Tumor-Expressed BMAL1 Influences Immune Cell Infiltration
(A) F4/80 signal was quantified as the average area of F4/80 staining in non-overlapping sections of entire tumor and compared with other samples at the same magnification. n = 4 (left panel). Representative images of tissue sections are shown (right panel). Scale bar, 500 μm. Red scale bar, 125 μm (inset).
(B) CD8 signal was quantified as the average area of CD8 staining in non-overlapping sections of entire tumor section and compared with other samples at the same magnification. n = 4. (A and B) Data shown as mean ± SEM; *p < 0.05; **p < 0.01; one-way ANOVA with Tukey's post hoc. Representative images of stained tissue are shown (right panel). Scale bar, 250 μm. Red scale bar, 100 μm (inset).
(C) Gene Set Enrichment Analysis shows that expression of hallmark OxPhos gene set in TCGA BC primary tumors (n = 1,097) is inversely correlated with BMAL1 expression.
(D and E) Kaplan-Meier curves show distal metastasis-free survival rate of patients selected according to BMAL1 expression from two independent datasets GEO: GSE20685 (D) and GSE11121 (E).
(F) Hyperinsulinemia and BMAL1 regulate both internal metabolism and external microenvironment of TNBC. The intrinsic metabolism of TNBC reveals that BMAL1 and hyperinsulinemia have parallel yet distinct regulations of pyruvate and mitochondrial metabolism. The external TNBC microenvironment shows that BMAL1 and hyperinsulinemia in concert control immune cell recruitment and infiltration. The dotted line indicates a possible mechanism for immune cell recruitment via mitochondria-induced reactive oxygen species.