injection with trametinib  (TargetMol)

Journal: iScience

Article Title: Tracing specificity of immune landscape remodeling associated with distinct anticancer treatments

doi: 10.1016/j.isci.2025.112071

Figure Lengend Snippet:

Article Snippet: Another cohort was treated for 10 or 20 days, every day by intraperitoneal injection with Trametinib (1 mg/kg, MEK inhibitor; TargetMol) plus ABT-737 (30 mg/kg, BCL-XL inhibitor; TargetMol), both diluted in Cremophore and NaCl 0.9%.

Techniques: Recombinant, Software

Journal: Cancer Discovery

Article Title: Early Infiltration of Innate Immune Cells to the Liver Depletes HNF4α and Promotes Extrahepatic Carcinogenesis

doi: 10.1158/2159-8290.CD-22-1062

Figure Lengend Snippet: Breast cancers induce metabolic changes in the liver during early carcinogenesis. A, RT-PCR of livers from breast cancer–bearing mice demonstrates decreased RNA expression of UC enzymes along cancer progression ( n = 5, Student t test). Day 4, P = 0.003, day 14, P = 0.033, 0.01, and 0.028 (respectively), day 21, P = 0.0002, 0.019, 0.0002, 0.013, and 0.007 (respectively). B, Left, Western blots demonstrating decreased protein expression levels of ASS1 and OTC in livers of breast cancer–bearing mice vs. WT PBS-injected mice ( n = 5, Student t test). Right, quantification of band intensities: ASS1, P = 0.002; OTC, P = 0.011. C, RT-PCR of liver from MMTV-PyMT of 14-week-old cancer-bearing mice demonstrate decreased RNA expression of UC enzymes compared with livers of WT mice (WT n = 5, MMTV-PyMT n = 3); P = 0.033, 3.14E−05, 0.038 (respectively). D, Measurements of UC-related metabolites following glutamine 15 N 2 infusion to 4T1 breast cancer–bearing or PBS-injected mice in the plasma and livers. Left, a decreased ratio of urea to glutamine and glutamate m + 1 isotopologues in the plasma of 4T1 breast cancer–bearing mice (WT n = 5, 4T1 n = 7, Student t test); P = 0.019, 0.042, respectively. Right, an increase in the levels of glutamate (AUC/internal standard/dry weight) in 4T1 breast cancer–bearing mice and a decrease in the ratio of urea to glutamate following infusion in 4T1 breast cancer–bearing mice, supporting a malfunctioning UC ( n = 7, Student t test); P = 0.016, 0.048, respectively. E, Left, measurements of ammonia levels, using an ammonia assay kit, in the plasma of breast cancer–bearing mice compared with WT PBS-injected mice (WT n = 4, an outlier – interquartile range method, 4T1 n = 5, Student t test). Right, urea levels in the urine of breast cancer–bearing mice compared with WT PBS-injected mice. P = 0.033. F, Left, decreased levels of m + 1–labeled aspartate in the tumor of 4T1 breast cancer–bearing mice compared with the levels in the plasma and liver following infusion of 15 N 2 glutamine ( n = 8, plasma n = 7, two-way ANOVA); P values: plasma vs. tumor = 0.041, liver vs. tumor = 0.007. Right, increased ratio of uracil m + 1 to aspartate (Asp) m + 1 isotopologues in the tumors of 4T1 breast cancer–bearing mice ( n = 8, plasma n = 7, two-way ANOVA); P values: plasma vs. tumor = 0.042, liver vs. tumor = 0.008. G, XTT assay for 4T1 cancer cell proliferation following supplementation of the medium with UC intermediates ( n = 3, two-way ANOVA). P values: 24 hours: aspartate: not significant, ammonia = 0.007, glutamate and fumarate <0.0001; 48 and 72 <0.0001 for all measurements. Metabolite concentrations were supplemented in the following concentrations: ammonia, 0.75 mmol/L as published ; aspartate, 0.25 mmol/L; fumarate, 0.35 mmol/L; glutamate, 0.25 mmol/L. H, Ex vivo FACS analysis of CD8 + splenocyte cell survival (left) and activation (right) following supplementation with 1 mmol/L ammonia (Amn; ref. ; measured plasma ammonia levels 1.34 mmol/L; n = 5, Student t test); P = 0.011, 0.043, respectively. I, Heat map for differential gene expression in hepatocytes demonstrates a unique pattern on day 21 in the livers of 4T1 breast cancer mice compared with day 4. J, 4T1 day 21 vs. 4T1 day 4 pathway enrichment analysis. Each bar shows the fold enrichment of a specific pathway. TCA, tricarboxylic acid.

Article Snippet: Following 24 hours of 4T1 breast cancer cell injection, mice were injected i.p. with the 1 mg/kg of the ERK inhibitor trametinib (GSK1120212; Selleckchem; #S2673) in 4% DMSO corn oil or 4% DMSO corn oil only 6 more times a week.

Techniques: Reverse Transcription Polymerase Chain Reaction, RNA Expression, Western Blot, Expressing, Injection, Clinical Proteomics, Labeling, XTT Assay, Ex Vivo, Activation Assay, Gene Expression

Journal: Cancer Discovery

Article Title: Early Infiltration of Innate Immune Cells to the Liver Depletes HNF4α and Promotes Extrahepatic Carcinogenesis

doi: 10.1158/2159-8290.CD-22-1062

Figure Lengend Snippet: Innate immune cells infiltrate the host's liver during early breast carcinogenesis. A, Hematoxylin and eosin staining of liver sections demonstrates increased immune cell infiltration along a time course following breast cancer cell injection. The black arrows point to the infiltrating immune cells. Magnifications: 10 and 40× as detailed in the figure. B, Cell types annotated from the scRNA-seq analysis are projected on three uniform manifold approximation and projections (UMAP), indicating which cells appear at which time point. The arrows mark the neutrophil and monocyte subsets that were absent in the liver of WT mice and gradually accumulated in the liver until day 21. DC, dendritic cell; moDC, monocyte-derived dendritic cell; NK, natural killer; pDC, plasmacytoid dendritic cell; Treg, regulatory T cell. C–E, CyTOF-t-distributed stochastic neighbor embedding (t-SNE) analysis ( C and E ) and ( D ) quantification of liver and blood CD45 + populations show increased levels of innate immune cells and reduction of lymphocytes in the liver ( C ) and blood ( E ) of breast cancer–bearing mice compared with WT PBS-injected mice (WT n = 3, 4T1 = 5, Student t test). P values for blood <0.0001. P values for liver: B cells = 0.013, T cells = 0.0014, neutrophils = 0.0001. F, Multiplex ELISA immunoassay demonstrates increasing levels of IL6 (left) and TNFα (right) in breast cancer–bearing mice from day 4 to day 21 following cancer cell injection ( n = 5, Student t test). P values for IL6: day 4 = 0.024, day 14 <0.0001, and day 21 = 0.0009. P values for TNFα: day 4 = 0.05, day 14, and day 21 <0.0001. G, ELISA assay measurements of CCL2 levels in plasma, livers, spleens, and lungs of breast cancer–bearing mice ( n = 5, Student t test). P values: plasma = 0.0005, liver = 0.017, spleen = 0.04. H, ELISA assay demonstrates a significant interaction effect between the time following cancer cell injection and elevation of CCL2 in the liver of breast cancer–bearing mice relative to WT PBS-injected mice ( n = 5, two-way ANOVA); P = 0.0034. I, Expression of CCR2 in immune cells from the scRNA-seq experiment, projected on the UMAP, demonstrates that CCR2 is mainly expressed in monocyte subpopulations.

Article Snippet: Following 24 hours of 4T1 breast cancer cell injection, mice were injected i.p. with the 1 mg/kg of the ERK inhibitor trametinib (GSK1120212; Selleckchem; #S2673) in 4% DMSO corn oil or 4% DMSO corn oil only 6 more times a week.

Techniques: Staining, Injection, Derivative Assay, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Expressing

Journal: Cancer Discovery

Article Title: Early Infiltration of Innate Immune Cells to the Liver Depletes HNF4α and Promotes Extrahepatic Carcinogenesis

doi: 10.1158/2159-8290.CD-22-1062

Figure Lengend Snippet: IL6–STAT3–HNF4α signaling causes changes in liver metabolism during breast cancer carcinogenesis. A, Differentially expressed gene pathway enrichment analysis for genes regulated by HNF4α (Supplementary Table S1) in livers of breast cancer–bearing mice on day 21 compared with day 4. The bar shows the Z score of a specific pathway. B, Left, plasma albumin (Alb) measurements demonstrate decreased levels in breast cancer–bearing mice compared with WT PBS-injected mice ( n = 5, Student t test); P = 0.002. Right, normalized RNA-seq analysis of albumin expression in hepatocytes on days 4 and 21. P = 0.013. C, RT-PCR of primary hepatocytes demonstrates decreased RNA expression levels of Otc following IL6 supplementation ( n = 3, Student t test, representative experiment of two independent biological replicates); P = 0.0002. D, STAT3 inhibitor HJCO152 restores Otc levels even in the presence of IL6 ( n = 3, one-way ANOVA); P < 0.0001 for CTRL + DMSO vs. IL6 + DMSO and STAT3 inhibitor vs. STAT3 inhibitor + IL6. inh, CTRL + DMSO vs. STAT3 inh + IL6; ns, not significant. E, Western blots (left) demonstrate increased expression levels of HNF4α in breast cancer–bearing mice treated with an ERK inhibitor compared with livers from breast cancer–bearing mice treated with vehicle (vehicle n = 4, trametinib n = 3, Student t test). Right, quantification of band intensities. P = 0.024. F, Measurement of tumor weight on days 8 and 14 following treatment with an ERK inhibitor (day 8 n = 5, day 14 n = 4, Student t test). P values: day 8 = 0.007 and day 14 = 0.014. G, RT-PCR of livers from breast cancer–bearing mice treated with AAV8-HNF4α demonstrates increased RNA expression of Hnf4 a, UC enzymes, and Alb in comparison with AAV8-GFP–treated breast cancer–bearing mice (AAV8-GFP n = 4, AAV8-HNF4α n = 7, Student t test); P = 0.004, 0.004, 0.002, 0.001, respectively. H, 4T1 tumor growth is significantly higher following AAV-GFP injection compared with AAV-HNF4α (AAV8-GFP n = 12, AAV8-HNF4α n = 13, Student t test). P = 0.049. I, Left, Western blots showing decreased levels of PCNA in tumors from breast cancer–bearing mice treated with HNF4α compared with mice treated with GFP. Right, quantification of band intensity relative to actin. P = 0.003.

Article Snippet: Following 24 hours of 4T1 breast cancer cell injection, mice were injected i.p. with the 1 mg/kg of the ERK inhibitor trametinib (GSK1120212; Selleckchem; #S2673) in 4% DMSO corn oil or 4% DMSO corn oil only 6 more times a week.

Techniques: Clinical Proteomics, Injection, RNA Sequencing, Expressing, Reverse Transcription Polymerase Chain Reaction, RNA Expression, Western Blot, Comparison