Controlled proliferation of neural progenitor cells (NPCs) is a potential strategy for the treatment of neurodegenerative conditions such as Huntington's Disease and stroke. The subventricular zone (SVZ) of the adult mammalian brain harbors an NPC population that has been shown to migrate throughout the brain and mature into new neurons. It has been shown that stimulating the proliferation and differentiation of NPCs through the delivery of therapeutic genes encoding mitogenic factors such as fibroblast growth factor 2 (FGF2) can cause increased recruitment to damaged areas in the brain. The work described herein proposes to develop a multi-component nonviral gene delivery vehicle for FGF2 delivery to NPCs of the SVZ. The delivery vehicle will incorporate two bioactive peptides: (1) a targeting peptide to maximize uptake of particles and (2) a lytic peptide to mediate endosomal escape. In addition, vehicles will be modified with polyethylene glycol (PEG) for salt stability, which is required for vehicles administered in vivo. The proposed work is outlined in the aims below:
Aim 1 : Vehicles that incorporate both Tet1, HGP, and PEG will be synthesized. Vehicle formulations will be screened for desired physicochemicaj properties and optimized for transfection efficiency and binding in vitro. Materials will also be tested for toxicity using a cell viability assay.
Aim 2 : Optimal plasmid and polymer amount will be determined in vivo by intraventricular administration of vehicles. Formulations from Aim 1 will be used to deliver reporter gene constructs. Bulk expression will be quantified in brain lysate and distribution of expression will be determined by immunolabeling of brain slices.
Aim 3 : The optimal vehicle found in Aim 2 will be used to deliver FGF2. Quantification and localization of FGF2 expression will be done using ELISA and immunolabeling. FGF2-mediated proliferation and differentiation in brain slices will be identified by BrdU+/NeuN+ labeling and quantified by serology. This research investigates a potential strategy for the treatment of neurodegenerative diseases. It has relevance to public health since neurodegenerative diseases affects over 20 million individuals worldwide.
| Kwon, Ester J; Liong, Sylvie; Pun, Suzie H (2010) A truncated HGP peptide sequence that retains endosomolytic activity and improves gene delivery efficiencies. Mol Pharm 7:1260-5 |
| Kwon, Ester J; Lasiene, Jurate; Jacobson, Berit E et al. (2010) Targeted nonviral delivery vehicles to neural progenitor cells in the mouse subventricular zone. Biomaterials 31:2417-24 |
