Selective Targeting of Adenoviral Vectors to Neural Precursor Cells in the Hippocampus of Adult Mice: New Prospects for In Situ Gene Therapy

Abstract

The adult brain contains neural precursor cells (NPC) that are attracted to brain lesions, such as areas of neurodegeneration, ischemia, and cancer. This suggests that NPC engineered to promote lineage-specific differentiation or to express therapeutic genes might become a valuable tool for restorative cell therapy and for targeting therapeutic genes to diseased brain regions. Here we report the identification of NPC-specific ligands from phage display peptide libraries and show their potential to selectively direct adenovirusmediated gene transfer to NPC in adult mice. Identified peptides mediated specific virus binding and internalization to cultured neurospheres. Importantly, peptide-mediated adenoviral vector infection was restricted to precursor cells in the hippocampal dentate gyrus of pNestingreen fluorescent protein transgenic or C57BL/6 mice. Our approach represents a novel method for specific manipulation of NPC in the adult brain and may have major implications for the use of precursor cells as therapeutic delivery vehicles in the central nervous system.

Novel subventricular zone early progenitor cell-specific adenovirus for in vivo therapy of central nervous system disorders reinforces brain stem cell heterogeneity

“…Primary neural stem and progenitor cells (NSPCs) were isolated from the lateral wall of lateral ventricles (SVZ) and hippocampus (HC) of 4-week-old C57/B6 mice (Charles River Lab., USA), expanded, and differentiated under removal of growth factors as previously described (Schmidt et al , 2007. Mice were anaesthetized using ketamine (75 mg/kg) and rompun (5.8 mg/kg) and killed by decapitation.…”

“…Thus, the most important challenge is to develop Ad-derived gene delivery systems with high efficiency through target cell selectivity, in particular under in situ conditions (Reetz et al 2014). Our recent endeavors in the generation of Ad vectors that specifically target stem/progenitor cells in the body have focused on linking selective peptides to viral particles that direct the virus to specific or yet unknown receptors on the surface of stem cells (Hildebrandt et al 2010;Schmidt et al 2007). Using an Ad-PEGylation approach, we demonstrated that a short artificial peptide selected by phage display, which specifically binds neural precursor cells derived from the hippocampal area of adult mice or rats, was highly effective in targeting NSPCs after virus administration into the brain (Hildebrandt et al 2010;Schmidt et al 2007).…”

“…Our recent endeavors in the generation of Ad vectors that specifically target stem/progenitor cells in the body have focused on linking selective peptides to viral particles that direct the virus to specific or yet unknown receptors on the surface of stem cells (Hildebrandt et al 2010;Schmidt et al 2007). Using an Ad-PEGylation approach, we demonstrated that a short artificial peptide selected by phage display, which specifically binds neural precursor cells derived from the hippocampal area of adult mice or rats, was highly effective in targeting NSPCs after virus administration into the brain (Hildebrandt et al 2010;Schmidt et al 2007). These studies also revealed that longterm transgene expression was restricted to precursor cells located in dentate gyrus, while not detected in other injected brain regions such as the SVZ and the ependyma of the lateral ventricle.…”

Nanomedicine Approaches to Modulate Neural Stem Cells in Brain Repair

“…Additionally, nanomaterials can incorporate peptide ligands to specifically target endogenous NSCs. Some ligands have been identified by phage display peptide libraries [5]. Importantly, functionalized nanostructured hydrogels were shown to successfully induce NSC differentiation while supporting tissue regeneration [6].…”

Gene Delivery Systems: Tailoring Vectors to Reach Specific Tissues

“…A similar approach involves the pre-selection of peptides by using a phage display peptide library, from which peptides with high affinity for a certain cell type are selected and then inserted into the AdV vector capsid. This has allowed the targeting of AdV vectors to epithelium cells (Nicklin et al, 2001b), tumor cells (Rittner et al, 2007), and neurons (Schmidt et al, 2007). Most efforts to modify the AdV vector capsid have been on vector targeting to tumor cells (Bachtarzi et al, 2008).…”