Abstract 1546: Cortisol induces the expression of miR-143-3p/miR-145 cluster in macrophages to affect macrophage polarization via reprogramming of glucose metabolism

Abstract

Cortisol, the primary stress hormone, can contribute to cancer development through a variety of mechanisms. In a population-based research study, we have detected high serum levels of cortisol and several exosomal microRNAs (miRs) in women with a breast cancer (BC) diagnosis than non-cancer female subjects. In this study, we identified a positive correlation of cortisol levels with an exosomal miRNA, miR-143-3p, in serum samples. Furthermore, we observed an upregulation of miR-143-3p in human monocyte (THP1 and U937)-derived macrophages upon treatment with cortisol but not in BC cell lines (MDA-MB-468 and MDA-MB-231). Enhanced levels of miR-143-3p were also found in exosomes collected from the cortisol-treated macrophages. Expression of miR-145 (transcribed as cluster miRNA with miR-143-3p) was also upregulated in cortisol-treated macrophages but their enhanced levels were not detected in shed exosomes. In silico analysis identified glucocorticoid-response elements (GREs) in the upstream promoter sequence, which exhibited elicited transcriptional activity in response to cortisol in a luciferase-based promoter-reporter assay. Direct binding of glucocorticoid receptor (GR) to the regulatory sequence containing region was confirmed by chromatin immunoprecipitation. We further observed that cortisol treatment inhibited IFN-γ-induced M1 polarization, while caused alternative M2 polarization of macrophages and these effects were inhibited by pre-treatment with miR-143-3p and miR-145 inhibitors. Cortisol treatment reduced ECAR/OCR ratio in macrophages suggesting reduced glycolysis and/or increased oxidative phosphorylation. This effect was neutralized by functional inhibition of miR-143-3p and miR-145. Computational analysis using web-based servers (TargetScan and miRDB) identified HK2 (hexokinase-2) and ADPGK (ADP-dependent glucokinase) as potential targets of miR-143-3p and mir-145, respectively, whose expression was downregulated in cortisol-treated macrophages and rescued upon miR-143-3p and miR-145 inhibition. Direct targeting of 3′- untranslated regions of HK2 and ADPGK by miR-143-3p and miR-145, respectively, was confirmed in luciferase-based reporter assays. Altogether, we have identified novel cortisol-regulated miRNAs that inhibit classical M1 polarization and induce alternative M2 polarization by reprogramming of glucose metabolism. Our findings have relevance in establishing an indirect connection of chronic stress with cancer development via immunosuppressive effect of cortisol, and could be helpful in risk prediction and management efforts. Citation Format: Amod Sharma, Kunwar Somesh Vikramdeo, Sarabjeet Kour Sudan, Sachin Kumar Deshmukh, Ajay P. Singh, Seema Singh. Cortisol induces the expression of miR-143-3p/miR-145 cluster in macrophages to affect macrophage polarization via reprogramming of glucose metabolism [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1546.