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MicroRNA-150-regulated vectors allow lymphocyte-sparing transgene expression in hematopoietic gene therapy

Gene Therapy volume 19, pages 915–924 (2012)Cite this article

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Abstract

Endogenous microRNA (miRNA) expression can be exploited for cell type-specific transgene expression as the addition of miRNA target sequences to transgenic cDNA allows for transgene downregulation specifically in cells expressing the respective miRNAs. Here, we have investigated the potential of miRNA-150 target sequences to specifically suppress gene expression in lymphocytes and thereby prevent transgene-induced lymphotoxicity. Abundance of miRNA-150 expression specifically in differentiated B and T cells was confirmed by quantitative reverse transcriptase PCR. Mono- and bicistronic lentiviral vectors were used to investigate the effect of miRNA-150 target sequences on transgene expression in the lymphohematopoietic system. After in vitro studies demonstrated effective downregulation of transgene expression in murine B220+ B and CD3+ T cells, the concept was further verified in a murine transplant model. Again, marked suppression of transgene activity was observed in B220+ B and CD4+ or CD8+ T cells whereas expression in CD11b+ myeloid cells, lin− and lin−/Sca1+ progenitors, or lin−/Sca1+/c-kit+ stem cells remained almost unaffected. No toxicity of miRNA-150 targeting in transduced lymphohematopoietic cells was noted. Thus, our results demonstrate the suitability of miRNA-150 targeting to specifically suppress transgene expression in lymphocytes and further support the concept of miRNA targeting for cell type-specific transgene expression in gene therapy approaches.

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Acknowledgements

We thank Matthias Ballmaier and his team from the Core-Facility Cell-Sorting of Hannover Medical School for cell sorting and Doreen Lüttge (Hannover) for excellent technical assistance. This work was supported by grants from the Deutsche Forschungsgemeinschaft: Cluster of Excellence REBIRTH (Exc 62/1), SPP1230 Grant MO 886/3-1 (UM and TM).

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Authors and Affiliations

  1. REBIRTH Cluster-of-Excellence, Research Group Reprogramming, Hannover Medical School, Hannover, Germany

    N Lachmann, J Jagielska, S Brennig, N Pfaff & T Moritz

  2. Department of Experimental Hematology, REBIRTH Cluster-of-Excellence, Hannover Medical School, Hannover, Germany

    D Heckl, T Maetzig, U Modlich & A Schambach

  3. REBIRTH Cluster-of-Excellence, JRG Stem Cell Biology, Hannover Medical School, Hannover, Germany

    T Cantz

  4. Max Planck Institute for Molecular Biomedicine, Münster, Germany

    T Cantz

  5. Division of Regenerative Medicine, San Raffaele Telethon Institute for Gene Therapy, Stem Cells and Gene Therapy, San Raffaele Scientific Institute, Milan, Italy

    B Gentner

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  1. N Lachmann
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Correspondence to T Moritz.

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Lachmann, N., Jagielska, J., Heckl, D. et al. MicroRNA-150-regulated vectors allow lymphocyte-sparing transgene expression in hematopoietic gene therapy. Gene Ther 19, 915–924 (2012). https://doi.org/10.1038/gt.2011.148

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  • DOI: https://doi.org/10.1038/gt.2011.148

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