Nerve growth factor receptor-mediated gene transfer.
One obstacle to effective gene therapies for neurological disorders lies in the cell-type diversity of the nervous system, making it difficult to direct gene delivery vectors to specific types of cells. To meet this challenge, we have developed a recombinant peptide-based gene delivery vector that targets nerve growth factor (NGF) receptors. The peptide comprises a cell-targeting domain derived from the NGF hairpin motif containing loops 1 and 2 linked to a DNA-binding domain composed of SPKR repeats. In PC12 cells, it activated the high-affinity NGF receptor, TrkA, and displayed NGF-like bioactivity by promoting neurite outgrowth and cell survival after serum deprivation. When combined with a low molecular weight of polyethylenimine (PEI), the peptide condensed plasmid DNA into nanoparticles that efficiently transferred exogenous genes into PC12 cells, enhancing reporter gene expression 5600-fold over peptide-free DNA/PEI complexes. Co-incubation with free NGF inhibited this effect. Furthermore, the peptide enhanced gene expression in NGF-receptor-rich rat primary cortex neurons but not glial cells. An in vivo experiment targeting TrkA-expressing dorsal root ganglia demonstrated that the peptide-containing complexes were 9- to 14-fold more efficient in transfection than controls. These properties make the chimeric peptide a promising gene delivery vector for targeting specific subtypes of neurons.
Ma
Nan
N
Institute of Bioengineering and Nanotechnology, Singapore.
Wu
Shan Shan
SS
Ma
Yue Xia
YX
Wang
Xu
X
Zeng
Jieming
J
Tong
Guping
G
Huang
Yan
Y
Wang
Shu
S
D020816:Amino Acid Motifs; D000818:Animals; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005727:Ganglia, Spinal / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D008297:Male; D020932:Nerve Growth Factor / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D009928:Organ Specificity; D016716:PC12 Cells; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D020917:Receptor, trkA / Q000378:metabolism; D017475:Receptors, Nerve Growth Factor / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D004268:DNA-Binding Proteins; D010446:Peptide Fragments; D017475:Receptors, Nerve Growth Factor; D011993:Recombinant Fusion Proteins; D020932:Nerve Growth Factor; D020917:Receptor, trkA