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Analysis of neuronal proliferation, migration and differentiation in the postnatal brain using equine infectious anemia virus-based lentiviral vectors

Gene Therapy volume 16, pages 1021–1033 (2009)Cite this article

Abstract

Ongoing neurogenesis in discrete sectors of the adult central nervous system depends on the mitotic activity of an elusive population of adult stem cells. The existence of adult neural stem cells provides an alternative approach to transplantation of embryonic stem cells in cell-based therapies. Owing to the limited intrinsic fate of adult stem cells and inhibitory nature of the adult brain for neurogenesis, accommodation for circuit replacement in the brain will require genetic and epigenetic manipulation. Here, we show that a replication-incompetent Equine Infectious Anemia Virus (EIAV) is highly suitable for stable and persistent gene transfer to adult neural stem cells. The transduced regions were free of long-lasting neuroimmune responses to EIAV. Transduction in the subventricular zone was specific to the stem cell niche, but spared the progeny of adult neural stem cells that includes transit amplifying progenitors (TAPs) and migrating neuroblasts. With time, EIAV-transduced stem cells passed on the transgene to TAPs and migrating neuroblasts, which ultimately differentiated into neurons in the olfactory bulbs. We show that EIAV is highly suitable for discovery and assessment of mechanisms that regulate proliferation, migration and differentiation in the postnatal brain.

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Acknowledgements

We thank Dr David Eisenstat (University of Manitoba) for providing the DLX2 antibody. We are grateful to Dr Roman Giger (University of Michigan) for providing the Sema3A construct. Drs Rick Meeker (UNC-Chapel Hill) and Lola Hudson (North Carolina State University) provided invaluable antibodies and discussion on neuroimmune and neuroinflammatory responses to EIAV. Natalie Surzenko helped with the NRG2 analysis. We also thank Dr Chris McGahan for critical reading of the manuscript. JCO acknowledges research support from the National Institutes of Health HL 51818 and sponsored research funds from the Oxford BioMedica Ltd. The initial stages of this work was supported by National Institute of Health Grant MH060929 to ESA. Most of this work was supported by NCSU start up funds to TG.

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  1. B V Jacquet and M Patel: These authors contributed equally.

Authors and Affiliations

  1. Department of Molecular Biomedical Sciences, North Carolina State University, Raleigh, NC, USA

    B V Jacquet, M Iyengar, H Liang, B Therit, R Salinas-Mondragon & H T Ghashghaei

  2. Department of Medicine, University of North Carolina, Chapel Hill, NC, USA

    M Patel & J C Olsen

  3. Gill Center for Biomolecular Science, Indiana University, Bloomington, IN, USA

    C Lai

  4. Neuroscience Research Center, University of North Carolina, Chapel Hill, NC, USA

    E S Anton

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  1. B V Jacquet
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Correspondence to H T Ghashghaei.

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Supplementary Information accompanies the paper on Gene Therapy website (http://www.nature.com/gt)

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Jacquet, B., Patel, M., Iyengar, M. et al. Analysis of neuronal proliferation, migration and differentiation in the postnatal brain using equine infectious anemia virus-based lentiviral vectors. Gene Ther 16, 1021–1033 (2009). https://doi.org/10.1038/gt.2009.58

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