Figure 1.

TRIB1 is associated with overall BC survival and 5-years relapse-free cancer survival after chemotherapy and is a highly expressed protein in Tumor-Associated Macrophages. (A) TRIB1 mutation-harboring tumors are associated with a poor long-term BC survival. Kapplan-Maier survival plot, Hazard Ratio (HR) and log rank p value are displayed. Red line represents patients harboring the transcriptomic fingerprint of TRIB1 somatic mutants vs. wild type TRIB represented by a black line. (B) TRIB1 expression in 5-year relapse-free BC survival after anthracycline-based chemotherapy. Note that enhanced TRIB1 expression correlates with a higher 5-year relapse-free BC survival after anthracycline-based chemotherapy. Receiver Operating characteristic (ROC) curves are shown. Area under the curve (AUC), p value, false positive rate (FPR) and true positive rate (TPR). (B) All breast cancer patients, (C) basal-like and (D) luminal-B subtypes. (E) Representative image of macrophages in human breast cancer marked with CD68 (red) and TRIB1 (green) (Scale: 100 µm). Quantification of TRIB1 expressing cells and CD68+ cells in the TMA from “E” relative to total cell counts. Data presented as mean±SD. Cells in 5 random fields of views for each tumor were manually quantified using ImageJ. (F) Representative image of TRIB1 (red) and F4/80 (green) fluorescence staining of primary murine breast tumors from wild-type (C57/BL6) animals (Scale: 50 µm). Quantification of TRIB1 expressing cells and F4/80+ cells in the TME from “F” relative to total cell counts (n=5 mice/group). Data presented as mean±SEM. Cells in 5 random fields of views for each animal were manually quantified using ImageJ. (G) Human MDMs isolated from healthy donors were transfected with either control (M^Control) or TRIB1 siRNA for 48 hours (M^TRIB1-KD). Results of paired t-tests are presented; mean±SEM is plotted; *p < 0.05, **p < 0.01 (n= 6-9 donor/group). (H) Pathway analysis of TRIB1 associated genes in monocytes and MDMs in participants of the Cardiogenics Transcriptomic Study. MDM (n= 596) and monocytes (n= 758) were ranked according to TRIB1 RNA levels and genes that were differentially expressed between the top vs. bottom 25% of the rankings were analyzed with QuSAGE. The 10 most significantly enriched pathways in MDMs are presented. (I) TRIB1 RNA levels of human MDMs unstimulated (M) and stimulated with LPS and INF-γ (M^LPS+INF-γ), IL-4 (M^IL-4), and CM (M^CM) for 24 hours. Results of Friedman test and Dunn's multiple comparison tests are presented; mean±SEM is plotted; *p < 0.05 (n = 6 donors/group).