AAV-mediated targeting of gene expression to the peri-infarct region in rat cortical stroke model

J Neurosci Methods. 2014 Oct 30:236:107-13. doi: 10.1016/j.jneumeth.2014.08.014. Epub 2014 Aug 23.

Abstract

Background: For stroke patients the recovery of cognitive and behavioral functions is often incomplete. Functional recovery is thought to be mediated largely by connectivity rearrangements in the peri-infarct region. A method for manipulating gene expression in this region would be useful for identifying new recovery-enhancing treatments.

New method: We have characterized a way of targeting adeno-associated virus (AAV) vectors to the peri-infarct region of cortical ischemic lesion in rats 2days after middle cerebral artery occlusion (MCAo).

Results: We used magnetic resonance imaging (MRI) to show that the altered properties of post-ischemic brain tissue facilitate the spreading of intrastriatally injected nanoparticles toward the infarct. We show that subcortical injection of green fluorescent protein-encoding dsAAV7-GFP resulted in transduction of cells in and around the white matter tract underlying the lesion, and in the cortex proximal to the lesion. A similar result was achieved with dsAAV7 vector encoding the cerebral dopamine neurotrophic factor (CDNF), a protein with therapeutic potential.

Comparison with existing methods: Viral vector-mediated intracerebral gene delivery has been used before in rodent models of ischemic injury. However, the method of targeting gene expression to the peri-infarct region, after the initial phase of ischemic cell death, has not been described before.

Conclusions: We demonstrate a straightforward and robust way to target AAV vector-mediated over-expression of genes to the peri-infarct region in a rat stroke model. This method will be useful for studying the action of specific proteins in peri-infarct region during the recovery process.

Keywords: Adeno-associated virus; Animal models; Focal ischemia; Functional recovery; Gene transfer; Peri-infarct region.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / pathology
  • Brain / physiopathology*
  • Brain / virology
  • Brain Ischemia / pathology
  • Brain Ischemia / physiopathology
  • Brain Ischemia / therapy
  • Brain Ischemia / virology
  • Dependovirus / genetics*
  • Disease Models, Animal
  • Gene Expression*
  • Genetic Therapy / methods
  • Genetic Vectors*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Immunohistochemistry
  • Infarction, Middle Cerebral Artery
  • Magnetic Resonance Imaging
  • Male
  • Metal Nanoparticles
  • Nerve Growth Factors / genetics
  • Nerve Growth Factors / metabolism
  • Rats, Sprague-Dawley
  • Stroke / pathology
  • Stroke / physiopathology*
  • Stroke / therapy
  • Stroke / virology
  • Transduction, Genetic / methods

Substances

  • CDNF protein, rat
  • Nerve Growth Factors
  • Green Fluorescent Proteins