sting agonist sr717  (TargetMol)

Journal: Cell Reports Medicine

Article Title: Interferon-driven CAF reprogramming augments immunogenic response to neoadjuvant radiotherapy in colorectal cancer

doi: 10.1016/j.xcrm.2025.102251

Figure Lengend Snippet: ilCAFs mediate radiation-induced senescence via the IFN-γ/STAT1 pathway (A) Representative histogram plots (left) and quantification (right) showing mean fluorescence intensity (MFI) of IRF1 or CCL4 in CAFs isolated from treatment-naive patients with rectal cancer, analyzed by flow cytometry. Comparisons include vehicle control, RT, IFN-γ, IFN-γ + RT, STAT1 inhibitor (STAT1i), and RT + STAT1i, all evaluated at 48 h post-RT ( n = 3 per group). (B) Representative immunofluorescent staining images (left) and quantification (right) depicting the expression of α-SMA, IRF1, CCL4, or STAT1 in primary CAFs treated with vehicle, IFN-γ, IFN-γ ± RT, STAT1i, or STAT1i ± RT at specified time points ( n = 5 per group). Scale bars, 10 μm. (C) Western blot images showing the expression levels of cGAS, p-STAT1/STAT1, p-STING/STING, IRF1, and CCL4 in primary CAFs treated with IFN-γ (20 ng/mL) or STAT1i (5 μM) combined with RT for 48 h. (D) GSEA depicting the upregulated STING pathway signature in ilCAFs compared to other CAFs from rectal cancers. (E) Heatmap depicts a list of differentially expressed genes sourced from inflammatory modulation, type II IFN, and TNF-α between vehicle- and RT-treated groups. (F) Representative flow cytometric dot plots displaying the MFI of IRF1 or CCL4 in CAFs isolated from treatment-naive rectal cancer patients ( n = 3 per group), comparing vehicle control, RT, STING agonist SR717, and SR717 + RT at 48 h post-RT treatment. (G) Western blot images depicting the expression levels of cGAS, p-STAT1/STAT1, p-STING/STING, and IRF1 in primary CAFs treated with SR717 (3 μM). (H) Bubble plots illustrating upregulated pathways in fibroblasts co-cultured with tumor cell-derived media (TCMs) after RT + SR717 treatment compared to RT alone, based on GO biological processes. Pathways were filtered using adjusted p values (P adj < 0.05). (I) Quantification (right) of flow cytometry analysis of CD8 + T cells, GZMB + CD8 + T cells, CD11c + DCs, and CD103 + CD11c + cDC1s in T cells or DCs co-cultured with mouse fibroblasts from the indicated combination treatment groups ( n = 3 per group). For (A), (B), (F), and (I), one-way ANOVA was performed. Data are presented as mean ± SEM and are representative of at least three independent experiments. A p value less than 0.05 indicates statistical significance. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Similarly, for the therapeutic combination of radiation with the STING agonist SR717 (TargetMol, USA), SR717 was intraperitoneally injected at a dose of 30 mg/kg every other day after radiation.

Techniques: Fluorescence, Isolation, Flow Cytometry, Control, Staining, Expressing, Western Blot, Cell Culture, Derivative Assay

Journal: Cell Reports Medicine

Article Title: Interferon-driven CAF reprogramming augments immunogenic response to neoadjuvant radiotherapy in colorectal cancer

doi: 10.1016/j.xcrm.2025.102251

Figure Lengend Snippet: STING knockout in CAFs modulates the stromal landscape and enhances T cell infiltration (A and B) Histogram plots (left) and quantification of MFI of IRF1 (A) or CCL4 (B) in primary fibroblasts isolated from treatment-naive wild-type (WT) and Tmem173 −/− (STING knockout) mice, analyzed by flow cytometry. Comparisons include vehicle, RT, STING agonist (SR717), and combination therapy (SR717 + RT) in Tmem173 −/− fibroblasts and vehicle and RT in WT fibroblasts post-RT (vehicle group in A: n = 4; all other groups: n = 3 per group). (C–G) Impact of RT on the growth of established MC38 colorectal tumors co-inoculated with Tmem173 −/− or WT fibroblasts in syngeneic C57BL/6 mice. (C) Schematic of the experimental design. (D) Average tumor growth curve. (E) Tumor weight at endpoint. (F) Tumor-draining lymph node sizes. (G) Modified Kaplan-Meier survival curves for each treatment group ( n = 5 per group). (H) Percentage of CD4 + T cells, CD8 + T cells, and cDC1s among CD45 + tumor-infiltrating cells at day 14 post-RT treatment ( n = 5 per group). (I) Representative multiplex immunofluorescence images (left) and quantification (right) of IRF1 + CAFs (green: COL1A, red: IRF1) and CCL4 + CAFs (green: COL1A, red: CCL4). Scale bars, 100 μm ( n = 5 per group). For (A), (B), (E), (F), (H), and (I), one-way ANOVA was performed. For (D), a two-way ANOVA was conducted. For (G), the log rank test was performed. Data are presented as mean ± SEM. Results are representative of at least three independent experiments. A p value less than 0.05 indicates statistical significance. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Similarly, for the therapeutic combination of radiation with the STING agonist SR717 (TargetMol, USA), SR717 was intraperitoneally injected at a dose of 30 mg/kg every other day after radiation.

Techniques: Knock-Out, Isolation, Flow Cytometry, Modification, Multiplex Assay, Immunofluorescence

Journal: Cell Reports Medicine

Article Title: Interferon-driven CAF reprogramming augments immunogenic response to neoadjuvant radiotherapy in colorectal cancer

doi: 10.1016/j.xcrm.2025.102251

Figure Lengend Snippet: STING agonist augments the presence of ilCAFs and enhances endogenous anti-tumor immunity to synergize with radiotherapy for enhanced efficacy (A) Experimental design illustrating the treatment regimen administered to C57BL/6 mice bearing MC38 colorectal tumors. (B) Average tumor growth curve in syngeneic mice co-inoculated with fibroblasts and treated with vehicle, SR717 (STING agonist), RT, or the combination SR717 with RT. (C) Tumor weights at endpoint. (D) Modified Kaplan-Meier survival curves representing outcomes for each treatment group ( n = 5 per group). (E) Dot plots illustrating expression signatures of different CAF clusters in the mouse colorectal tumor model, determined by bulk RNA-seq. Violin plots depict cDC1s signature expression in MC38 tumors following the indicated treatments ( n = 3 per group). (F and G) Representative multiplex IF images (F) and quantification (G) of STING + CAFs (green: STING, red: COL1A) and p-STAT1 + CAFs (green: p-STAT1, red: COL1A; n = 5 per group). Scale bars, 100 μm. (H) Heatmap depicting the expression levels of multiple signaling pathways in tumors treated with vehicle control, SR717, RT, and RT + SR717 ( n = 3 per group). (I and J) Representative multiplex IF images (left) and quantification (right) of endpoint tumors stained for colocalized CD103 (green) and DC (red) (I) and CD4 (red) and CD8 (green) (J) ( n = 5 per group). Nuclei were stained with DAPI (blue). Scale bar, 100 μm. (K) Proportion of CD4 + T cells, CD8 + T cells, and cDC1s among CD45 + cells in tumors and tumor-draining lymph nodes at day 12 post-RT in vehicle control, SR717, RT, and RT + SR717 groups ( n = 5 mice/group). For (C), (E), (G), (I), (J), and (K), one-way ANOVA was performed. For (B), a two-way ANOVA was conducted. For (D), the log rank test was utilized. Data are presented as mean ± SEM. Results are representative of at least three independent experiments. A p value less than 0.05 indicates statistical significance. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Similarly, for the therapeutic combination of radiation with the STING agonist SR717 (TargetMol, USA), SR717 was intraperitoneally injected at a dose of 30 mg/kg every other day after radiation.

Techniques: Modification, Expressing, RNA Sequencing, Multiplex Assay, Protein-Protein interactions, Control, Staining

Journal: Cell Reports Medicine

Article Title: Interferon-driven CAF reprogramming augments immunogenic response to neoadjuvant radiotherapy in colorectal cancer

doi: 10.1016/j.xcrm.2025.102251

Figure Lengend Snippet: STING agonist enhances the therapeutic efficacy of radiotherapy in spontaneous colorectal cancer mouse models (A) Experimental timeline illustrating the treatment regimen administered to Apc min/+ mice bearing spontaneous colorectal tumors. (B) Hematoxylin and eosin (H&E) staining of small intestinal sections from 17-week-old mice. Scale bars indicate 1 mm (top) and 100 μm (bottom). (C) Quantification of tumor number and tumor burden in the small intestine and total intestinal tract ( n = 4 per group), comparing treatments with vehicle control, radiotherapy (RT), STING agonist (SR717), and combination therapy (RT + SR717). (D) Schematic timeline of the experimental design evaluating the impact of combination therapy on dextran sodium sulfate (DSS)-induced tumorigenesis in C57BL/6J wild-type mice injected intraperitoneally with azoxymethane (AOM, 10 mg/kg). (E) Representative images of colorectal tumors treated with RT versus RT + SR717 ( n = 5 per group). (F) H&E staining of colorectal sections from DSS-induced tumorigenesis. Scale bars: 1 mm (top) and 100 μm (bottom). (G) Quantification of tumor burden and tumor number in the colorectal and total intestinal tract ( n = 5 per group), comparing RT with RT + SR717 treatment groups. (H and I) Representative multiplex IF images (H) and quantification (I) showing the localization of STING + CAFs and p-STAT1 + CAFs (green: STING, red: p-STAT1, orange: COL1A, blue: DAPI) ( n = 5 per group). Scale bars indicate 100 μm. Data are presented as mean ± SEM. Statistical analyses included one-way ANOVA for (C) and Student’s t tests for (G) and (I). A p value less than 0.05 indicates statistical significance. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Similarly, for the therapeutic combination of radiation with the STING agonist SR717 (TargetMol, USA), SR717 was intraperitoneally injected at a dose of 30 mg/kg every other day after radiation.

Techniques: Drug discovery, Staining, Control, Injection, Multiplex Assay

Journal: Cell Reports Medicine

Article Title: Interferon-driven CAF reprogramming augments immunogenic response to neoadjuvant radiotherapy in colorectal cancer

doi: 10.1016/j.xcrm.2025.102251

Figure Lengend Snippet: STING agonist reverses radioresistance in colorectal tumors (A) Treatment protocol: C57BL/6 mice bearing radioresistant MC38 tumors received 20 Gy of RT, followed by treatment with a STING agonist. (B–D) Average tumor growth curves (B), tumor weight at the endpoint (C), and tumor-draining lymph node sizes (D) were evaluated across different treatment cohorts in a syngeneic transplant mouse model ( n = 5 per group). (E) Heatmap illustrating the transcriptional profiles of STING and type II IFN signaling pathway-associated genes in radioresistant tumors treated with vehicle control, SR717, RT, or RT + SR717, as revealed by bulk RNA-seq ( n = 3 per group). (F) Bubble plots illustrating the upregulated pathways in tumors treated with RT + SR717 relative to RT alone, based on known biological functions in GO databases. Pathways were filtered using adjusted p values (P adj < 0.05). (G) Representative multiplex IF image (left) and quantification (right) demonstrating the localization of STING + CAFs and p-STAT1 + CAFs within the tumor microenvironment (green: STING, red: p-STAT1, orange: COL1A, blue: DAPI). Scale bar, 100 μm ( n = 5 per group). (H) Quantification of CD4 + T cells, CD8 + T cells, and cDC1s among CD45 + cells in tumors and tumor-draining lymph nodes from vehicle control, SR717, RT, and RT + SR717 treatment groups at day 14 post-RT ( n = 5 per group). Data are presented as mean ± SEM. Statistical analyses were performed using one-way ANOVA for (C), (D), (G), and (H) and two-way ANOVA for (B). A p value less than 0.05 indicates statistical significance. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Similarly, for the therapeutic combination of radiation with the STING agonist SR717 (TargetMol, USA), SR717 was intraperitoneally injected at a dose of 30 mg/kg every other day after radiation.

Techniques: Control, RNA Sequencing, Multiplex Assay