Diblock Copolymer Targeted Lipid Nanoparticles: Next-Generation Nucleic Acid Delivery System Produced by Confined Impinging Jet Mixers

📖 Top 20% JournalOct 31, 2024ACS applied bio materials

Targeted Lipid Nanoparticles Made by Controlled Mixing for Improved Gene Delivery

AI simplified

mRNA Technology on OpenScience ↗PubMed ↗DOI ↗

Abstract

BCP-LNPs show up to a 40-fold increase in transfection efficiency in Hela cells compared to previous formulations.

The rapid desorption of lipid-PEG in the bloodstream limits the extrahepatic delivery of nucleic acids.
Biodegradable block copolymers (BCPs) may enhance the stability and targeting of lipid nanoparticles.
Optimal formulations for BCP-stabilized lipid nanoparticles (BCP-LNPs) were identified, achieving desirable properties and high encapsulation efficiency.
Certain BCP-LNP formulations demonstrated minimal cytotoxicity while significantly improving transfection rates.
BCP-LNPs successfully encapsulate and deliver both mRNA and pDNA, indicating a versatile cargo-agnostic nature.
Targeted BCP-LNPs conjugated with Cetuximab exhibit up to 2.4 times higher transfection in EGFR-overexpressing A549 cells compared to nontargeted LNPs.

AI simplified

Despite the recent advances and clinical demonstration of lipid nanoparticles (LNPs) for therapeutic and prophylactic applications, the extrahepatic delivery of nucleic acids remains a significant challenge in the field. This limitation arises from the rapid desorption of lipid-PEG in the bloodstream and clearance to the liver, which hinders extrahepatic delivery. In response, we explore the substitution of lipid-PEG with biodegradable block copolymers (BCPs), specifically poly(ε-caprolactone)--poly(ethylene glycol) (PCL--PEG). BCPs offer strong anchoring for large macromolecules, potentially enhancing cell-specific targeting. To develop and optimize BCP-stabilized LNPs (BCP-LNPs), we employed a Design of Experiment (DOE) approach. Through a systematic exploration, we identified optimal formulations for BCP-LNPs, achieving desirable physicochemical properties and encapsulation efficiency. Notably, BCP-LNPs exhibit surprising trends in transfection efficiency, with certain formulations showing up to a 40-fold increase in transfection in Hela cells, while maintaining minimal cytotoxicity. The lipid compositions that optimized PCL--PEG LNP transfection were different from the compositions that optimized PEG-lipid LNP transfection. Furthermore, our study confirms the versatility of BCP-LNPs in encapsulating and delivering both mRNA and pDNA, demonstrating their cargo-agnostic nature. Lastly, we showcased the targeted BCP-LNPs using a Cetuximab-conjugated formulation. These targeted LNPs show significant promise in delivering cargo specific to EGFR-overexpressing cells (A549 cells), with up to 2.4 times higher transfection compared to nontargeted LNPs. This finding underscores the potential of BCP-LNPs in targeted gene therapy, especially in challenging scenarios such as tumor targeting. Overall, our study establishes the viability of BCP-LNPs as a versatile, efficient, and targeted delivery platform for nucleic acids, opening avenues for advanced therapeutic applications. block b b

Full Text

Full text is available at the source.

View full text (PDF) ↗View full text ↗