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Poly(2-oxazoline)-Based Polyplexes as a PEG-Free Plasmid DNA Delivery Platform.

Dina N Yamaleyeva1,2, Naoki Makita2,3, Duhyeong Hwang2,4

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Macromolecular Bioscience
|July 19, 2023
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Summary

This study developed a polyethylene glycol (PEG)-free gene delivery platform using cationic poly(2-oxazoline) (POx) copolymers for immune cells. A specific DET- and pEtOx-based diblock copolymer showed superior transfection efficiency in macrophages, outperforming targeted versions and commercial reagents.

Keywords:
macrophagesmannose targetingpoly(2-oxazolines)transfection

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Area of Science:

  • Polymer chemistry and biomaterials science
  • Gene therapy and drug delivery
  • Cell biology and immunology

Background:

  • Polyethylene glycol (PEG) is widely used in gene delivery but can elicit immune responses.
  • Developing PEG-free gene delivery systems is crucial for improving safety and efficacy.
  • Cationic poly(2-oxazoline) (POx) copolymers offer a versatile platform for gene delivery applications.

Purpose of the Study:

  • To expand the utility of cationic POx copolymers as a PEG-free gene delivery platform for immune cells.
  • To synthesize and characterize novel POx-based block copolymers with varying hydrophilic, cationic, and hydrophobic blocks.
  • To evaluate the transfection efficiency of these copolymers in monocytes and macrophages, including targeted and non-targeted formulations.

Main Methods:

  • Synthesis of POx block copolymers with pMeOx or pEtOx hydrophilic blocks and modified MestOx cationic blocks (using DET or TREN).
  • Incorporation of a hydrophobic iPrOx block in triblock copolymers.
  • Functionalization with mannose targeting ligand via click chemistry.
  • Preparation of polyplexes and evaluation of transfection efficiency in macrophages and monocytes.
  • Assessment of cell internalization and comparison with a commercial transfection reagent (GeneJuice).

Main Results:

  • DET-containing copolymers showed significantly higher macrophage transfection than TREN-based ones.
  • Nontargeted pEtOx-based diblock copolymers were more effective than pMeOx-based counterparts.
  • Triblock copolymers with hydrophobic iPrOx blocks performed less effectively than diblock copolymers.
  • Mannose ligand attachment unexpectedly inhibited transfection and reduced cell internalization.
  • The optimized PEG-free, nontargeted DET- and pEtOx-based diblock copolymer achieved transfection levels comparable to GeneJuice.

Conclusions:

  • Cationic POx copolymers represent a promising PEG-free platform for gene delivery to immune cells.
  • The choice of hydrophilic block (pEtOx > pMeOx) and cationic side chain (DET > TREN) significantly impacts transfection efficiency.
  • Hydrophobic blocks can negatively affect performance, suggesting diblock structures are preferable.
  • Surface functionalization with mannose for targeting can be detrimental to transfection in this system.
  • A specific PEG-free, nontargeted POx diblock copolymer demonstrates high potential for macrophage gene delivery.