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Download fileDelivery of Splice Switching Oligonucleotides by Amphiphilic Chitosan-Based Nanoparticles
journal contribution
posted on 2016-02-01, 00:00 authored by Pedro M. D. Moreno, Joyce C. Santos, Carla
P. Gomes, Aida Varela-Moreira, Artur Costa, Victoria Leiro, Herman Mansur, Ana P. PêgoSplice switching
oligonucleotides (SSOs) are a class of single-stranded
antisense oligonucleotides (ssONs) being used as gene therapeutics
and demonstrating great therapeutic potential. The availability of
biodegradable and biocompatible delivery vectors that could improve
delivery efficiencies, reduce dosage, and, in parallel, reduce toxicity
concerns could be advantageous for clinical translation. In this work
we explored the use of quaternized amphiphilic chitosan-based vectors
in nanocomplex formation and delivery of splice switching oligonucleotides
(SSO) into cells, while providing insights regarding cellular uptake
of such complexes. Results show that the chitosan amphiphilic character
is important when dealing with SSOs, greatly improving colloidal stability
under serum conditions, as analyzed by dynamic light scattering, and
enhancing cellular association. Nanocomplexes were found to follow
an endolysosomal route with a long lysosome residence time. Conjugation
of a hydrophobic moiety, stearic acid, to quaternized chitosan was
a necessary condition to achieve transfection, as an unmodified quaternary
chitosan was completely ineffective. We thus demonstrate that amphiphilic
quaternized chitosan is a biomaterial that holds promise and warrants
further development as a platform for SSO delivery strategies.
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Keywords
Splice Switching Oligonucleotidesamphiphilic quaternized chitosandelivery efficiencieslysosome residence timeserum conditionsgene therapeuticsquaternized chitosanstearic acidbiocompatible delivery vectorsquaternary chitosanendolysosomal routeSSO delivery strategieschitosan amphiphilic characteroligonucleotidenanocomplex formationtoxicity concernsResults show