Gene delivery to differentiated neurotypic cells with RGD and HIV Tat peptide functionalized polymeric nanoparticles

Abstract

A number of neurodegenerative disorders may potentially be treated by the delivery of therapeutic genes to neurons. Nonviral gene delivery systems, however, typically provide low transfection efficiency in post-mitotic differentiated neurons. To uncover mechanistic reasons for this observation, we compared gene transfer to undifferentiated and differentiated SH-SY5Y cells using polyethylenimine (PEI)/DNA nanocomplexes. Differentiated cells exhibited substantially lower uptake of gene vectors. To overcome this bottleneck, RGD or HIV-1 Tat peptides were attached to PEI/DNA nanocomplexes via poly(ethylene glycol) (PEG) spacer molecules. Both RGD and Tat improved the cellular uptake of gene vectors and enhanced gene transfection efficiency of primary neurons up to 14-fold. RGD functionalization resulted in a statistically significant increase in vector escape from endosomes, suggesting it may improve gene delivery by more than one mechanism.

Rapid transport of large polymeric nanoparticles in fresh undiluted human mucus

“…The mechanism of particle transport over short and long time scales was classified based on the concept of relative change (RC) of D eff , as discussed (60,64). In brief, RC values of particles at short and long time scales were calculated by dividing the D eff of a particle at a probed time scale by the D eff at an earlier reference time scale.…”

Dual Responsive, Stabilized Nanoparticles for Efficient In Vivo Plasmid Delivery

“…[33] A main advantage of bPEI is its buffering capacity, which facilitates its endosomal escape. [34] To assess the proton buffering capacity (BC, μmol H + / mg polymer required to decrease the pH of a 0.2 mg/ml polymer solution from 7.4 to 5.0) of the various PCL-based copolymers, acid-base titration was performed ( Figure S3).…”

Brain Drug Delivery Systems for Neurodegenerative Disorders

“…However, a concern for gene therapy is the vector safety, as there is a risk of excessive immune response as well as insertional mutagenesis. Thus, the development of non-viral vectors is attractive due to the potential for improved safety, reduced immunogenicity, ease of manufacturing and scale-up, and the ability to accommodate larger DNA plasmids compared to viral vectors [55].…”

“…Within this line, researchers are assessing the feasibility of using nanotechnology to condense DNA plasmids into nanoparticles and deliver them to the brain as a means to halt or prevent the neurodegenerative process [55]. The neuroprotective effect of Lactoferrin (Lf)-modified nanoparticles encapsulating GDNF gene via a regimen of multiple dosing intravenous administration was examined in 2 different rat PD models.…”