Fig. 4: STING agonists can reprogram M2-like TEMs/TAMs into an M1-like state. | Nature Communications

Fig. 4: STING agonists can reprogram M2-like TEMs/TAMs into an M1-like state.

From: STING agonism reprograms tumor-associated macrophages and overcomes resistance to PARP inhibition in BRCA1-deficient models of breast cancer

Fig. 4: STING agonists can reprogram M2-like TEMs/TAMs into an M1-like state.The alternative text for this image may have been generated using AI.

a and b Mouse BMDMs were subjected to transcriptome analysis after treatment for 24 h with DMSO vehicle control, olaparib (OL, 5 μM), DMXAA (0.05 mg/mL), 50% BP-CM with or without DMXAA, or 50% BP/OL-CM with or without DMXAA. a Heat map of anti-tumor and pro-tumorigenic gene expression in BMDMs (n = 2 for each group). b Left, volcano plot showing the significance and magnitude of changes in gene expression of BMDMs treated with BP-CM/DMXAA compared to BP-CM/DMSO (n = 2 for each group). Statistical significance was calculated using a two-sided Wald test and adjusted for multiple testing using the Benjamini–Hochberg procedure. Right, top-ranked up-regulated and down-regulated gene ontology (GO) terms in BMDMs treated with BP-CM/DMXAA. Significance of enriched terms were adjusted using the Benjamini–Hochberg procedure for multi-testing. c Analysis of control macrophages (Mφ) and BP TEMs treated with or without DMXAA (0.05 mg/mL) for 2 days. M1-like (CD11b+ CD206- MHC-IIhigh) to M2-like (CD11b+ CD206+ MHC-IIlow) ratio was analyzed by flow cytometry (n = 4). d Analysis of M1 to M2 ratio of TAMs (sorted from BP tumors) treated with or without DMXAA (0.05 mg/mL) for 2 days (n = 3). e Diagram of workflow for (f-h). BMDMs isolated from wildtype (WT) or STING knockout (STING-/-) C57/BL6J mice were incubated with control medium or 50% BP-CM to generate control naïve BMDMs and TEMs, respectively. Cells were then treated with or without DMXAA (0.05 mg/mL) for 2 days. f and g M1 to M2 ratio (f) and surface levels of the co-stimulatory molecule CD86 (g) were analyzed by flow cytometry (n = 4). h BP cells were co-cultured with or without WT TEMs or STING−/− TEMs pre-treated with or without DMXAA (0.05 mg/mL), followed by 2 days of olaparib (5 μM) treatment. BP cells were then analyzed for apoptosis (Annexin V+ 7-AAD−; BP cells, n = 6; BP + WT TEMs, n = 3; BP + STING−/− TEMs, n = 3). i Expression of M2 (CD163) and M1 (CD86) markers in control THP-1 macrophages or MDA-MB-436 tumor cell-educated THP-1 macrophages (TEMs-436) treated with or without ADU-S100 (10 μM) for two days (n = 3). Data are presented as mean ± SEM. One-way ANOVA (c) and (f)–(i). Two-tailed unpaired t test (d). ns, not significant. Source data are provided as a Source Data file.

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