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Cellular and Molecular Biology

Novel strategy to target glioblastoma-initiating cells using a braintropic adeno-associated virus carrying a miR-dependent genome-editing system

British Journal of Cancer volume 132, pages 1100–1109 (2025)Cite this article

Subjects

Abstract

Background

Since glioblastoma (GBM)-initiating cells (GICs) were identified as the cells-of-origin for GBM, various GIC factors have been analyzed as potential therapeutic targets. However, these targets are also present in normal cells outside of the brain, raising concerns about potential side effects when directly targeted. The aim of this study is to develop a novel method that specifically eradicates GICs with reducing side effects.

Methods

We selected micoRNAs (miRs) that are significantly decreased in GICs compared to normal cells and developed a genome-editing (GE) system that knocks out a functional GIC factor in a miR-dependent manner (miR-dependent GE). Additionally, we developed mosaic-capsids that consist of braintropic and universal capsids, which deliver genes into GIC brain tumors.

Results

Systemic administration of the mosaic-capsids Adeno-associated virus (AAV) carrying a miR-dependent GFP expression cassette selectively expressed GFP in GICs transplanted into the brains of immunodeficient mice, without expression in either mouse brain cells or non-brain tissues. The mosaic-capsids AAV carrying a miR-dependent GE prevented GIC tumorigenesis in the brain and extended the survival time of tumor-bearing mice.

Conclusion

These data indicate that the mosaic-capsids AAV containing a miR-dependent GE represents a novel therapeutic virus for GBM with fewer side effects.

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Fig. 1: Identification of miR-219a-2-3p and miR-340-5p as candidate miRs for the miR-dependent gene expression system.
Fig. 2: GE AAV encoding miRCS induces cell-cycle arrest and cell death in GICs.
Fig. 3: miR-dependent AAV-GFP selectively express GFP in GIC brain tumor cells in NOG mice.
Fig. 4: Anti-tumorigenic activity of miR-dependent GE AAV with mosaic capsids in GIC brain tumors.

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Data availability

Additional data supporting the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank Dr. Feng Zhang for providing pX601-AAV-CMV NLS-SaCas9-NLS-3xHA-bGHpA;U6 BsaI-sgRNA through Addgene, and Dr. Toshio Kitamura for pMY-IRES-EGFP vector.

Funding

This work was partly supported by AMED Practical Research for Innovative Cancer Control (17ck0106236h0002), JSPS KAKENHI Grant-in-Aid for Scientific Research (20H03559), and the Joint Research Program of the Institute for Genetic Medicine Hokkaido University (all to TK).

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Authors and Affiliations

  1. Division of Stem Cell Biology, Institute for Genetic Medicine, Hokkaido University, Sapporo, Hokkaido, 060-0815, Japan

    Zhe Wang, Peilin Zou, Zhenzhong Chen, Jiahui Hou, You Lee Son & Toru Kondo

  2. Department of Neurosurgery, Ehime University Graduate School of Medicine, Toon, Ehime, 791-0295, Japan

    Daisuke Yamashita

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  1. Zhe Wang
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Contributions

TK conceived the study and designed the experiments. ZW, PZ, ZC, JH, YLS, DY and TK performed the experiments and analyzed data. ZC, JH, YLS and TK have contributed to the interpretation of the results. TK wrote the manuscript. All authors have read and approved the final version.

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Correspondence to Toru Kondo.

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The authors declare no competing interests.

Ethics approval and consent to participate

Five primary human glioma samples, including three GBMs, one anaplastic astrocytoma (AA), and one diffuse oligodendroglioma (DO), were collected from Ehime University Hospital. Informed consent was obtained from all patients according to the Research Ethics Committee guidelines (approval number: 1208009) and the specimens were used to prepare glioma lines, GICs (E3, E6, E16), AA, and DO. The glioma lines were used in compliance with the research guidelines of the Ehime University Graduate School of Medicine and the Institute for Genetic Medicine of Hokkaido University.

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Wang, Z., Zou, P., Chen, Z. et al. Novel strategy to target glioblastoma-initiating cells using a braintropic adeno-associated virus carrying a miR-dependent genome-editing system. Br J Cancer 132, 1100–1109 (2025). https://doi.org/10.1038/s41416-025-03007-3

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