Figure 1 from In Situ Proinflammatory Effects of Dazostinag Alone or with Chemotherapy on the Tumor Microenvironment of Patients with Head and Neck Squamous Cell Carcinoma

Dazostinag induces a type 1 IFN response detected by (A and B) GeoMx digital spatial profiling and (C) by ISH. The 104 genes in the dazostinag signature (A) were analyzed for enrichment in biological pathways using the Molecular Signatures Database (Broad Institute, Cambridge, MA; RRID: SCR_016863). The annotation below the heatmap shows very high enrichment in IFN pathways: HALLMARK_INTERFERON_GAMMA_RESPONSE fourfold enrichment, P = 2.61E⁻⁹¹; HALLMARK_INTERFERON_ALPHA_RESPONSE threefold enrichment, P = 2.66E⁻⁶⁵; REACTOME_INTERFERON_SIGNALING sevenfold enrichment, P = 7.96E⁻⁵²; REACTOME_INTERFERON_ALPHA_BETA_SIGNALING threefold enrichment, P = 3.17E⁻⁴⁹; REACTOME_INTERFERON_GAMMA_SIGNALING fivefold enrichment, P = 1.24E⁻³⁰; WP_TYPE_II_INTERFERON_SIGNALING twofold enrichment, P = 2.69E⁻³⁰. A and B, Spatial profiling of >1,800 gene targets was performed at sites of microdose exposure for a 24-hour sample in one patient. Both pan-CK–masked areas (shown as purple and gray areas of larger squares on B) and unmasked areas (shown as smaller gray squares on B) were collected, and response heatmaps were analyzed by mask. Treatment effect was reported as the fold change over the vehicle control injection site. Genes positively associated with dazostinag exposure are strongly correlated with an induction of IFN signaling and are shown in red; those enriched in the downregulated gene set are shown in blue. C, Representative images of drug injection sites showing enrichment of IFN genes, including Cxcl10 (red) and Isg15 (yellow), after 24 hours of dazostinag monotherapy, CPPT + PTX, and dazostinag + CPPT + PTX exposure (left) and 20× magnification of the CPPT + PTX injection site (right). Cell nuclei are stained with DAPI (blue), and CIVO GLO is shown in green. CPPT, carboplatin; PTX, paclitaxel; ROI, region of interest.