of various metabolic pathways, highlighting significant changes in metabolic activities. (I) Chord diagram displaying differentially expressed metabolites and their classifications. The different colors represent various metabolite classes, while the lines indicate Pearson correlation coefficients between metabolites, with red lines indicating positive correlations and blue lines indicating negative correlations. Fatty acid metabolic products are highlighted. Statistical significance was calculated with Student’s t-test. *p<0.05, **p<0.01, ****p<0.0001. MSEA, metabolite set enrichment analysis; TNBC, triple-negative breast cancer.

Article Snippet: These agents included the B7- H3 mAb (BioXCell, Catalog BE0124), the FASN inhibitor (MCE, HY- 120394), and the PD- L1 mAb (BioXCell, Catalog BE0101).

Techniques: Knockdown, Western Blot

Journal: Journal for immunotherapy of cancer

Article Title: Targeting B7-H3 inhibition-induced activation of fatty acid synthesis boosts anti-B7-H3 immunotherapy in triple-negative breast cancer.

doi: 10.1136/jitc-2024-010924

Figure Lengend Snippet: Figure 5 B7-H3 knockdown regulates FASN via the AKT pathway. (A) Visualization of TF family distribution by comparing all differentially expressed genes from transcriptomic data using BLAST against transcription factor databases. (B) Venn diagram showing candidate transcription factors selected from the JASPAR database and transcriptomic data. (C) Correlation between SREBP1 and FASN expression in TNBC. (D) The wild-type FASN promoter construct (pGL3-wt) exhibited significantly increased luciferase activity when cells were transfected with the pcDNA3.1 vector containing the SREBP1 gene. (E, F) B7-H3 knockdown upregulated SREBP1 expression. (G) Immunofluorescence assay showing nuclear translocation of SREBP1 following B7- H3 knockdown. Scale bars: 50 µm. (H, I) Western blot analysis confirmed the nuclear translocation of SREBP1 after B7-H3 knockdown. (J, K) Western blot analysis showed that protein levels of the Akt-mTOR signaling pathway were elevated in the si-B7-H3 group. (L–N) Western blot analysis demonstrating that inhibition of the Akt pathway suppressed the activation of both SREBP1 and FASN. Data are presented as mean±SD (n=3). Significance was calculated using the Student’s t-test. ns: non- significance, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: These agents included the B7- H3 mAb (BioXCell, Catalog BE0124), the FASN inhibitor (MCE, HY- 120394), and the PD- L1 mAb (BioXCell, Catalog BE0101).

Techniques: Knockdown, Expressing, Construct, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Immunofluorescence, Translocation Assay, Western Blot, Inhibition, Activation Assay

Journal: Journal for immunotherapy of cancer

Article Title: Targeting B7-H3 inhibition-induced activation of fatty acid synthesis boosts anti-B7-H3 immunotherapy in triple-negative breast cancer.

doi: 10.1136/jitc-2024-010924

Figure Lengend Snippet: Figure 6 Combined knockdown of B7-H3 and FASN more effectively suppresses TNBC cell growth. (A–C) Quantitative PCR and Western blot analyses confirmed the knockdown of B7-H3 and FASN at both mRNA and protein levels in MDA-MB-231 and BT-549 cells. (D, E) Combined knockdown of B7-H3 and FASN significantly inhibited colony formation capacity in TNBC cells. (F, G) Co-silencing of B7-H3 and FASN more effectively reduced TNBC cell migration. Scale bars: 50 µm. (H–I) Combined knockdown further suppressed the invasive ability of TNBC cells. Scale bars: 50 µm. (J–M) Co-knockdown of B7-H3 and FASN promoted early apoptosis in TNBC cells more significantly. Data presented as mean±SD (n=3). Statistical significance was calculated with one-way ANOVA with Tukey’s multiple-comparison test. ns: not significant, *p<0.05, ** p<0.01, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance; TNBC, triple-negative breast cancer.

Article Snippet: These agents included the B7- H3 mAb (BioXCell, Catalog BE0124), the FASN inhibitor (MCE, HY- 120394), and the PD- L1 mAb (BioXCell, Catalog BE0101).

Techniques: Knockdown, Real-time Polymerase Chain Reaction, Western Blot, Migration, Comparison

Journal: Journal for immunotherapy of cancer

Article Title: Targeting B7-H3 inhibition-induced activation of fatty acid synthesis boosts anti-B7-H3 immunotherapy in triple-negative breast cancer.

doi: 10.1136/jitc-2024-010924

Figure Lengend Snippet: Figure 7 Anti-B7-H3 and anti-FASN combination therapy enhances antitumor activity in vivo. (A) Schematic representation of the treatment protocol using anti-B7-H3, anti-FASN, anti-PD-L1, and combination therapy in BALB/c mice implanted with 4T1 cells (n=5). (B) Tumor growth curves showing tumor volume over time in each treatment group. (C) Comparative analysis of tumor weight among different treatment regimens. Mean±SD is shown (n=5). (D–E) Evaluation of FASN protein expression via Western Blot analysis. Anti-B7-H3 treatment significantly upregulated FASN compared with the control group. Mean±SD is shown (n=3). (F) Representative H&E staining of tumor sections. Scale bars: 50 µm. (G–H) TUNEL staining to assess apoptosis in tumor tissues. The combination therapy induced a higher level of apoptosis, as seen by the increased fluorescence ratio in H. Scale bars: 50 µm. Mean±SD is shown (n=3). (I–J) CD8 immunofluorescence staining to evaluate immune cell infiltration. The combination therapy group showed significantly enhanced CD8+ T cell infiltration, as quantified in panel I. Mean±SD is shown (n=3). (K) Visual pictures showing representative tumor volumes of each treatment group. (L) Tumor growth curves showing tumor volume in control, anti-PD-L1, and combination treatment groups. (M) Effect of anti-PD-L1 and combination therapy on tumor weight in balb/c mice bearing 4T1 cells. Mean±SD is shown (n=5). (N) Representative HE staining of tumor sections. Scale bars: 50 µm (left) and 20 µm (right). (O) TUNEL staining to assess apoptosis in tumor tissues. Scale bars: 50 µm (left) and 20 µm (right). (P) Immunofluorescence staining for CD8+ T Cell Infiltration in Tumor Section. Scale bars: 50 µm (left) and 20 µm (right). Statistical significance was calculated using the Student’s t-test for FASN expression and one-way ANOVA with Tukey’s multiple-comparison test for other comparisons. **p<0.01, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance.

Article Snippet: These agents included the B7- H3 mAb (BioXCell, Catalog BE0124), the FASN inhibitor (MCE, HY- 120394), and the PD- L1 mAb (BioXCell, Catalog BE0101).

Techniques: Activity Assay, In Vivo, Expressing, Western Blot, Control, Staining, TUNEL Assay, Fluorescence, Immunofluorescence, Comparison

Journal: Cell Death & Disease

Article Title: Oncogenic activation of SMYD3-SHCBP1 promotes breast cancer development and is coupled with resistance to immune therapy

doi: 10.1038/s41419-025-07570-8

Figure Lengend Snippet: Antibodies used for ChIP, IHC, IF, WB and in vivo mouse study.

Article Snippet: In Vivo MAb anti-mouse PD-1 (CD276) , BioX Cell , BE0146 , , .

Techniques: Immunohistochemistry-IF, In Vivo