Article Figures & Data
Figures
- Figure 1.
Inhibition of colony-stimulating factor-1 receptor reduces M2 macrophage polarization. Bone marrow derived monocytes were cultured with granulocyte-macrophage colony-stimulating factor, followed by M2 induction with interleukin-4 and -13 with or without BLZ-945. Representative fluorescence-activated sorting plots are shown. Cells were initially gated by the pan-macrophage markers CD45 and F/480 to identify total macrophages. The subset of macrophages that stained for CD11b and CD206 were then selected to identify M2 macrophages. Experiments were performed in triplicates.
- Figure 2.
Inhibition of colony-stimulating factor-1 receptor potentiates the impact of radiotherapy (RT) on tumor volume. TRP murine glioblastoma cells were stereotactically injected into the right hemisphere of 6- to 8-week-old mice. On day 14, treatment with vehicle, BLZ-945 (200 mg/kg), RT (6 Gy × 3 fractions), or BLZ-945 + RT was initiated, followed by daily treatment with vehicle or BLZ-945 for 2 weeks. A: Representative hematoxylin and eosin (H&E) images from each treatment group. Pink-stained tissue shows normal brain. Blue-stained tissue shows tumor cells. B: Tumor burden expressed as a percentage of normal brain volume based on H&E staining. Brains were harvested on day 10 post-treatment initiation and sectioned every 10 µM through the tumor and adjacent brain tissue. Slides were imaged and segmented. An iterative learning algorithm was employed to identify normal brain tissue and tumor cells. p-Values for significant differences are shown.
- Figure 3.
Inhibition of colony-stimulating factor-1 receptor potentiates radiotherapy (RT) and improves survival. TRP murine glioblastoma cells were stereotactically injected into the right hemisphere of 6- to 8-week-old mice. On day 14, treatment with vehicle, BLZ-945 (200 mg/kg), RT (6 Gy × 3 fractions), or BLZ-945 with RT was initiated, followed by daily treatment with vehicle or BLZ-945 for 3 weeks. Mice were sacrificed if they became symptomatic, lost ≥20% weight, or reached 14 weeks. B: Sample magnetic resonance imaging of tumors prior to, and 2 weeks post-treatment (Tx). Tumors were contoured on the T1 sequence and volumes were estimated by the number of voxels. C: Boxplot of tumor volumes in each treatment group before treatment initiation. D: Boxplot of tumor volumes in each treatment group 2 weeks after initiation of treatment. p-Values for significant differences are shown.
- Figure 4.
Kaplan–Meier estimates of survival for each treatment arm. Dashed lines denote median overall survival.
- Figure 5.
Immunophenotyping of the tumor microenvironment. From the survival experiment, brains were harvested at the time of sacrifice, tumors were extracted, and fluorescence-activated sorting was used to analyze the presence of various immune cells in each treatment arm. A: Macrophages, identified as CD45-, CDF4/80-, and CD11b-positive cells. B: M2 macrophages, identified by the markers in (A) in addition to CD206. C: CD8+ T-cells, identified as CD3- and CD8-positive cells. D: M2 to CD8 cell ratio. E: Regulatory T-cells (Tregs), identified as CD4- and CD25-positive cells. p-Values for significant differences are shown.
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