Combination of Vaccine With IL-12-Armed Oncolytic Virus SKV-012 Synergistically Potentiates Immune Responses in HPV-Associated Malignancies

Abstract

Currently, patients with advanced-stage or refractory human papillomavirus (HPV)-associated malignancies have few therapeutic options. Despite therapeutic HPV vaccines having been investigated, the lack of appreciable efficacy highlights the urgent need to develop more effective strategies. Here, we developed an immuno-oncotherapy for HPV-induced tumors based on an adenoviral (Ad)-vectored therapeutic vaccine that contains concatemeric T cell epitopes, and evaluated oncolytic viruses (OV) as potential approach to enhance vaccine efficacy. We observed that the therapeutic vaccine encoding the HPV E7 oncoprotein epitope (Ad-E7P) significantly inhibited tumor growth in HPV-induced murine models by inducing systemic antitumor CD8+T cell responses and promoting the formation of tertiary lymphoid structures in peritumoral regions. We then evaluated the potential of combining the vaccine with an interleukin-12 (IL-12)–armed oncolytic herpesvirus (SKV-012) in preclinical models. The combination therapy elicited potent antitumor responses by inducing antigen-specific T-cell expansion, remodeling the tumor microenvironment, and generating immune memory, which led to tumor clearance. Overall, these findings support that the vaccine synergizes with the OV as an effective approach to enhance antitumor immunity in HPV-associated malignancies.

Keywords

antitumor responses / HPV-associated malignancies / oncolytic viruses / therapeutic vaccines / tumor microenvironment

References

[1] A. A. Shamseddine, B. Burman, N. Y. Lee, D. Zamarin, and N. Riaz, “Tumor Immunity and Immunotherapy for HPV-Related Cancers,” Cancer Discovery 11, no. 8 (2021): 1896–1912.

[2] F. Wei, D. Georges, I. Man, I. Baussano, and G. M. Clifford, “Causal Attribution of human Papillomavirus Genotypes to Invasive Cervical Cancer Worldwide: A Systematic Analysis of the Global Literature,” Lancet (London, England) 404, no. 10451 (2024): 435–444.

[3] R. L. Ferris and W. Westra, “Oropharyngeal Carcinoma With a Special Focus on HPV-Related Squamous Cell Carcinoma,” Annual Review of Pathology 18 (2023): 515–535.

[4] M. Falcaro, A. Castañon, B. Ndlela, et al., “The Effects of the National HPV Vaccination Programme in England, UK, on Cervical Cancer and Grade 3 Cervical Intraepithelial Neoplasia Incidence: A Register-based Observational Study,” Lancet (London, England) 398, no. 10316 (2021): 2084–2092.

[5] M. Saraiya, E. R. Unger, T. D. Thompson, et al., “US Assessment of HPV Types in Cancers: Implications for Current and 9-valent HPV Vaccines,” Journal of the National Cancer Institute 107, no. 6 (2015): djv086.

[6] C. L. Trimble, M. P. Morrow, K. A. Kraynyak, et al., “Safety, Efficacy, and Immunogenicity of VGX-3100, a Therapeutic Synthetic DNA Vaccine Targeting human Papillomavirus 16 and 18 E6 and E7 Proteins for Cervical Intraepithelial Neoplasia 2/3: A Randomised, Double-blind, Placebo-controlled Phase 2b Trial,” Lancet (London, England) 386, no. 10008 (2015): 2078–2088.

[7] M. L. Bagarazzi, J. Yan, M. P. Morrow, et al., “Immunotherapy Against HPV16/18 Generates Potent TH1 and Cytotoxic Cellular Immune Responses,” Science Translational Medicine 4, no. 155 (2012): 155ra138.

[8] J. Sadoff, G. Gray, A. Vandebosch, et al., “Final Analysis of Efficacy and Safety of Single-Dose Ad26.COV2.S,” The New England Journal of Medicine 386, no. 9 (2022): 847–860.

[9] L. Coughlan, E. J. Kremer, and D. M. Shayakhmetov, “Adenovirus-based Vaccines-a Platform for Pandemic Preparedness Against Emerging Viral Pathogens,” Molecular Therapy: The Journal of the American Society of Gene Therapy 30, no. 5 (2022): 1822–1849.

[10] D. van Riel and E. de Wit, “Next-generation Vaccine Platforms for COVID-19,” Nature Materials 19, no. 8 (2020): 810–812.

[11] E. Borcoman, A. Lalanne, J. P. Delord, et al., “Phase Ib/II Trial of Tipapkinogene Sovacivec, a Therapeutic human papillomavirus16-vaccine, in Combination With avelumab in Patients With Advanced human papillomavirus16-positive Cancers,” European Journal of Cancer (Oxford, England: 1990) 191 (2023): 112981.

[12] T. C. van der Sluis, F. J. van Haften, S. van Duikeren, et al., “Delayed Vaccine-induced CD8(+) T Cell Expansion by Topoisomerase I Inhibition Mediates Enhanced CD70-dependent Tumor Eradication,” Journal for Immunotherapy of Cancer 11, no. 11 (2023): e007158.

[13] M. Weller, N. Butowski, D. D. Tran, et al., “Rindopepimut With Temozolomide for Patients With Newly Diagnosed, EGFRvIII-expressing Glioblastoma (ACT IV): A Randomised, Double-blind, International Phase 3 Trial,” The Lancet Oncology 18, no. 10 (2017): 1373–1385.

[14] E. A. Mittendorf, B. Lu, M. Melisko, et al., “Efficacy and Safety Analysis of Nelipepimut-S Vaccine to Prevent Breast Cancer Recurrence: A Randomized, Multicenter, Phase III Clinical Trial,” Clinical Cancer Research: An Official Journal of the American Association for Cancer Research 25, no. 14 (2019): 4248–4254.

[15] R. Ramlau, E. Quoix, J. Rolski, et al., “A Phase II Study of Tg4010 (Mva-Muc1-Il2) in Association With Chemotherapy in Patients With Stage III/IV Non-small Cell Lung Cancer,” Journal of Thoracic Oncology: Official Publication of the International Association for the Study of Lung Cancer 3, no. 7 (2008): 735–744.

[16] S. Meulewaeter, I. Aernout, J. Deprez, et al., “Alpha-galactosylceramide Improves the Potency of mRNA LNP Vaccines Against Cancer and Intracellular Bacteria,” Journal of Controlled Release: Official Journal of the Controlled Release Society 370 (2024): 379–391.

[17] S. Kreiter, M. Diken, A. Selmi, et al., “FLT3 ligand Enhances the Cancer Therapeutic Potency of Naked RNA Vaccines,” Cancer Research 71, no. 19 (2011): 6132–6142.

[18] R. Fu, R. Qi, H. Xiong, et al., “Combination Therapy With Oncolytic Virus and T Cells or mRNA Vaccine Amplifies Antitumor Effects,” Signal Transduction and Targeted Therapy 9, no. 1 (2024): 118.

[19] K. DePeaux and G. M. Delgoffe, “Integrating Innate and Adaptive Immunity in Oncolytic Virus Therapy,” Trends in Cancer 10, no. 2 (2024): 135–146.

[20] H. L. Kaufman, F. J. Kohlhapp, and A. Zloza, “Oncolytic Viruses: A New Class of Immunotherapy Drugs,” Nature Reviews Drug Discovery 14, no. 9 (2015): 642–662.

[21] R. Li, P. H. Shah, T. F. Stewart, et al., “Oncolytic Adenoviral Therapy plus Pembrolizumab in BCG-unresponsive Non-muscle-invasive Bladder Cancer: The Phase 2 CORE-001 Trial,” Nature Medicine 30, no. 8 (2024): 2216–2223.

[22] K. Harrington, D. J. Freeman, B. Kelly, J. Harper, and J. C. Soria, “Optimizing Oncolytic Virotherapy in Cancer Treatment,” Nature Reviews Drug Discovery 18, no. 9 (2019): 689–706.

[23] M. Liu, S. Hu, N. Yan, K. D. Popowski, and K. Cheng, “Inhalable Extracellular Vesicle Delivery of IL-12 mRNA to Treat Lung Cancer and Promote Systemic Immunity,” Nature Nanotechnology 19, no. 4 (2024): 565–575.

[24] B. Brook, V. Duval, S. Barman, et al., “Adjuvantation of a SARS-CoV-2 mRNA Vaccine With Controlled Tissue-specific Expression of an mRNA Encoding IL-12p70,” Science Translational Medicine 16, no. 757 (2024): eadm8451.

[25] G. Trinchieri, “Interleukin-12 and the Regulation of Innate Resistance and Adaptive Immunity,” Nature Reviews Immunology 3, no. 2 (2003): 133–146.

[26] S. Bevers, S. A. A. Kooijmans, E. Van de Velde, et al., “mRNA-LNP Vaccines Tuned for Systemic Immunization Induce Strong Antitumor Immunity by Engaging Splenic Immune Cells,” Molecular Therapy 30, no. 9 (2022): 3078–3094.

[27] Z. Jiang, N. Yang, and J. Jin, “Preclinical and Clinical Evaluation of Intratumoral Injection of an IL-12 Expressing SKV-012 Oncolytic Virus for Advanced Solid Tumors,” Journal for Immunotherapy of Cancer 13, no. 6 (2025): e011642.

[28] L. M. Draper, M. L. Kwong, A. Gros, et al., “Targeting of HPV-16+ Epithelial Cancer Cells by TCR Gene Engineered T Cells Directed Against E6,” Clinical Cancer Research: An Official Journal of the American Association for Cancer Research 21, no. 19 (2015): 4431–4439.

[29] N. B. Nagarsheth, S. M. Norberg, A. L. Sinkoe, et al., “TCR-engineered T Cells Targeting E7 for Patients With Metastatic HPV-associated Epithelial Cancers,” Nature Medicine 27, no. 3 (2021): 419–425.

[30] M. Hiasa, M. Abe, A. Nakano, et al., “GM-CSF and IL-4 Induce Dendritic Cell Differentiation and Disrupt Osteoclastogenesis Through M-CSF Receptor Shedding by Up-regulation of TNF-alpha Converting Enzyme (TACE),” Blood 114, no. 20 (2009): 4517–4526.

[31] J. S. W. Borgers, D. Lenkala, V. Kohler, et al., “Personalized, Autologous Neoantigen-specific T Cell Therapy in Metastatic Melanoma: A Phase 1 Trial,” Nature Medicine 31, no. 3 (2025): 881–893.

[32] D. A. Braun, G. Moranzoni, V. Chea, et al., “A Neoantigen Vaccine Generates Antitumour Immunity in Renal Cell Carcinoma,” Nature 639, no. 8054 (2025): 474–482.

[33] P. D. Katsikis, K. J. Ishii, and C. Schliehe, “Challenges in Developing Personalized Neoantigen Cancer Vaccines,” Nature Reviews Immunology 24, no. 3 (2024): 213–227.

[34] M. J. Welters, T. C. van der Sluis, H. van Meir, et al., “Vaccination During Myeloid Cell Depletion by Cancer Chemotherapy Fosters Robust T Cell Responses,” Science Translational Medicine 8, no. 334 (2016): 334ra52.

[35] L. Maldonado, J. E. Teague, M. P. Morrow, et al., “Intramuscular Therapeutic Vaccination Targeting HPV16 Induces T Cell Responses That Localize in Mucosal Lesions,” Science Translational Medicine 6, no. 221 (2014): 221ra13.

[36] M. J. Atherton, K. B. Stephenson, J. K. Nikota, et al., “Preclinical Development of Peptide Vaccination Combined With Oncolytic MG1-E6E7 for HPV-associated Cancer,” Vaccine 36, no. 16 (2018): 2181–2192.

[37] C. S. Eberhardt, H. T. Kissick, M. R. Patel, et al., “Functional HPV-specific PD-1(+) Stem-Like CD8 T Cells in Head and Neck Cancer,” Nature 597, no. 7875 (2021): 279–284.

[38] S. M. Norberg and C. S. Hinrichs, “Engineered T Cell Therapy for Viral and Non-viral Epithelial Cancers,” Cancer Cell 41, no. 1 (2023): 58–69.

[39] S. Lanini, S. Capone, A. Antinori, et al., “GRAd-COV2, a Gorilla adenovirus-based Candidate Vaccine Against COVID-19, Is Safe and Immunogenic in Younger and Older Adults,” Science Translational Medicine 14, no. 627 (2022): eabj1996.

[40] C. Dold, L. Marsay, N. Wang, et al., “An Adenoviral-vectored Vaccine Confers Seroprotection Against Capsular Group B Meningococcal Disease,” Science Translational Medicine 15, no. 701 (2023): eade3901.

[41] E. J. Kremer, “Pros and Cons of Adenovirus-Based SARS-CoV-2 Vaccines,” Molecular Therapy: The Journal of the American Society of Gene Therapy 28, no. 11 (2020): 2303–2304.

[42] S. Capone, F. M. Fusco, S. Milleri, et al., “GRAd-COV2 Vaccine Provides Potent and Durable Humoral and Cellular Immunity to SARS-CoV-2 in Randomized Placebo-controlled Phase 2 Trial,” Cell Reports Medicine 4, no. 6 (2023): 101084.

[43] M. Lechner, J. Liu, L. Masterson, and T. R. Fenton, “HPV-associated Oropharyngeal Cancer: Epidemiology, Molecular Biology and Clinical Management,” Nature Reviews Clinical Oncology 19, no. 5 (2022): 306–327.

[44] A. T. Ruffin, H. Li, L. Vujanovic, D. P. Zandberg, R. L. Ferris, and T. C. Bruno, “Improving Head and Neck Cancer Therapies by Immunomodulation of the Tumour Microenvironment,” Nature Reviews Cancer 23, no. 3 (2023): 173–188.

[45] J. W. Youn, S. Y. Hur, J. W. Woo, et al., “Pembrolizumab plus GX-188E Therapeutic DNA Vaccine in Patients With HPV-16-positive or HPV-18-positive Advanced Cervical Cancer: Interim Results of a Single-arm, Phase 2 Trial,” Lancet Oncology 21, no. 12 (2020): 1653–1660.

[46] K. Ye, F. Li, R. Wang, et al., “An Armed Oncolytic Virus Enhances the Efficacy of Tumor-infiltrating Lymphocyte Therapy by Converting Tumors to Artificial Antigen-presenting Cells in Situ,” Molecular Therapy: The Journal of the American Society of Gene Therapy 30, no. 12 (2022): 3658–3676.

[47] P. K. Bommareddy, H. Wakimoto, R. L. Martuza, H. L. Kaufman, S. D. Rabkin, and D. Saha, “Oncolytic herpes Simplex Virus Expressing IL-2 Controls Glioblastoma Growth and Improves Survival,” Journal for Immunotherapy of Cancer 12, no. 4 (2024): e008880.

[48] D. A. Vignali, L. W. Collison, and C. J. Workman, “How Regulatory T Cells Work,” Nature Reviews Immunology 8, no. 7 (2008): 523–532.

[49] A. Mantovani, P. Allavena, F. Marchesi, and C. Garlanda, “Macrophages as Tools and Targets in Cancer Therapy,” Nature Reviews Drug Discovery 21, no. 11 (2022): 799–820.

[50] G. N. Medina, T. de Los Santos, and F. Díaz-San Segundo, “Generation of Replication Deficient Human Adenovirus 5 (Ad5) Vectored FMD Vaccines,” Methods in Molecular Biology (Clifton, NJ) 2465 (2022): 155–175.

[51] K. Y. Lin, F. G. Guarnieri, and K. F. Staveley-O'Carroll, “Treatment of Established Tumors With a Novel Vaccine That Enhances Major Histocompatibility Class II Presentation of Tumor Antigen,” Cancer Research 56, no. 1 (1996): 21–26.

[52] Z. Zhang, N. Yang, H. Lu, et al., “Improved Antitumor Effects Elicited by an Oncolytic HSV-1 Expressing a Novel B7H3nb/CD3 BsAb,” Cancer Letters 588 (2024): 216760.

RIGHTS & PERMISSIONS

2026 The Author(s). MedComm published by Sichuan International Medical Exchange & Promotion Association (SCIMEA) and John Wiley & Sons Australia, Ltd.