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Inhibition of Gli2 suppresses tumorigenicity in glioblastoma stem cells derived from a de novo murine brain cancer model

Cancer Gene Therapy volume 28, pages 1339–1352 (2021)Cite this article

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Abstract

The prognosis of glioblastoma remains poor despite intensive research efforts. Glioblastoma stem cells (GSCs) contribute to tumorigenesis, invasive capacity, and therapy resistance. Leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5), a stem cell marker, is involved in the maintenance of GSCs, although the properties of Lgr5-positive GSCs remain unclear. Here, the Sleeping-Beauty transposon-induced glioblastoma model was used in Lgr5-GFP knock-in mice identify GFP-positive cells in neurosphere cultures from mouse glioblastoma tissues. Global gene expression analysis showed that Gli2 was highly expressed in GFP-positive GSCs. Gli2 knockdown using lentiviral-mediated shRNA downregulated Hedgehog-related and Wnt signaling pathway-related genes, including Lgr5; suppressed tumor cell proliferation and invasion capacity; and induced apoptosis. Pharmacological Gli inhibition with GANT61 suppressed tumor cell proliferation. Silencing Gli2 suppressed the tumorigenicity of GSCs in an orthotopic transplantation model in vivo. These findings suggest that Gli2 affects the Hedgehog and Wnt pathways and plays an important role in GSC maintenance, suggesting Gli2 as a therapeutic target for glioblastoma treatment.

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Fig. 1: Lgr5-GFP-positive glioblastoma stem cells exhibit higher tumorigenicity.
Fig. 2: Activation of the Hh signaling pathway affects Lgr5 expression.
Fig. 3: Gli2 knockdown affects the Hh and Wnt pathways.
Fig. 4: Gli2 knockdown suppresses the growth of GSCs.
Fig. 5: GANT61 suppresses proliferation and induces apoptosis.
Fig. 6: Gli2 knockdown suppresses tumorigenicity in vivo.

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Acknowledgements

The authors are grateful to Satomi Toma, Hitoshi Iwasaki, Shinsaku Nakamura, Koichi Uesaka, and Noriko Saito for experimental support.

Funding

This work was supported by the Japan Society for the Promotion of Science, grant numbers 20K07623 and 16K08722; the Ministry of Education, Culture, Sports, Science and Technology–Supported Program for the Strategic Research Foundation at Private Universities 2015–2019; and the Kyoto Pharmaceutical University Fund for the Promotion of Collaborative Research.

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Authors and Affiliations

  1. Department of Clinical Oncology, Kyoto Pharmaceutical University, Kyoto, Japan

    Seisuke Tanigawa, Chiami Moyama, Shota Ando, Hiromi Ii & Susumu Nakata

  2. Department of Neurosurgery, Kyoto Prefectural University Graduate School of Medical Science, Kyoto, Japan

    Seisuke Tanigawa, Hayato Takeuchi, Takumi Yamanaka, Yoshinobu Takahashi & Naoya Hashimoto

  3. Department of Microbiology, Faculty of Medicine, Kindai University, Osaka, Japan

    Mitsugu Fujita

  4. Division of Pathophysiology, Aichi Cancer Center Research Institute, Nagoya, Japan

    Yasushi Kojima, Teruaki Fujishita & Masahiro Aoki

  5. Department of Cancer Physiology, Nagoya University Graduate School of Medicine, Nagoya, Japan

    Masahiro Aoki

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Tanigawa, S., Fujita, M., Moyama, C. et al. Inhibition of Gli2 suppresses tumorigenicity in glioblastoma stem cells derived from a de novo murine brain cancer model. Cancer Gene Ther 28, 1339–1352 (2021). https://doi.org/10.1038/s41417-020-00282-5

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