Chitosan-based gene delivery vectors targeted to the peripheral nervous system.
A non-toxic, targeted, simple and efficient system that can specifically transfect peripheral sensorial neurons can pave the way towards the development of new therapeutics for the treatment of peripheral neuropathies. In this study chitosan (CH), a biodegradable polymer, was used as the starting material in the design of a multicomponent vector targeted to the peripheral nervous system (PNS). Polycation-DNA complexes were optimized using imidazole- and thiol-grafted CH (CHimiSH), in order to increase transfection efficiency and allow the formation of ligand conjugated nanocomplexes, respectively. The 50 kDa non-toxic fragment from the tetanus toxin (HC), shown to interact specifically with peripheral neurons and undergo retrograde transport, was grafted to the binary complex via a bi-functional poly(ethylene glycol) (HC-PEG) reactive for the thiol moieties present in the complex surface. The targeting of the developed nanocomplexes was assessed by means of internalization and transfection studies in the ND7/23 (neuronal) vs. NIH 3T3 (fibroblast) cell lines. Targeted transfection was further confirmed in dorsal root ganglion dissociated primary cultures. A versatile, multi-component nanoparticle system that successfully targets and transfects neuronal cell lines, as well as dorsal root ganglia (DRG) primary neuron cultures was obtained for the 1.0 (w/w) HC-PEG/DNA formulation.
Oliveira
Hugo
H
INEB, Instituto de Engenharia Biomédica, Divisão de Biomateriais, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal.
Pires
Liliana R
LR
Fernandez
Ramon
R
Martins
M Cristina L
MC
Simões
Sérgio
S
Pêgo
Ana P
AP
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002412:Cations / Q000737:chemistry; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008422:Materials Testing; D051379:Mice; D008969:Molecular Sequence Data; D015394:Molecular Structure; D041681:NIH 3T3 Cells; D053758:Nanoparticles / Q000737:chemistry; D017933:Peripheral Nervous System / Q000502:physiology*; D011092:Polyethylene Glycols / Q000737:chemistry / Q000378:metabolism; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection / Q000379:methods
D002412:Cations; D011092:Polyethylene Glycols; D004247:DNA; D048271:Chitosan