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One-year follow-up of transgene expression by integrase-defective lentiviral vectors and their therapeutic potential in spinocerebellar ataxia model mice

Gene Therapy volume 21, pages 820–827 (2014)Cite this article

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Abstract

We examined integrase-defective lentiviral vectors (IDLVs) with a mutant (D64V) integrase in terms of their residual integration capability, the levels and duration of transgene expression and their therapeutic potential in comparison to wild-type lentiviral vectors (WTLVs) with a wild-type integrase gene. Compared with WTLVs, the IDLV-mediated proviral integration into host-cell chromosomes was approximately 1/3850 in HeLa cells and approximately 1/111 in mouse cerebellar neurons in vivo. At 2 months, transgene expression by IDLVs in the mouse cerebellum was comparable to that by WTLVs, but then significantly decreased. The mRNA levels at 6 and 12 months after injection in IDLV-infected cerebella were approximately 26% and 5%, respectively, of the mRNA levels in WTLV-injected cerebella. To examine the therapeutic potential, IDLVs or WTLVs expressing a molecule that enhances the ubiquitin-proteasome pathway were injected into the cerebella of spinocerebellar ataxia type 3 model mice (SCA3 mice). IDLV-injected SCA3 mice showed a significantly improved rotarod performance even at 1 year after-injection. Immunohistochemistry at 1 year after injection showed a drastic reduction of mutant aggregates in Purkinje cellsfrom IDLV-injected, as well as WTLV-injected, SCA3 mice. Our results suggest that because of the substantially reduced risk of insertional mutagenesis, IDLVs are safer and potentially effective as gene therapy vectors.

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Acknowledgements

We thank Junko Sugiyama, Asako Onishi and Hiromi Hirai for maintaining and genotyping the mutant mice. This work was supported by a grant from the Funding Program for Next Generation World-Leading Researchers (LS021) (to HH) and a Grant-in-Aid from the Japan Society for the Promotion of Science (JSPS); Grant-in-Aid for Young Scientists (B) KAKENHI 22700374).

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  1. H Saida, Y Matsuzaki and K Takayama: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Neurophysiology, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan

    H Saida, Y Matsuzaki, K Takayama, A Iizuka, A Konno & H Hirai

  2. Laboratory of Molecular Biochemistry, Graduate School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan

    S Yanagi

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  1. H Saida
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Correspondence to H Hirai.

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Saida, H., Matsuzaki, Y., Takayama, K. et al. One-year follow-up of transgene expression by integrase-defective lentiviral vectors and their therapeutic potential in spinocerebellar ataxia model mice. Gene Ther 21, 820–827 (2014). https://doi.org/10.1038/gt.2014.60

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