Formulation of Lipid-Free Polymeric Mesoscale Nanoparticles Encapsulating mRNA

Abstract

Introduction Nanoparticle-mediated gene therapy has found substantial clinical impact, primarily focused on lipid-based nanoparticles. In comparison with lipid nanoparticles, polymeric particles may have certain advantages such as increased biocompatibility and controlled release. Our prior studies have found that polymeric mesoscale nanoparticles exhibited specific targeting to the renal proximal tubules. Thus, in this study, we sought to identify formulation parameters that allow for development of polymeric mesoscale nanoparticles encapsulating functional mRNA for delivery into tubular epithelial cells. Methods We evaluated particle uptake in vitro prior to exploring formulation parameters related to introduction of a primary mixture of polymer in acetonitrile and hydrophilic mRNA in water. Finally, we evaluated their functionality in a renal tubular epithelial cell line. Results We found that MNPs are endocytosed within 15 min and that the mesoscale nanoparticle formulation procedure was generally robust to introduction of a primary mixture and encapsulation of mRNA. These particles exhibited substantial uptake in renal cells in vitro and rapid (< 1 h) expression of a model mCherry fluorescent protein. Conclusion We anticipate these findings having potential in the delivery of specific gene therapies for renal disorders and cancer.

Targeting the Kidneys at the Nanoscale: Nanotechnology in Nephrology

“… 111 In addition, loading of mRNA into MNPs and expression of a reporter protein (mCherry) in renal tubular cells in vitro has been reported. 113 The ability to load and deliver functional nucleic acids to the kidneys, and generally anywhere beyond the liver, is an extremely important and exciting development in gene delivery science. The plug-and-play nature of siRNA and mRNA, coupled with recent developments in those fields, potentiates many therapeutic applications across a broad range of kidney diseases.…”

The clinical progress and challenges of mRNA vaccines

“…For polymers, it is easy to functionalize with various units, such as ligands (e.g., targeting peptide, antibody) for targeting, stimuli‐responsive materials that respond to an enzyme, ROS, and pH for controlled release of mRNA cargoes (Skelton et al, 2022; Ulkoski et al, 2021), which contribute to high translation of mRNA. Additionally, polymeric nanoparticles are usually composed of multifunctional polymers and will ensure stability during storage and shipping under freezing.…”