Efficient Nuclear Delivery and Nuclear Body Localization of Antisense Oligo-Nucleotides using Degradable Polymersomes

Abstract

Delivery of antisense oligonucleotides, AON, presents many of the same challenges as delivery of any nucleic acid: charge, stability, cell uptake, endolysosomal escape, and entry into the nucleus. Here we demonstrate efficient delivery of AON after loading into biodegradable polymer vesicles or 'polymersomes'. We focus on AON delivery to muscle cells in vitro and in vivo because of the emergence of AON in therapeutic strategies directed at muscular dystrophies. To first clarify uptake kinetics without the complications of typical multilayered myotube cultures, we use micro-patterned C2C12 cells and show efficient uptake of AONpolymersomes. The biodegradable polymersomes break down and foster AON escape with the binding of fluorescent-AON into the nuclear bodies. Intramuscular injections of the polymersome-AON into the hind limbs of mdx-dystrophic mice show more efficient nuclear uptake than AON alone and also lead to dystrophin expression in the mdx mice. In sum, these neutral, degradable carriers of AON show promise in vivo. This material is posted here with permission of the IEEE. Such permission of the IEEE does not in any way imply IEEE endorsement of any of the University of Pennsylvania's products or services. Internal or personal use of this material is permitted. However, permission to reprint/republish this material for advertising or promotional purposes or for creating new collective works for resale or redistribution must be obtained from the IEEE by writing to pubs-permissions@ieee.org. By choosing to view this document, you agree to all provisions of the copyright laws protecting it.

Polymersome‐mediated intracellular delivery of antibiotics to treat Porphyromonas gingivalis ‐infected oral epithelial cells

“…Being fully synthetic, the physiological properties of polymersomes can be easily manipulated to incorporate desirable characteristics that can have a dramatic effect on drug delivery (25, 45, 51). Polymersome‐delivered therapy has already been successfully tested in vivo (30–32); hence, it is perfectly feasible that in vitro studies such as that described herein can be replicated in vivo and rapidly translated into the clinic.…”

“…To date, polymersomes have been used to administer anticancer therapy in vitro (paclitaxel and doxorubicin) and to shrink tumors in vivo (29, 30). They have also been used for the intracellular delivery of siRNA and antisense oligonucleotides (31, 32) or functional antibodies (28) and as transfection tools to deliver plasmid DNA into cells (33, 34). In the current study we show, for the first time, that polmersomes can deliver antibiotics to oral keratinocytes and that, once delivered, these can kill intracellular‐dwelling bacteria.…”

Imaging and drug delivery using theranostic nanoparticles

“…A study demonstrated the specificity of an actively targeted polymeric vesicle via encapsulation and targeted release of a hydrophobic dye [77]. In addition, gene delivery has been demonstrated in vitro and in vivo , while simultaneously allowing molecular imaging of the polymersome [74,78,79]. Gd-DTPA and iron-oxide loaded PLGA nanoparticles have been used with US and MRI [80,81].…”