# Aminoglycoside Drugs as Adjuvants to Enhance siRNA/mRNA Delivery by Lipid Nanoparticles

## TL;DR

This study shows that aminoglycoside drugs, like kanamycin, can help lipid nanoparticles deliver siRNA and mRNA more effectively by improving lysosomal escape.

## Contribution

The study introduces aminoglycosides as novel adjuvants to enhance nucleic acid delivery by lipid nanoparticles.

## Key findings

- Aminoglycosides significantly improved LNP/siRNA delivery efficiency, reducing luciferase expression to 22.15%.
- Kanamycin sulfate destabilized lysosomal membranes and reduced siRNA colocalization with lysosomes.
- Kanamycin enhanced silencing of EGFR mRNA and boosted spike protein expression from LNPs/mRNA.

## Abstract

Nucleic acid drugs (such as siRNA and mRNA) hold broad
prospects in disease treatment, but their clinical application is
limited by low delivery efficiency, particularly due to insufficient
lysosomal escape. This study aims to explore aminoglycoside small-molecule
drugs as adjuvants to enhance the lysosomal escape efficiency of lipid
nanoparticles (LNPs) for nucleic acid drug delivery, thereby improving
therapeutic effects and providing new strategies for gene therapy.
In this study, five FDA-approved aminoglycoside small molecules were
selected as adjuvants to enhance the lysosomal escape of LNPs/siRNA
and improve the siRNA delivery effect. In vitro results
confirmed that aminoglycoside drugs significantly enhanced the delivery
efficiency of LNPs/siRNA, reducing luciferase expression to as low
as 22.15%. Mechanistic studies revealed that kanamycin sulfate markedly
reduced siRNA colocalization with lysosomes and destabilized lysosomal
membranes under acidic conditions (pH 6.2), increasing hemolytic effects
by 3.6-fold. Additionally, kanamycin sulfate significantly improved
the silencing efficacy of LNPs/siEGFR on EGFR mRNA and protein expression
and boosted LNPs/mSARS-CoV-2-mediated spike protein expression. Therefore,
aminoglycoside small molecules (particularly kanamycin) can effectively
enhance nucleic acid delivery efficiency by disrupting endosomal membrane
structures, providing a novel adjuvant strategy for the clinical application
of siRNA and mRNA drugs.