Bioinspired Star-Shaped Poly(L-Lysine) Polypeptides.pdf (1.65 MB)
Bioinspired Star-Shaped Poly(L-Lysine) Polypeptides; 2 Efficient Polymeric Nanocarriers for the Delivery of DNA 3 to Mesenchymal Stem Cells
journal contribution
posted on 2020-03-24, 16:17 authored by David Walsh, Robert MurphyRobert Murphy, Angela Panarella, Rosanne Raftery, Brenton CavanaghBrenton Cavanagh, Jeremy C. Simpson, Fergal O'BrienFergal O'Brien, Andreas HeiseAndreas Heise, Sally-Ann CryanSally-Ann CryanThe field of tissue engineering is increasingly recognizing that gene
therapy can be employed for modulating in vivo cellular response thereby
guiding tissue regeneration. However, the field lacks a versatile and
biocompatible gene delivery platform capable of efficiently delivering
transgenes to mesenchymal stem cells (MSCs), a cell type often
refractory to transfection. Herein, we describe the extensive and
systematic exploration of three architectural variations of star-shaped
poly(l-lysine) polypeptide (star-PLL) with varying number and length of
poly(l-lysine) arms as potential nonviral gene delivery vectors for
MSCs. We demonstrate that star-PLL vectors are capable of
self-assembling with pDNA to form stable, cationic nanomedicines.
Utilizing high content screening, live cell imaging, and mechanistic
uptake studies we confirm the intracellular delivery of pDNA by
star-PLLs to MSCs is a rapid process, which likely proceeds via a
clathrin-independent mechanism. We identify a star-PLL composition with
64 poly(l-lysine) arms and five l-lysine subunits per arm as a
particularly efficient vector that is capable of delivering both
reporter genes and the therapeutic transgenes bone morphogenetic
protein-2 and vascular endothelial growth factor to MSCs. This
composition facilitated a 1000-fold increase in transgene expression in
MSCs compared to its linear analogue, linear poly(l-lysine).
Furthermore, it demonstrated comparable transgene expression to the
widely used vector polyethylenimine using a lower pDNA dose with
significantly less cytotoxicity. Overall, this study illustrates the
ability of the star-PLL vectors to facilitate efficient, nontoxic
nucleic acid delivery to MSCs thereby functioning as an innovative
nanomedicine platform for tissue engineering applications.