Tropism, intracerebral distribution, and transduction efficiency of HIV- and SIV-based lentiviral vectors after injection into the mouse brain: a qualitative and quantitative in vivo study

Juraj Hlavatý,Zbyněk Tonar,Matthias Schweizer,Sylvia Panitz,Björn-Philipp Kloke,Silke Schüle,Helga Petznek,Rudolf Moldzio,Matthias Renner,Kirsti Witter

doi:10.1007/s00418-017-1569-1

Abstract

Lentiviruses are suitable to transfer potential therapeutic genes into non-replicating cells such as neurons, but systematic in vivo studies on transduction of neural cells within the complete brain are missing. We analysed the distribution of transduced cells with respect to brain structure, virus tropism, numbers of transduced neurons per brain, and influence of the Vpx or Vpr accessory proteins after injection of vectors based on SIVsmmPBj, HIV-2, and HIV-1 lentiviruses into the right striatum of the mouse brain. Transduced cells were found ipsilaterally around the injection canal, in corpus striatum and along corpus callosum, irrespective of the vector type. All vectors except HIV-2SEW transduced also single cells in the olfactory bulb, hippocampus, and cerebellum. Vector HIV-2SEW was the most neuron specific. However, vectors PBjSEW and HIV-1SEW transduced more neurons per brain (means 41,299 and 32,309) than HIV-2SEW (16,102). In the presence of Vpx/Vpr proteins, HIV-2SEW(Vpx) and HIV-1SEW(Vpr) showed higher overall transduction efficiencies (30,696 and 27,947 neurons per brain) than PBjSEW(Vpx) (6636). The distances of transduced cells from the injection canal did not differ among the viruses but correlated positively with the numbers of transduced neurons. The presence of Vpx/Vpr did not increase the numbers of transduced neurons. Parental virus type and the vector equipment seem to influence cellular tropism and transduction efficiency. Thus, precision of injection and choice of virus pseudotype are not sufficient when targeted lentiviral vector transduction of a defined brain cell population is required.

Highlights

Many neurodegenerative diseases such as Alzheimer’s, Parkinson’s, and Huntington’s disease still can only be treated inadequately
The transduction potential of all viruses has been confirmed for both neurons and glial cells by positive double immunofluorescence for enhanced green fluorescent protein gene (EGFP) and neuronal nuclear antigen (NeuN) and EGFP and glial fibrillary acidic protein (GFAP), respectively (Online Resource 3)
The transduction efficiency for neurons ranged between 20% for PBjSEW(Vpx) and 37% for PBjSEW and HIV2SEW (Online Resource 4)

Summary

Introduction

Many neurodegenerative diseases such as Alzheimer’s, Parkinson’s, and Huntington’s disease still can only be treated inadequately. Gene therapy could hold the promise of therapeutic progress (Blömer et al 1996; Wong et al 2006; Eleftheriadou and Mazarakis 2015). As many neurodegenerative diseases develop with progressive, incurable degeneration of neuronal cells, gene therapy approaches for the treatment of such disorders require vector systems that can efficiently transduce non-dividing neurons. Lentiviral vectors have been shown to do so and by integrating their genome into the genome of the transduced cells they supposedly provide permanent expression of the transferred therapeutic gene in these cells and in the progeny of transduced dividing neuronal precursors Lentiviral vectors have been shown to do so and by integrating their genome into the genome of the transduced cells they supposedly provide permanent expression of the transferred therapeutic gene in these cells and in the progeny of transduced dividing neuronal precursors (Lundberg et al. Vol.:(0123456789)

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