Journal: Research Square

Article Title: Stepwise Enhancement of HPV16 E6/E7 mRNA Vaccine Efficacy Using HSV-1 gD Epitope Incorporation and Immune-Modulating Agents

doi: 10.21203/rs.3.rs-9890379/v1

Figure Lengend Snippet: (A) Experimental design. All mice received two doses of HSV-1 gD mRNA administered one week apart. Mice were then assigned to receive PBS, mRNA-UB-E6/E7-gD, ADU-S100, or ADU-S100 + mRNA-UB-E6/E7-gD. One week after the final vaccination, mice were challenged subcutaneously with TC-1 cells. ADU-S100 (10 μg/dose) was administered intratumorally beginning 7 days after tumor implantation, when tumors reached approximately 5 × 5 mm, for a total of three treatments. (B) Tumor volumes of individual mice at the experimental endpoint. (C) Kaplan–Meier survival analysis of the indicated treatment groups. (D) Cytotoxic T-lymphocyte (CTL) activity against TC-1 target cells measured 24 h after assay initiation. Effector-to-target (E) ratios are indicated. (E) IFN-γ production in splenocyte culture supernatants measured by ELISA 24 h after assay initiation. E ratios are indicated. (F, G) Flow cytometric analysis of HPV16 E7-specific CD8 + T cells in tumor-infiltrating lymphocytes (F) and splenocytes (G), determined by HPV16 E7 tetramer staining. (H) Quantification of CD4 + and CD8 + tumor-infiltrating cells by immunohistochemistry. Positive cells were quantified using ImageJ from randomly selected microscopic fields and expressed as the number of positive cells per field. Statistical significance: ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: ADU-S100 [2′3′-c-di-AM(PS)2 (Rp,Rp); MedChemExpress; cat. no. HY-12885] was administered intratumorally at 10 μg per mouse for three total doses given every 3 days.

Techniques: Tumor Implantation, Activity Assay, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: The cGAS–STING pathway in cancer immunotherapy: prognostic value and therapeutic potential

doi: 10.3389/fimmu.2026.1760947

Figure Lengend Snippet: Overview of cGAS–STING signaling activation in tumor cells and the classes of STING-targeting therapeutics. STING pathway activation links cytosolic DNA sensing to the induction of type I interferons and pro-inflammatory cytokines, driving innate and adaptive antitumor immunity. Exosomes, mtDNA, micronuclei, and other DAMPs activate cGAS to produce cGAMP, which engages STING and triggers ER-to-Golgi signaling. Pharmacologic STING agonists—including CDN analogs, non-CDN small molecules, and ADC-based payloads—target distinct points along this cascade to amplify immune activation and tumor rejection. (i) Cyclic dinucleotide (CDN) STING agonists: CDN analogs (e.g., ADU-S100, 2′3′-cGAMP analogues, MK-1454, GSK-532, and SB-11285) mimic natural cGAMP and directly engage the STING ligand-binding domain on the ER to initiate downstream signaling. (ii) Non-CDN small-molecule STING agonists: non-CDN agonists (e.g., MSA-1, MSA-2, GSK-3745417, MK-2118, SR-717, TAK-676, and XNS-281) stabilize the active conformation of STING—often independent of cGAS—and exhibit improved cellular penetration relative to CDNs. (iii) Antibody–drug conjugate (ADC) STING agonists: ADC-based STING agonists (e.g., CRD-5500, TAK-500, and XMT-2056) deliver STING-activating payloads selectively to tumor or immune cells through tumor-specific surface antigens, enabling targeted intratumoral STING activation while minimizing systemic toxicity. (iv) Integrated antitumor effects: Collectively, these therapeutic classes converge on the cGAS–STING axis to enhance tumor-intrinsic signaling, innate immune activation, and cytotoxic T cell-mediated tumor rejection. IRF3, interferon regulatory factor-3; GTP, guanosine triphosphate; cGAS, cyclic GMP-AMP synthase; ATP, adenosine triphosphate; STING, stimulator of interferon genes; cGAMP, cyclin guanosine monophosphate–adenosine monophosphate; DAMPs, danger-associated molecular patterns; NF-κB, nuclear factor kappa-light chain-enhancer of activated B cells; mtDNA, mitochondrial DNA, inhibitor of kappa B; CDNs, cyclic dinucleotides; TBK, TANK-binding kinase.

Article Snippet: One of the most extensively studied is ADU-S100 (MIW815), a synthetic CDN developed by Novartis (formerly Aduro Biotech) ( ).

Techniques: Activation Assay, Analogues, Ligand Binding Assay, Binding Assay

Journal: Molecular Oncology

Article Title: Stimulator of interferon genes agonist augmented antitumor immunity of osimertinib in Egfr ‐mutated lung cancer

doi: 10.1002/1878-0261.70264

Figure Lengend Snippet: Antitumor effect of ADU‐S100 in Egfr ‐mutant lung cancer mouse model. (A) Representative images of NK1.1 and CD8 immunohistochemistry (IHC) staining on tumors from Egfr ‐mutant mice treated with phosphate‐buffered saline (PBS) (100 μL, intratumoral administration [i.t.], Day 1 as vehicle) or ADU‐S100 (50 μg, i.t., Day 1) after 4 days. The NK1.1 + and CD8 + cells were quantified using the imagej software. Error bars represent the standard error ( n = 5 fields of view per group). Scale bars: 100 μm. ** P < 0.01, Student's t ‐test. Data shown are representative of three independent experiments with similar results. (B) The effect of ADU‐S100. Tumor growth in the Egfr ‐mutant lung cancer mouse model treated with PBS (100 μL, i.t., Day 1 as vehicle) or ADU‐S100 (50 μg, i.t., Day 1) ( n = 3 tumors per group, 3 mice per group). Error bars represent the standard error. * P < 0.05, Student's t ‐test. Data shown are representative of three independent experiments with similar results. (C) Combination effect of osimertinib and ADU‐S100. Tumor growth in the Egfr ‐mutant lung cancer model treated with PBS (100 μL, i.t., Day 1 vehicle; n = 4 tumors from 2 mice per group), osimertinib (15 mg·kg −1 ·day −1 , oral gavage [p.o.], 14 days; n = 6 tumors from 3 mice per group), ADU‐S100 (50 μg, i.t., Day 1; n = 6 tumors from 3 mice per group), or combination of osimertinib (15 mg·kg −1 ·day −1 , p.o., 14 days; n = 6 tumors from 3 mice per group) and ADU‐S100 (50 μg, i.t., Day 1). A vertical dashed line at Day 8 indicates the time point used for Bliss independence model analysis. Error bars represent the standard error. * P < 0.05, Student's t ‐test. Data shown are representative of two independent experiments with similar results. (D) Representative images of NK1.1, CD8, and CD11c IHC staining on Egfr ‐mutant lung cancer tumors from mice treated with PBS (100 μL, i.t., Day 1 as vehicle), osimertinib (15 mg·kg −1 ·day −1 , p.o.), ADU‐S100 (50 μg, i.t., Day 1), and combination of osimertinib (15 mg·kg −1 ·day −1 , p.o.) and ADU‐S100 (50 μg, i.t., Day 1) for 4 days. The NK1.1 + , CD8 + and CD11c + cells were quantified using imagej software. Error bars represent the standard error ( n = 5 fields of view per group). Scale bars: 100 μm. ns = not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, one‐way analysis of variance (ANOVA) with the post hoc Tukey test. Data shown are representative of three independent experiments with similar results. Serial sections of the same tumor tissue were used for the various immunostainings shown in this panel.

Article Snippet: ADU‐S100 disodium salt (cat. no. HY‐12885A) was purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Mutagenesis, Immunohistochemistry, Saline, Software

Journal: Molecular Oncology

Article Title: Stimulator of interferon genes agonist augmented antitumor immunity of osimertinib in Egfr ‐mutated lung cancer

doi: 10.1002/1878-0261.70264

Figure Lengend Snippet: Enhancement of antitumor immunity via CD8 + cells and NK1.1 + cells in combination therapy. (A) Tumor growth in Egfr ‐mutant lung cancer mice treated with osimertinib (15 mg·kg −1 ·day −1 , oral gavage [p.o.], 7 days/week) for 14 days and observed for 7 days with anti‐CD8 or anti‐NK1.1 antibodies. n = 6 tumors per group, 3 mice per group. Data shown are representative of two independent experiments with similar results. Error bars represent the standard error. ns = not significant, ** P < 0.01, Student's t ‐test. (B) Tumor growth in mice treated with osimertinib (15 mg·kg −1 ·day −1 , p.o., 7 days/week) for 14 days and ADU‐S100 (50 μg, intratumoral administration [i.t.]., Day 1) and observed for 7 days with anti‐CD8 or NK1.1 antibodies. n = 6 tumors per group, 3 mice per group. Data shown are representative of two independent experiments with similar results. Error bars represent the standard error. * P < 0.05, ** P < 0.01, Student's t ‐test. (C) Representative images of Granzyme B immunohistochemistry (IHC) staining treated with osimertinib (15 mg·kg −1 ·day −1 , p.o., 7 days/week) or combination of osimertinib (15 mg·kg −1 ·day −1 , p.o., 7 days/week) and ADU‐S100 (50 μg, i.t., on Day 1). Tumors were harvested on Day 4 after treatment initiation. Granzyme B + cells were quantified using the imagej software. Scale bars: 100 μm. Error bars represent the standard error ( n = 5 fields of view per group). Data shown are representative of two independent experiments with similar results. (D) Representative images of EGFR and interferon regulatory factor 3 (IRF3) IHC staining treated with osimertinib (15 mg·kg −1 ·day −1 , p.o.) or combination of osimertinib (15 mg·kg −1 ·day −1 , p.o.) and ADU‐S100 (50 μg, i.t., on Day 1) Tumors were harvested on Day 4 after treatment initiation. %Area of EGFR + and IRF3 + cells was quantified using the imagej software. Scale bars: 100 μm. Error bars represent the standard error ( n = 5 fields of view per group). Data shown are representative of two independent experiments with similar results. Serial sections of the same tumor tissue were used for the various immunostainings shown in this panel. (E) Crystal violet‐stained cells after drug loading for 4 days. Cancer cells were treated with 0.2% dimethyl sulfoxide, 1 μmol·L −1 osimertinib, and/or 10 μmol·L −1 ADU‐S100.

Article Snippet: ADU‐S100 disodium salt (cat. no. HY‐12885A) was purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Mutagenesis, Immunohistochemistry, Software, Staining

Journal: Molecular Oncology

Article Title: Stimulator of interferon genes agonist augmented antitumor immunity of osimertinib in Egfr ‐mutated lung cancer

doi: 10.1002/1878-0261.70264

Figure Lengend Snippet: Abscopal effects induced by osimertinib and local injection of ADU‐S100 in Egfr ‐mutant lung cancer mouse model. (A) Schematic image of the transplantation and treatment schedule of the Egfr ‐mutant lung cancer mouse model. 1st = first, 2nd = secondary. (B) Secondary tumor growth n = 3 tumors per group, 3 mice per group. Data shown are representative of two independent experiments with similar results. Error bars represent the standard error. ns = not statistically significant, Student's t ‐test. (C) Representative images of NK1.1 and CD8 immunohistochemistry (IHC) staining on secondary tumors, 7 days after transplantation. The NK1.1 + and CD8 + cells were quantified using imagej software. Error bars represent the standard error ( n = 5 fields of view per group). Scale bars: 100 μm. Data shown are representative of two independent experiments with similar results. Serial sections of the same tumor tissue were used for the various immunostainings shown in this panel. (D) Schematic image of the transplantation and treatment schedule of the Egfr ‐mutant lung cancer mouse model with osimertinib combination. 1st = first, 2nd = secondary. (E) Secondary tumor growth n = 4 tumors per group, 4 mice per group. Data shown are representative of three independent experiments with similar results. Error bars represent the standard error. ** P < 0.01, Student's t ‐test. (F) Representative images of NK1.1 and CD8 IHC staining on the secondary tumors, 7 days after transplantation. The NK1.1 + and CD8 + cells were quantified using the imagej software. Error bars represent the standard error ( n = 7 fields of view per group). Scale bars: 100 μm. Data shown are representative of two independent experiments with similar results. Serial sections of the same tumor tissue were used for the various immunostainings shown in this panel.

Article Snippet: ADU‐S100 disodium salt (cat. no. HY‐12885A) was purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Injection, Mutagenesis, Transplantation Assay, Immunohistochemistry, Software

Journal: Molecular Oncology

Article Title: Stimulator of interferon genes agonist augmented antitumor immunity of osimertinib in Egfr ‐mutated lung cancer

doi: 10.1002/1878-0261.70264

Figure Lengend Snippet: Abscopal effect induced by osimertinib and ADU‐S100 combination therapy is mediated by CD8 + cells. (A) Flow cytometry analysis of NK1.1 + and CD8 + cells isolated from secondary tumors, 7 days after transplantation. For NK1.1 + cells, n = 8 per group; for CD8 + cells, n = 12 per group. Error bars represent the standard error. ns = not significant, * P < 0.05, Student's t ‐test. The NK1.1 + cell data are the integrated results of two independent experiments with matching conditions. The CD8 + cell data are the integrated results of three independent experiments with matching conditions. (B) Representative images of Granzyme B immunohistochemistry staining on the secondary tumors, 7 days after transplantation. The Granzyme B + cells were quantified using the imagej software. Error bars represent the standard error ( n = 5 fields of view per group). Scale bars: 100 μm. Data shown are representative of two independent experiments with similar results. (C) Secondary tumor growth with anti‐CD8 or anti‐NK1.1 antibodies. For NK1.1 + cells and CD8 + cells, n = 8 tumors from 8 mice per group; for Control, n = 6 tumors from six mice per group. The data are the integrated results of two independent experiments with matching conditions. Error bars represent the standard error. ns = not significant, *** P < 0.001, Student's t ‐test.

Article Snippet: ADU‐S100 disodium salt (cat. no. HY‐12885A) was purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Flow Cytometry, Isolation, Transplantation Assay, Immunohistochemistry, Staining, Software, Control