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Lethal toxicity caused by expression of shRNA in the mouse striatum: implications for therapeutic design

Gene Therapy volume 18, pages 666–673 (2011)Cite this article

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Abstract

Therapeutic RNA interference (RNAi) has emerged as a promising approach for the treatment of many incurable diseases, including cancer, infectious disease or neurodegenerative disorders. Demonstration of efficacy and safety in animal models is necessary before planning human application. Our group and others have previously shown the potential of this approach for the dominantly inherited neurological disease DYT1 dystonia by achieving potent short-hairpin RNA (shRNA)-mediated silencing of the disease protein, torsinA, in cultured cells. To establish the feasibility of this approach in vivo, we pursued viral delivery of shRNA in two different mouse models. Surprisingly, intrastriatal injections of adeno-associated virus serotype 2/1 (AAV2/1) vectors expressing different shRNAs, whether targeting torsinA expression or mismatched controls, resulted in significant toxicity with progressive weight loss, motor dysfunction and animal demise. Histological analysis showed shRNA-induced neurodegeneration. Toxicity was not observed in animals that received control AAV2/1 encoding no shRNA, and was independent of genotype, occurring in both DYT1 and wild-type animals. Interestingly, the different genetic background of both mouse models influenced toxicity, being earlier and more severe in 129/SvEv than in C57BL/6 mice. In conclusion, our studies demonstrate that expression of shRNA in the mammalian brain can lead to lethal toxicity. Furthermore, the genetic background of rodents modifies their sensitivity to this form of toxicity, a factor that should be taken into consideration in the design of preclinical therapeutic RNAi trials.

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Acknowledgements

We thank Drs Beverly Davidson, Kevin Glenn, Anton McCaffrey and Henry Paulson, and members of their laboratories for their help and suggestions, Nicole Bode for her assistance with cresyl violet staining and Dr Bridgett Zimmerman for aid with statistical analyses. This work was supported by NIH/NINDS (P01NS050210 and R21NS047432), Dystonia Medical Research Foundation and Tyler's Hope For A Dystonia Cure.

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Authors and Affiliations

  1. Graduate Program in Genetics, University of Iowa, Iowa City, IA, USA

    J N Martin & P Gonzalez-Alegre

  2. Department of Neurology, University of Iowa, Iowa City, IA, USA

    N Wolken & P Gonzalez-Alegre

  3. Department of Surgery, Massachusetts General Hospital, Boston, MA, USA

    T Brown

  4. Departments of Neurology and Cell and Development Biology, University of Michigan, Ann Arbor, MI, USA

    W T Dauer

  5. Department of Neurology, Mt Sinai School of Medicine, New York, NY, USA

    M E Ehrlich

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  1. J N Martin
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Correspondence to P Gonzalez-Alegre.

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Competing interests

Janine N Martin, Nicolle Wolken and Timothy Brown declare no conflict of interest. Dr Pedro Gonzalez-Alegre's work has been funded by the NIH, Dystonia Medical Research Foundation and Tyler's Hope for a Dystonia Cure. He co-owns a patent on the technology described in this report. Dr William T Dauer's work has been funded by the NIH and Dystonia Medical Research Foundation. Dr Michelle E Ehrlich's work has been funded by the NIH.

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Martin, J., Wolken, N., Brown, T. et al. Lethal toxicity caused by expression of shRNA in the mouse striatum: implications for therapeutic design. Gene Ther 18, 666–673 (2011). https://doi.org/10.1038/gt.2011.10

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