title: Degradable self-assembling dendrons for gene delivery: experimental and theoretical insights into the barriers to cellular uptake.
abstract: This paper uses a combined experimental and theoretical approach to gain unique insight into gene delivery. We report the synthesis and investigation of a new family of second-generation dendrons with four triamine surface ligands capable of binding to DNA, degradable aliphatic-ester dendritic scaffolds, and hydrophobic units at their focal points. Dendron self-assembly significantly enhances DNA binding as monitored by a range of experimental methods and confirmed by multiscale modeling. Cellular uptake studies indicate that some of these dendrons are highly effective at transporting DNA into cells (ca. 10 times better than poly(ethyleneimine), PEI). However, levels of transgene expression are relatively low (ca. 10% of PEI). This indicates that these dendrons cannot navigate all of the intracellular barriers to gene delivery. The addition of chloroquine indicates that endosomal escape is not the limiting factor in this case, and it is shown, both experimentally and theoretically, that gene delivery can be correlated with the ability of the dendron assemblies to release DNA. Mass spectrometric assays demonstrate that the dendrons, as intended, do degrade under biologically relevant conditions over a period of hours. Multiscale modeling of degraded dendron structures suggests that complete dendron degradation would be required for DNA release. Importantly, in the presence of the lower pH associated with endosomes, or when bound to DNA, complete degradation of these dendrons becomes ineffective on the transfection time scale-we propose this explains the poor transfection performance of these dendrons. As such, this paper demonstrates that taking this kind of multidisciplinary approach can yield a fundamental insight into the way in which dendrons can navigate barriers to cellular uptake. Lessons learned from this work will inform future dendron design for enhanced gene delivery.
authors: Department of Chemistry, University of York, Heslington, York, YO10 5DD, UK.
authors: Anna
authors: A
authors: Barnard
authors: Paola
authors: P
authors: Posocco
authors: Sabrina
authors: S
authors: Pricl
authors: Marcelo
authors: M
authors: Calderon
authors: Rainer
authors: R
authors: Haag
authors: Mark E
authors: ME
authors: Hwang
authors: Victor W T
authors: VW
authors: Shum
authors: Daniel W
authors: DW
authors: Pack
authors: David K
authors: DK
authors: Smith
mesh_terms: D000873:Anthracenes; D004247:DNA / Q000378:metabolism; D018014:Gene Transfer Techniques; D013058:Mass Spectrometry; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D011094:Polyethyleneimine / Q000378:metabolism; D019076:Transgenes
chemical_list: D000873:Anthracenes; C501244:dendron; D011094:Polyethyleneimine; D004247:DNA