Journal: Oncogene

Article Title: S100A8/A9 innate immune signaling as a distinct mechanism driving progression of smoking-related breast cancers.

doi: 10.1038/s41388-025-03276-5

Figure Lengend Snippet: Fig. 5 S100A8/A9 is required for the increased tumorigenicity due to chronic nicotine. A–C HCC38 tumorspheres after treatment with the indicated doses of the S100A8/A9 inhibitor paquinimod (ABR-215757) versus vehicle (DMSO) A, CRISPR/Cas9 knockout of S100A8 versus vector control B, or treatment with the RAGE inhibitor Azeliragon (TTP488) C. n = 3 independent experiments. D Representative images of tumors from HCC38 S100A8 KO or vector control cells treated with vehicle or chronic nicotine. Results are from orthotopic injection of 500 000 cells into inguinal mouse mammary glands and tumors harvested after 10 weeks. Scale bar, 1.5 cm. E Primary tumor volume versus days post- injection. D,E n = 16 tumors per cell type from 2 independent experiments. A–C,E Data represent the mean ± s.e.m. Statistics by two-way ANOVA and Tukey’s multiple comparisons test. A–C *P < 0.05 versus respective vehicle controls. n.s.=not significant. B *P < 0.05, ***P < 0.001. E *P < 0.05 for vehicle- versus nicotine-control cells and control- versus S100A8 KO-nicotine cells. See also Supplementary Fig. 5.

Article Snippet: For experiments with the S100A8/A9 inhibitor paquinimod (ABR-25757) or the RAGE inhibitor Azeliragon (TTP488) (Selleckchem, Houston, TX, USA), a single dose was added only once when embedding cells, and compared against cells receiving the same volume of vehicle (DMSO).

Techniques: CRISPR, Knock-Out, Plasmid Preparation, Control, Injection

Journal: NPJ Breast Cancer

Article Title: RAGE inhibitor TTP488 (Azeliragon) suppresses metastasis in triple-negative breast cancer

doi: 10.1038/s41523-023-00564-9

Figure Lengend Snippet: a 4175 cells were injected into the mammary fat pad of immunocompromised NSG mice, and mice were treated IP twice per week with 1 mg/kg TTP488 or FPS-ZM1 (or vehicle (DMSO) control). Tumor size was measured every 3 days, and ( b ) at the end of the experiment, tumors were weighed. Data shown are from 4–8 mice per group (repeated twice). c Immunohistochemical analysis of 4175/NSG tumors for proliferation (Ki67). d Immunohistochemical analysis of human CK7 to assess for metastasis in 4175/NSG mouse lung tissue. e 4T-1 cells were tail-vein injected into BALBc mice, and mice were treated IP twice per week with 1 mg/kg TTP488 or FPS-ZM1 (or vehicle (DMSO) control). Representative IVIS images at day 13 are shown from n = 11 per group. f Total flux measured from IVIS images is shown. G Surface lung metastases count performed at necropsy. Values shown are mean ± SD. Statistical analysis: one or two-way ANOVA with Dunnett’s multiple comparisons test. * p < 0.05; ** p < 0.01; *** p < 0.001, **** p < 0.0001.

Article Snippet: Most importantly, TTP488 (developed by Transtech Pharma, now vTv Therapeutics) advanced to clinical trials and showed a favorable safety profile at 5 mg taken once daily in clinical trials of patients with Phase 2 mild Alzheimer’s disease .

Techniques: Injection, Control, Immunohistochemical staining

Journal: NPJ Breast Cancer

Article Title: RAGE inhibitor TTP488 (Azeliragon) suppresses metastasis in triple-negative breast cancer

doi: 10.1038/s41523-023-00564-9

Figure Lengend Snippet: a Venn-diagram of the overlap in Differentially Expressed Genes (DEGs) in the tumors of the two treatment groups (TTP488 and FPS-ZM1). b Scatterplot of the log2-fold change of DEGs calculated in one or both treatments relative to DMSO control in primary tumors. A trend line of plotted genes (red) juxtaposes a reference (dotted diagonal) line denoting equivocal expression fold change by both treatments. c Venn diagram of the overlap in DEGs in the TTP488 and FPS-ZM1 treated lungs. d Scatterplot of the log2 fold change of DEGs calculated in one or both treatments relative to DMSO control in metastatic lungs. e , f Scatterplot of the enriched terms in the KEGG 2021 Human library in tumor ( e ) and lung ( f ) for TTP488 treated mice versus vehicle control. Significantly enriched pathway terms overlapping between the primary tumor and the pulmonary metastasis are marked with gold circles and labels; non-overlapping significantly enriched terms are marked and labeled in black. g , h Bar graphs show the top 20 KEGG and GO BP cancer-associated enriched pathway terms in order from top to bottom by the magnitude of significance for genes significantly changed with TTP488 treatment relative to DMSO control in tumor versus lung metastasis.

Article Snippet: Most importantly, TTP488 (developed by Transtech Pharma, now vTv Therapeutics) advanced to clinical trials and showed a favorable safety profile at 5 mg taken once daily in clinical trials of patients with Phase 2 mild Alzheimer’s disease .

Techniques: Control, Expressing, Labeling

Journal: NPJ Breast Cancer

Article Title: RAGE inhibitor TTP488 (Azeliragon) suppresses metastasis in triple-negative breast cancer

doi: 10.1038/s41523-023-00564-9

Figure Lengend Snippet: a Cell adhesion to various extracellular matrix (ECM) component proteins was assessed using the ECM Cell Adhesion Array. 4175 cells were treated with TTP488 (1 µM) or vehicle control for 48 h, before adhesion assays. 4175 cells (150,000 cells per well) were incubated for 2 h at 37 °C in the ECM Array. Attached cells were stained with crystal violet and quantified. Data shows two experimental repeats run in triplicate. b , c Boyden chamber invasion assay of 4175 and 4T1 cells treated with FPS-ZM1 (1 µM) and TTP488 (1 µM). d , e Boyden chamber migration assay of 4175 and 4T1 cells treated with FPS-ZM1 (1 µM) and TTP488 (1 µM). Values show mean + SD from three independent experiments performed in triplicate. Statistical analysis was performed with either one-way or two-way ANOVA. ** p < 0.01; *** p < 0.001, **** p < 0.0001.

Article Snippet: Most importantly, TTP488 (developed by Transtech Pharma, now vTv Therapeutics) advanced to clinical trials and showed a favorable safety profile at 5 mg taken once daily in clinical trials of patients with Phase 2 mild Alzheimer’s disease .

Techniques: Control, Incubation, Staining, Invasion Assay, Migration

Journal: NPJ Breast Cancer

Article Title: RAGE inhibitor TTP488 (Azeliragon) suppresses metastasis in triple-negative breast cancer

doi: 10.1038/s41523-023-00564-9

Figure Lengend Snippet: a – d Cell viability/proliferation assays were performed using the crystal violet assay. 4175 and 4T1 cells were treated with a range of concentrations (50–1000 nM) of FPS-ZM1 or TTP488 for 24, 48, or 72 h. a 4175 cell treatment with FPS-ZM1; ( b ) 4T1 treatment with FPS-ZM1; ( c ) 4175 treatment with TTP488; ( d ) 4T1 treatment with TTP488. e Cell cycle analysis of 4175 cells treated in vitro with 1000 nM of FPS-ZM1 or TTP488 for 24, 48, or 72 h. Cells were stained with propidium iodide solution and analyzed for cell cycle distribution by flow cytometry. Representative column graph showing the percentage of cells in each cell cycle. Values show mean + SD from 3–5 independent experiments performed in triplicate. Statistical analysis was performed with either one-way or two-way ANOVA. * p < 0.05.

Article Snippet: Most importantly, TTP488 (developed by Transtech Pharma, now vTv Therapeutics) advanced to clinical trials and showed a favorable safety profile at 5 mg taken once daily in clinical trials of patients with Phase 2 mild Alzheimer’s disease .

Techniques: Crystal Violet Assay, Cell Cycle Assay, In Vitro, Staining, Flow Cytometry

Journal: NPJ Breast Cancer

Article Title: RAGE inhibitor TTP488 (Azeliragon) suppresses metastasis in triple-negative breast cancer

doi: 10.1038/s41523-023-00564-9

Figure Lengend Snippet: a , b Serum from 4175 tumor-bearing mice was assessed for changes in human tumor-intrinsic systemic changes due to RAGE inhibition by protein arrays. 3 pooled samples (for each of DMSO, FPS-ZM1 and TTP488) were assessed by the Proteome Profiler Human XL cytokine array; error bars represent two technical repeats per analyte. a All proteins that displayed differences compared to DMSO control by protein array (Proteome Profiler Human XL cytokine array) are shown. Data is shown as an average of mean pixel density for each pair of duplicate spots for each cytokine. b Log2-fold change in serum protein differences due to RAGE inhibitors relative to DMSO control. c GM-CSF ELISA was performed on serum from 4175 tumor-bearing mice. Samples (four per group) were run in duplicates for each individual sample. d IL8 ELISA was performed on serum from 4175 tumor-bearing mice. Samples (four per group) were run in duplicates for each individual sample. e Clustergram (of MsigDB Hallmark pathways) analysis generated by ENRICHR of tumor-derived protein changes in mouse serum.

Article Snippet: Most importantly, TTP488 (developed by Transtech Pharma, now vTv Therapeutics) advanced to clinical trials and showed a favorable safety profile at 5 mg taken once daily in clinical trials of patients with Phase 2 mild Alzheimer’s disease .

Techniques: Inhibition, Control, Protein Array, Enzyme-linked Immunosorbent Assay, Generated, Derivative Assay

Journal: NPJ Breast Cancer

Article Title: RAGE inhibitor TTP488 (Azeliragon) suppresses metastasis in triple-negative breast cancer

doi: 10.1038/s41523-023-00564-9

Figure Lengend Snippet: a , b Tumor lysate from 4175 tumor-bearing mice was assessed for changes in signaling pathway mechanisms due to RAGE inhibition by phospho-protein arrays. 3 pooled samples (for each of DMSO and TTP488) were assessed by the Proteome Profiler Human Phospho-Kinase Array Kit. a All phospho-proteins that displayed differences compared to DMSO control by protein array are shown. Data is shown as average of mean pixel density for each pair of duplicate spots for each cytokine. b Log2-fold change in phospho-protein differences due to RAGE inhibitor TTP488 relative to DMSO control. c Western blot validation of phospho-protein array changes was performed with tumor lysate from 4175 tumor-bearing mice (4 samples per condition). Representative images are shown for each condition. Samples were analyzed for phospho and total protein for Pyk2, STAT3 and AKT, and beta-actin loading control. d RAGE signaling in TNBC drives tumor metastasis. A schematic depicting the major signaling pathways activated by RAGE in TNBC cells leading to metastasis.

Article Snippet: Most importantly, TTP488 (developed by Transtech Pharma, now vTv Therapeutics) advanced to clinical trials and showed a favorable safety profile at 5 mg taken once daily in clinical trials of patients with Phase 2 mild Alzheimer’s disease .

Techniques: Inhibition, Control, Protein Array, Western Blot, Biomarker Discovery, Protein-Protein interactions