Integration of oncogenes via sleeping beauty as a mouse model of HPV16þ oral tumors and immunologic control

Yi Hsin Lin, Ming Chieh Yang, Ssu Hsueh Tseng, Rosie Jiang, Andrew Yang, Emily Farmer, Shiwen Peng, Talia Henkle, Yung Nien Chang, Chien-Fu Hung, Tzyy Choou Wu

Research output: Contribution to journalArticle

Abstract

Human papillomavirus type 16 (HPV16) is the etiologic factor for cervical cancer and a subset of oropharyngeal cancers. Although several prophylactic HPV vaccines are available, no effective therapeutic strategies to control active HPV diseases exist. Tumor implantation models are traditionally used to study HPV-associated buccal tumors. However, they fail to address precancerous phases of disease progression and display tumor microenvironments distinct from those observed in patients. Previously, K14-E6/E7 transgenic mouse models have been used to generate spontaneous tumors. However, the rate of tumor formation is inconsistent, and the host often develops immune tolerance to the viral oncoproteins. We developed a preclinical, spontaneous, HPV16þ buccal tumor model using submucosal injection of oncogenic plasmids expressing HPV16-E6/E7, NRasG12V, luciferase, and sleeping beauty (SB) transposase, followed by electroporation in the buccal mucosa. We evaluated responses to immunization with a pNGVL4a-CRT/E7(detox) therapeutic HPV DNA vaccine and tumor cell migration to distant locations. Mice transfected with plasmids encoding HPV16-E6/E7, NRasG12V, luciferase, and SB transposase developed tumors within 3 weeks. We also found transient anti-CD3 administration is required to generate tumors in immunocompetent mice. Bioluminescence signals from luciferase correlated strongly with tumor growth, and tumors expressed HPV16-associated markers. We showed that pNGVL4a-CRT/E7(detox) administration resulted in antitumor immunity in tumor-bearing mice. Lastly, we demonstrated that the generated tumor could migrate to tumor-draining lymph nodes. Our model provides an efficient method to induce spontaneous HPVþ tumor formation, which can be used to identify effective therapeutic interventions, analyze tumor migration, and conduct tumor biology research.

Original languageEnglish (US)
Pages (from-to)305-319
Number of pages15
JournalCancer immunology research
Volume6
Issue number3
DOIs
StatePublished - Mar 1 2018

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Beauty
Human papillomavirus 16
Oncogenes
Neoplasms
Luciferases
Transposases
Papillomavirus Vaccines
Cheek
Plasmids
Oropharyngeal Neoplasms
Immune Tolerance
DNA Vaccines
Tumor Microenvironment
Electroporation
Oncogene Proteins
Mouth Mucosa

ASJC Scopus subject areas

  • Immunology
  • Cancer Research

Cite this

Integration of oncogenes via sleeping beauty as a mouse model of HPV16þ oral tumors and immunologic control. / Lin, Yi Hsin; Yang, Ming Chieh; Tseng, Ssu Hsueh; Jiang, Rosie; Yang, Andrew; Farmer, Emily; Peng, Shiwen; Henkle, Talia; Chang, Yung Nien; Hung, Chien-Fu; Wu, Tzyy Choou.

In: Cancer immunology research, Vol. 6, No. 3, 01.03.2018, p. 305-319.

Research output: Contribution to journalArticle

Lin, Yi Hsin ; Yang, Ming Chieh ; Tseng, Ssu Hsueh ; Jiang, Rosie ; Yang, Andrew ; Farmer, Emily ; Peng, Shiwen ; Henkle, Talia ; Chang, Yung Nien ; Hung, Chien-Fu ; Wu, Tzyy Choou. / Integration of oncogenes via sleeping beauty as a mouse model of HPV16þ oral tumors and immunologic control. In: Cancer immunology research. 2018 ; Vol. 6, No. 3. pp. 305-319.
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abstract = "Human papillomavirus type 16 (HPV16) is the etiologic factor for cervical cancer and a subset of oropharyngeal cancers. Although several prophylactic HPV vaccines are available, no effective therapeutic strategies to control active HPV diseases exist. Tumor implantation models are traditionally used to study HPV-associated buccal tumors. However, they fail to address precancerous phases of disease progression and display tumor microenvironments distinct from those observed in patients. Previously, K14-E6/E7 transgenic mouse models have been used to generate spontaneous tumors. However, the rate of tumor formation is inconsistent, and the host often develops immune tolerance to the viral oncoproteins. We developed a preclinical, spontaneous, HPV16{\th} buccal tumor model using submucosal injection of oncogenic plasmids expressing HPV16-E6/E7, NRasG12V, luciferase, and sleeping beauty (SB) transposase, followed by electroporation in the buccal mucosa. We evaluated responses to immunization with a pNGVL4a-CRT/E7(detox) therapeutic HPV DNA vaccine and tumor cell migration to distant locations. Mice transfected with plasmids encoding HPV16-E6/E7, NRasG12V, luciferase, and SB transposase developed tumors within 3 weeks. We also found transient anti-CD3 administration is required to generate tumors in immunocompetent mice. Bioluminescence signals from luciferase correlated strongly with tumor growth, and tumors expressed HPV16-associated markers. We showed that pNGVL4a-CRT/E7(detox) administration resulted in antitumor immunity in tumor-bearing mice. Lastly, we demonstrated that the generated tumor could migrate to tumor-draining lymph nodes. Our model provides an efficient method to induce spontaneous HPV{\th} tumor formation, which can be used to identify effective therapeutic interventions, analyze tumor migration, and conduct tumor biology research.",
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AU - Farmer, Emily

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AU - Wu, Tzyy Choou

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