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- W2896232032 abstract "Gene therapy has become a promising approach for neurodegenerative disease treatment, but there is still a lot to achieve in the development of an efficient gene carrier. In this study, we strategically designed a dual functionalized liposomes for efficient neuronal transfection by combining transferrin (Tf) receptor targeting, enhanced cell penetration utilizing penetratin (Pen) and gene protection against enzymatic degradation. Liposomal nanoparticles were prepared using thin lipid film hydration method while dual-functionalized liposomes were prepared by incorporating Tf-micelles into Pen-liposomes by post-insertion technique as previously reported. Biodistribution and biocompatibility were studied in C57BL/6J mice using DiR-labeled liposomes (excitation: 748 nm and emission: 782 nm) via tail vein injection at a dose of ∼15.2 μmoles phospholipids/kg body weight. After 24 h, ex vivo fluorescent images were taken by near infrared (NIR) imaging. In vivo gene expression was studied by single dose of liposomal formulations (∼15.2 μmoles phospholipids/kg body weight) loading pβ-gal (30 μg/mouse) or buffer containing pβgal or only buffer. Immunofluorescenece studies was conducted by injecting single dose of liposomal formulations (∼15.2 μmoles phospholipids/kg body weight) loading pGFP (30 μg/mouse). After 7 days, the brains were embedded in OCT compound and snap frozen in the liquid nitrogen. Tissue sections (30 μm thick) were cut using cryostat, fixed in acetone and methanol and incubated with primary antibody (anti-GFP antibody 1:100, Invitrogen) at 4°C overnight. Thereafter, sections were incubated with Alexa Fluor 488 secondary antibody diluted to 1:200 and observed by confocal microscope. A dual-functionalized liposome was developed to cross BBB and deliver gene to brain cells. The system could reach brain parenchyma and release pDNA in the cytoplasm of neurons, induce β-galactosidase production in vivo, thereby eliciting a better effect than the non-modified liposomes as well as naked pDNA. The study illustrated the superior ability of dual-functionalized liposomes to accumulate in the brain and transfect neurons. These results indicated that the liposomal formulation might be an efficient gene carrier and considered a promising approach for gene therapy of CNS diseases." @default.
- W2896232032 created "2018-10-26" @default.
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- W2896232032 date "2018-07-01" @default.
- W2896232032 modified "2023-10-16" @default.
- W2896232032 title "P4‐229: LIPOSOMAL NANOPARTICLES FOR GENE DELIVERY TO THE BRAIN" @default.
- W2896232032 doi "https://doi.org/10.1016/j.jalz.2018.07.050" @default.
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