The reduction of CSF1R-dependent CD115+Gr-1− blood monocytes served as a direct readout of the drug’s activity [22] (Supporting Information Fig. 14). Upon treatment, proliferation, as determined by the frequency of S phase cells, was completely blocked in both TAM subsets already at the earliest time point investigated (48 h) without significant rise in the apoptotic sub-G1 fraction (Fig. 6A). The check details continuous drug administration led to a drastic and persistent reduction of CD11bloF4/80hi TAMs (Fig. 6B) accompanied by a proliferation block and a twofold increase in apoptosis rate

(sub-G1; Fig. 6C). The abundance of the CD11bhiF4/80lo subset was not affected by GW2580 (Fig. 6B) and its proliferation and apoptosis remained unaltered at the later time points (Fig. 6C). These results reveal a crucial click here role of CSF1R in the maintenance or expansion of CD11bloF4/80hi

TAM, presumably through conveying prosurvival and/or growth signals. Since both STAT1 and CSF1R signaling fostered TAM accumulation in MMTVneu tumors, we explored the possibility of a mechanistic link between STAT1 and CSF1/CSF1R. Remarkably, significantly lower CSF1 amounts were detected in supernatants of Stat1-deficient primary tumor cultures (Fig. 7A) and in Stat1−/− tumor tissue (Fig. 7B) as compared with WT. Four putative STAT1-binding IFN-gamma activated sequence (GAS) elements were identified in the promoter of the mouse Csf1 gene in silico (Supporting Information Fig. 15A). Among them, GAS1 located in the first exon was bound by STAT1 in response to IFN-γ or IFN-γ/TNF-α stimulation in the MMTVneu tumor cells line (Fig. 7C and D). GAS1 exhibits a perfect homology across mammalian species, including the corresponding human sequence, described DOK2 to bind STAT1 in vitro [31] (Supporting Information Fig. 15B). Taken together, the interaction of STAT1 with the GAS1 element in the Csf1 promoter provides

a potential mechanistic basis for the heightened CSF1 levels and increased accumulation of CD11bloF4/80hi TAMs in Stat1-sufficient animals. Recent findings in the field of macrophage biology challenged the monocyte-centered view on the origin of mononuclear phagocytes. In particular, the discovery that a variety of tissue-resident macrophages self-maintain without contribution of circulating precursors [11-15] made us curious whether a similar mechanism accounts for TAM accumulation. We demonstrate here that local cell division of fully differentiated macrophages rather than low-pace supply of circulating monocytes fuels expansion of the predominant CD11bloF4/80hi TAM population in autochthonous HER2/NEU-driven neoplasms (Fig. 3, 4B, and 5). These findings contrast apparently with observations made in transplantation tumor models, where nondividing TAMs have been shown to arise solely from classical CD115+Ly6C+ blood monocytes [7, 20, 21, 27].