Sustained inhibition of CSF1R signaling augments antitumor immunity through inhibiting tumor-associated macrophages

Tumor-associated macrophages (TAMs) are one of the key immunosuppressive components in the tumor microenvironment (TME) and contribute to tumor development, progression, and resistance to cancer immunotherapy. Several reagents targeting TAMs have been tested in preclinical and clinical studies, but they have had limited success. Here, we show that a unique reagent, FF-10101, exhibits a sustained inhibitory effect against colony stimulating factor 1 receptor by forming a covalent bond and reduces immunosuppressive TAMs in the TME, which leads to strong antitumor immunity. In preclinical animal models, FF-10101 treatment significantly reduced immunosuppressive TAMs and increased antitumor TAMs in the TME. In addition, tumor antigen-specific CD8+ T cells were increased; consequently, tumor growth was significantly inhibited. Moreover, combination treatment with an anti-PD-1 antibody and FF-10101 exhibited a far stronger antitumor effect than either treatment alone. In human cancer specimens, FF-10101 treatment reduced PD-L1 expression on TAMs, as observed in animal models. Thus, FF-10101 acts as an immunomodulatory agent that can reduce immunosuppressive TAMs and augment tumor antigen-specific T cell responses, thereby generating an immunostimulatory TME. We propose that FF-10101 is a potential candidate for successful combination cancer immunotherapy with immune checkpoint inhibitors, such as PD-1/PD-L1 blockade. To address the mechanism(s) of the tumor regression by FF-10101, comprehensive gene expression analyses was performed. Female C57BL/6J mice at 7-week-old of age were inoculated subcutaneously with a suspension of 1 × 10^6 tumor cells at the right flank (day 0). The mice were treated with or without FF-10101, and tumor samples were collected on day 8 after tumor inoculation. The experiment were conducted with three biological replicates of each group. Total RNA were sequenced using an Illumina NovaSeq6000.