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Regulated ex vivo regional gene therapy for bone repair using an inducible caspase-9 suicide gene system

Gene Therapy volume 26, pages 230–239 (2019)Cite this article

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Abstract

In order to adapt ex vivo regional gene therapy for clinical applications in orthopaedic surgery, safety issues must be considered. In this study we developed a suicide approach using a dual gene expression two step transcriptional amplification lentiviral vector (LV-TSTA) encoding BMP-2 and an inducible caspase 9 (iC9) system that selectively induces apoptosis upon activation with a chemical inducer of dimerization (CID). Transduction of rat bone marrow stromal cells (RBMSCs) with LV-TSTA-iC9/BMP-2 led to abundant BMP-2 production (90.3 ± 7.9 ng/24 h/106 cells) in vitro and stimulated bone formation in a mouse muscle pouch in the absence of CID. Moreover it was shown that CID could be used to selectively induce apoptosis in iC9-transduced cells both in vitro and in vivo. Double exposure to serial dilutions of CID decreased in vitro production of BMP-2 by 85–87% and Luc activity by 97–99% in iC9/BMP-2 or iC9/Luc-transduced cells respectively. Early administration of CID (Days 0–1 post-op) in mice implanted with iC9/BMP-2-transduced RBMSCs was effective in blocking bone formation, indicating that CID was toxic to the transduced cells. In iC9/Luc-implanted mice, late administration of two doses of CID (Days 27–28 post-op) significantly reduced the luciferase signal. The current study provides proof of concept for the potential clinical application of regulated gene therapy to promote bone repair.

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Acknowledgements

This work was supported by a National Institutes of Health grant to JRL [R01AR057076]. The authors would like to thank Ryan Park and Ivetta Vorobyova of the USC Molecular Imaging Center for their invaluable contribution in microCT and in vivo bioluminescent imaging. We would also like to thank Amy Tang for the histology sections and staining. Finally, special thanks to Dr. David Spencer for his guidance on the iC9/CID system.

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA

    Sofia Bougioukli, Venus Vakhshori, Brandon Ortega, Osamu Sugiyama & Jay Lieberman

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  1. Sofia Bougioukli
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  2. Venus Vakhshori
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  3. Brandon Ortega
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  4. Osamu Sugiyama
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  5. Jay Lieberman
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Contributions

SB participated in project development, and conducted the animal surgeries, plain radiographs, histomorphometry, data analysis and interpretation, and manuscript preparation. VV participated in project development, assisted in animal surgeries and imaging, data interpretation, and manuscript editing. BO participated in project development, generated the vectors, performed the in vitro experiments and edited the final manuscript. OS designed and generated the vectors, performed the transductions and in-vitro experiments, and edited the final manuscript. JRL designed and supervised the study, interpreted the results and edited the manuscript.

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Correspondence to Jay Lieberman.

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Conflict of interest

JRL has received royalties and has served as a paid consultant for Depuy, is a shareholder in Hip Innovation Technologies, Inc and has received royalties, financial or material support from Elsevier. The remaining authors declare that they have no conflict of interest.

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Bougioukli, S., Vakhshori, V., Ortega, B. et al. Regulated ex vivo regional gene therapy for bone repair using an inducible caspase-9 suicide gene system. Gene Ther 26, 230–239 (2019). https://doi.org/10.1038/s41434-019-0069-4

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