Poly(2-Propylacrylic Acid) Increases In Vitro Bioactivity of Chitosan/mRNA Nanoparticles

Etienne Jeandupeux; Mohamad-Gabriel Alameh; Majed Ghattas; Gregory De Crescenzo; Marc Lavertu

doi:10.1016/j.xphs.2021.06.003

Poly(2-Propylacrylic Acid) Increases In Vitro Bioactivity of Chitosan/mRNA Nanoparticles

J Pharm Sci. 2021 Oct;110(10):3439-3449. doi: 10.1016/j.xphs.2021.06.003. Epub 2021 Jun 4.

Authors

Etienne Jeandupeux¹, Mohamad-Gabriel Alameh², Majed Ghattas¹, Gregory De Crescenzo³, Marc Lavertu⁴

Affiliations

¹ Polytechnique Montreal, Institute of Biomedical Engineering, Montreal, QC, Canada.
² Perelman School of Medicine, University of Pennsylvania, Philadelphia, USA.
³ Polytechnique Montreal, Institute of Biomedical Engineering, Montreal, QC, Canada; Polytechnique Montreal, Department of Chemical Engineering, Montreal, QC, Canada.
⁴ Polytechnique Montreal, Institute of Biomedical Engineering, Montreal, QC, Canada; Polytechnique Montreal, Department of Chemical Engineering, Montreal, QC, Canada. Electronic address: marc.lavertu@polymtl.ca.

PMID: 34090900
DOI: 10.1016/j.xphs.2021.06.003

Abstract

Chitosan-based nanoparticles have been extensively studied for the delivery of nucleic acids. Previous results suggest that these nanoparticles have limited ability to escape the endosome, one of the main cellular barriers hindering nucleic acid delivery. Escape can be improved by the addition of endosomolytic agents during the formulation process or by developing delivery systems with intrinsic properties to disrupt endosomal membranes. In this study, Poly(2-Propylacrylic Acid) (PPAA), an anionic synthetic polymer with known membrane lytic activity was added to the binary chitosan/mRNA nanoparticles to improve bioactivity. The ionization behavior of PPAA was characterized to identify conditions in which PPAA is sufficiently charged to interact electrostatically with chitosan and thus form nanoparticles. The physicochemical characteristics (hydrodynamic diameter, polydispersity index, ζ-potential) and the in vitro transfection efficiency (bioactivity) of this new family of CS/mRNA/PPAA ternary nanoparticles were evaluated. The addition of PPAA to CS/mRNA nanoparticles was shown to be an efficient strategy to augment in vitro bioactivity. The optimal formulation reached an expression level ~86% of the commercial lipid control at pH 6.5 without any signs of metabolic toxicity. In this paper, we report the effect of salt and pH on the ionization behavior of PPAA and demonstrate 1) successful incorporation of PPAA into/onto nanoparticles, 2) improved bioactivity with PPAA, and 3) that the kosmotropic effects of trehalose play a minimal role in the apparent increase in bioactivity in presence of trehalose.

Keywords: Chitosan; Gene delivery; Poly (2-propylacrylic acid); Trehalose; mRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acrylic Resins
Chitosan*
Nanoparticles*
RNA, Messenger / genetics

Substances

Acrylic Resins
RNA, Messenger
poly(2-propylacrylic acid)
Chitosan