Targeted transgene expression in rat brain using lentiviral vectors

J Neurosci Res. 2003 Sep 15;73(6):876-85. doi: 10.1002/jnr.10719.

Abstract

Direct gene transfer to the adult brain is dependent on vectors that transduce non-dividing cells, such as lentiviral vectors. Another aspect of the development of gene therapy to the brain is the need for cell-specific transgene expression. Expression from vesicular stomatitis virus G-protein (VSV-G) pseudotyped lentiviral vectors has been reported to be mainly neuron specific in the brain. We constructed cell-specific lentiviral vectors using the neuron-specific enolase (rNSE) or the glial fibrillary acidic protein (hGFAP) promoters and compared them to the ubiquitous human cytomegalovirus promoter (hCMV), a hybrid CMV/beta-actin promoter (CAG) and the promoter for human elongation factor 1 alpha (EF1 alpha). Our results showed that the hGFAP promoter was expressed only in glial cells, whereas rNSE was purely neuron specific, showing that VSV-G is pantropic in the rat striatum. We conclude that the VSV-G allows transduction of both glial and neuronal cells and the promoter dictates in what cell type the transgene will be expressed. The expression of transgenes exclusively in astrocytes would allow for local delivery of secreted transgene products, such as glial cell line-derived neurotrophic factor (GDNF), circumventing the anterograde transport that may induce unwanted side effects.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Animals
  • Antigens / metabolism
  • Brain / anatomy & histology
  • Brain / metabolism*
  • Brain / virology
  • Cell Count / methods
  • Cell Line / metabolism
  • Cell Line / virology
  • Female
  • Flow Cytometry / instrumentation
  • Flow Cytometry / methods
  • Gene Expression
  • Gene Transfer Techniques
  • Genetic Vectors / genetics
  • Glial Fibrillary Acidic Protein / genetics
  • Glial Fibrillary Acidic Protein / metabolism
  • Green Fluorescent Proteins
  • Humans
  • Immunohistochemistry / methods
  • Lentivirus / genetics*
  • Luminescent Proteins / metabolism
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / metabolism
  • Peptide Elongation Factor 1 / metabolism
  • Phosphopyruvate Hydratase / genetics
  • Phosphopyruvate Hydratase / metabolism
  • Promoter Regions, Genetic
  • Proteoglycans / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Time Factors
  • Transduction, Genetic / methods
  • Transgenes / physiology*
  • Ubiquitins / genetics
  • Ubiquitins / metabolism
  • Viral Envelope Proteins / genetics
  • Viral Envelope Proteins / metabolism

Substances

  • Antigens
  • G protein, vesicular stomatitis virus
  • Glial Fibrillary Acidic Protein
  • Luminescent Proteins
  • Membrane Glycoproteins
  • Peptide Elongation Factor 1
  • Proteoglycans
  • Ubiquitins
  • Viral Envelope Proteins
  • chondroitin sulfate proteoglycan 4
  • Green Fluorescent Proteins
  • Phosphopyruvate Hydratase