Targeted gene delivery into peripheral sensorial neurons mediated by self-assembled vectors composed of poly(ethylene imine) and tetanus toxin fragment c.
A simple, safe and efficient system that can specifically transfect peripheral sensorial neurons can bring new answers to address peripheral neuropathies. A multi-component non-viral gene delivery vector targeted to peripheral nervous system cells was developed, using poly(ethylene imine) (PEI) as starting material. A binary DNA/polymer complex based on thiolated PEI (PEISH) was optimized, considering complex size and zeta potential and the ability to transfect a sensorial neuron cell line (ND7/23). The 50 kDa non-toxic fragment from tetanus toxin (HC), which has been previously shown to interact specifically with peripheral neurons and to undergo retrograde transport, was grafted to the complex core via a bifunctional PEG (HC-PEG) reactive for the thiol moieties present in the complex surface. Several formulations of HC-PEG ternary complexes were tested for targeting, by assessing the extent of cellular internalization and levels of transfection, in both the ND7/23 and NIH 3 T3 (fibroblast) cell lines. Targeted gene transfer to the neuronal cell line was observed for the complex formulations containing 5 and 7.5 microg of HC-PEG. Finally, our results demonstrate that the developed ternary vectors are able to transfect primary cultures of dorsal root ganglion dissociated neurons in a targeted manner and elicit the expression of a relevant neurotrophic factor.
Oliveira
Hugo
H
INEB-Instituto de Engenharia Biomédica, Divisão de Biomateriais, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal.
Fernandez
Ramon
R
Pires
Liliana R
LR
Martins
M Cristina L
MC
Simões
Sérgio
S
Barbosa
Mário A
MA
Pêgo
Ana P
AP
D000818:Animals; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D004247:DNA / Q000008:administration & dosage*; D005260:Female; D007097:Imines / Q000737:chemistry*; D051379:Mice; D041681:NIH 3T3 Cells; D009474:Neurons / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry*; D017933:Peripheral Nervous System / Q000166:cytology*; D010957:Plasmids / Q000008:administration & dosage; D011095:Polyethylenes / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D013744:Tetanus Toxin / Q000737:chemistry*; D014162:Transfection
D007097:Imines; D010446:Peptide Fragments; D011095:Polyethylenes; D013744:Tetanus Toxin; C505405:poly(ethylene imine); C040635:tetanus toxin fragment C; D004247:DNA