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Synergistic antitumor effect on cervical cancer by rational combination of PD1 blockade and CRISPR-Cas9-mediated HPV knockout

Cancer Gene Therapy volume 27, pages 168–178 (2020)Cite this article

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Abstract

Targeted therapy results in objective responses in cervical cancer. However, the responses are short. In contrast, treatment with immune checkpoint inhibitors results in a lower responses rate, but the responses tend to be more durable. Based on these findings, we hypothesized that HPV16 E6/E7-targeted therapy may synergize with the PD-1 pathway blockade to enhance antitumor activity. To test hypothesis, we described for the first time the effects of the CRISPR/Cas9 that was targeted to the HPV and PD1 in vitro and in vivo. Our data showed that gRNA/cas9 targeted HPV16 E6/E7 induced cervical cancer cell SiHa apoptosis, and suggested that overexpression of PD-L1, induced by HPV16 E6/E7, may be responsible for lymphocyte dysfunction. In established SiHa cell- xenografted humanized SCID mice, Administration of gRNA-PD-1 together with gRNA-HPV16 E6/E7 treatment improved the survival and suppressed the tumor growth obviously. In addition, combination treatment increased the population of dendritic cells, CD8+ and CD4+ T lymphocyte cells. According, it enhanced the expression of Th1-associated immune-stimulating genes while reducing the transcription of regulatory/suppressive immune genes, reshaping tumor microenvironment from an immunosuppressive to a stimulatory state. These results demonstrate potent synergistic effects of combination therapy using HPV16 E6/E7-targeted therapy and immune checkpoint blockade PD1, supporting a direct translation of this combination strategy in clinic for the treatment of cervical cancer.

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Acknowledgements

This study was supported by grants from the National Natural Science Foundation of China (Grant nos. 81602295 to Shuai Zhen). This study was supported by grants from Key Research and Development Program of Shaanxi Province of China (2017ZDXM‐SF‐24‐1).

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  1. These authors contributed equally: Shuai Zhen, Jiaojiao Lu

Authors and Affiliations

  1. Center for Translational Medicine, Key Laboratory for Tumor Precision Medicine of Shaanxi Province, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China

    Shuai Zhen, Jiaojiao Lu & Xu Li

  2. Center of Medical Genetics, Northwest Women and Children’s Hospital, Xi’an, China

    Shuai Zhen, Jiaojiao Lu & Xu Li

  3. Department of Pharmacology and Toxicology, Beijing Institute of Radiation Medicine, Beijing, China

    Yun-Hui Liu

  4. Center of Laboratory Medicine, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China

    Wei Chen

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  1. Shuai Zhen
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Correspondence to Xu Li.

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This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The study was consistent with the principles used to relieve the pains of animals, as well as using the least number of animals as possible.

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Zhen, S., Lu, J., Liu, YH. et al. Synergistic antitumor effect on cervical cancer by rational combination of PD1 blockade and CRISPR-Cas9-mediated HPV knockout. Cancer Gene Ther 27, 168–178 (2020). https://doi.org/10.1038/s41417-019-0131-9

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