Generation of a spontaneous murine HPV + oral cancer model with site-specific oncogene insertion using CRISPR-SONIC

Julia Tao; Jason Murray; Hsin-Fang Tu; Darrell Fan; Ya-Chea Tsai; Ming-Hung Hu; Annie A Wu; Deyin Xing; Chien-Fu Hung; T-C Wu

doi:10.1186/s13578-025-01427-5

Generation of a spontaneous murine HPV + oral cancer model with site-specific oncogene insertion using CRISPR-SONIC

Cell Biosci. 2025 Jun 18;15(1):84. doi: 10.1186/s13578-025-01427-5.

Authors

Julia Tao¹, Jason Murray¹, Hsin-Fang Tu^{1

2}, Darrell Fan¹, Ya-Chea Tsai¹, Ming-Hung Hu^{1

3}, Annie A Wu¹, Deyin Xing¹, Chien-Fu Hung^{4

5

6}, T-C Wu^{7

8

9

10

11}

Affiliations

¹ Department of Pathology, Johns Hopkins University School of Medicine, 1550 Orleans Street, CRB II 307, Baltimore, MD, 21287, USA.
² Department of Surgery, Yale School of Medicine, Yale University, New Haven, CT, USA.
³ Division of Hematology and Oncology, Department of Internal Medicine, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan.
⁴ Department of Pathology, Johns Hopkins University School of Medicine, 1550 Orleans Street, CRB II 307, Baltimore, MD, 21287, USA. chung2@jhmi.edu.
⁵ Department of Oncology, Johns Hopkins School of Medicine, Baltimore, MD, USA. chung2@jhmi.edu.
⁶ Department of Obstetrics and Gynecology, Johns Hopkins School of Medicine, Baltimore, MD, USA. chung2@jhmi.edu.
⁷ Department of Pathology, Johns Hopkins University School of Medicine, 1550 Orleans Street, CRB II 307, Baltimore, MD, 21287, USA. wutc@jhmi.edu.
⁸ Department of Oncology, Johns Hopkins School of Medicine, Baltimore, MD, USA. wutc@jhmi.edu.
⁹ Department of Obstetrics and Gynecology, Johns Hopkins School of Medicine, Baltimore, MD, USA. wutc@jhmi.edu.
¹⁰ Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins School of Medicine, Baltimore, MD, USA. wutc@jhmi.edu.
¹¹ The Johns Hopkins University School of Medicine, 1550 Orleans St, CRB II Room 309, Baltimore, MD, 21287, USA. wutc@jhmi.edu.

Abstract

Human papillomavirus associated head and neck cancer (HPV + HNC) is rising globally, emphasizing the need for improved therapeutic and screening strategies. To test novel therapies and study HPV-related disease progression, it is vital to develop relevant preclinical models. However, many fail to address critical concerns, including generating a representative immune microenvironment and adequately modeling HPV-driven malignant transformation. Recent multi-omics studies reveal the significance of HPV integration location in HPV-related carcinogenesis and highlight the necessity of targeted treatment methods. Thus, we have developed a murine model of HPV16 + HNC modifying the published CRISPR-based Somatic Oncogene kNock-In for Cancer Modeling (CRISPR-SONIC) system for precise integration of HPV oncogenes. We showed that CRISPR-SONIC knock-in of Kras^G12D, HPV16 E6 and E7, and a luciferase reporter at the murine β-actin 3'-UTR locus could induce spontaneous buccal tumors with sarcomatous morphology under transient or selective immunosuppression. Both preventative and therapeutic pNGVL4a-CRT/E7(detox) DNA vaccination could induce HPV16 E7-specific immune response and reduce tumor growth. Furthermore, CRISPR-SONIC knock-in of HPV16 E6 and E7 with co-delivery of HNC-relevant oncogenes AKT and c-Myc produced tumors in NSG mice capturing the characteristic carcinomic morphology of HPV + HNC. Overall, our model offers a robust platform for evaluating new therapies and exploring HPV-related carcinogenesis.

Keywords: Buccal tumor; CRISPR-SONIC; E6; E7; HPV16; Head and neck cancer; Human papillomavirus; Immunotherapy; Murine model; Ras.

Abstract

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