abstract
The efficacy of brain therapeutics is largely hampered by the presence of the blood-brain barrier (BBB), mainly due to the failure of most (bio) pharmaceuticals to cross it. Accordingly, this study aims to develop nanocarriers for targeted delivery of recombinant precursor microRNA (pre-miR-29b), foreseeing a decrease in the expression of the BACE1 protein, with potential implications in Alzheimer's disease (AD) treatment. Stearic acid (SA) and lactoferrin (Lf) were successfully exploited as brain-targeting ligands to modify cationic polymers (chitosan (CS) or polyethyleneimine (PEI)), and its BBB penetration behavior was evaluated. The intracellular uptake of the dual-targeting drug delivery systems by neuronal cell models, as well as the gene silencing efficiency of recombinant pre-miR-29b, was analyzed in vitro. Labeled pre-miR-29b-CS/PEI-SA-Lf systems showed very strong fluorescence in the cytoplasm and nucleus of RBE4 cells, being verified the delivery of pre-miR-29b to neuronal cells after 1 h transfection. The experiment of transport across the BBB showed that CS-SA-Lf delivered 65% of recombinant pre-miR-29b in a period of 4 h, a significantly higher transport ratio than the 42% found for PEI-SA-Lf in the same time frame. Overall, a novel procedure for the dual targeting of DDS is disclosed, opening new perspectives in nanomedicines delivery, whereby a novel drug delivery system harvests the merits and properties of the different immobilized ligands.
keywords
TRANSFERRIN RECEPTOR; ALZHEIMERS-DISEASE; ENDOTHELIAL-CELLS; IN-VITRO; DRUG-DELIVERY; BARRIER; CHITOSAN; NANOPARTICLES; TRANSCYTOSIS; LIPOSOMES
subject category
Pharmacology & Pharmacy
authors
Pereira, P; Barreira, M; Cruz, C; Tomas, J; Luis, A; Pedro, AQ; Queiroz, JA; Sousa, F
our authors
Augusto Pedro
ResearcherProjects
Ionic liquid-based supports for pre-miRNAs purification targeting Alzheimer's disease (PUREmiRSILs)
Rede Nacional de Ressonância Magnética Nuclear (PTNMR)
acknowledgements
This work was partially supported by the Portuguese Foundation for Science and Technology (FCT), through the project Pest-OE/SAU/UI0709/2014, UIDB/50011/2020 and UIDP/50011/2020, UIDB/00285/2020, and by the project PTDC/BII-BBF/29496/2017 (PUREmiRSILs) funded by FEDER, through the COMPETE2020-Programa Operacional Competitividade e Internacionalizacao (POCI), and by national funds (OE), through FCT/MCTES. The authors also acknowledge the PPBI-Portuguese Platform of BioImaging through the project POCI-01-0145-FEDER-022122 and Rede Nacional de RMN (PTNMR), supported by FCT-MCTES (ROTEIRO/0031/2013-PINFRA/22161/2016) co-funded by FEDER through COMPETE 2020, POCI, and PORL, and FCT through PIDDAC.