On the mechanism of tissue-selective gene delivery by lipid nanoparticles

The era of nucleic acid nanomedicine has arrived, as evidenced by Patisiran, a small interfering RNA (siRNA) encapsulated lipid nanoparticle (LNP), and mRNA-loaded LNPs used in COVID-19 vaccines. The diversity of nano-designs for delivering nucleic acid molecules tested in Phase II/III clinical trials reflects the potential of these technologies. These breakthroughs in non-viral gene delivery, including the use of LNPs, have attracted substantial interest worldwide for developing more effective drugs. A next step in this field is to target tissues other than the liver, which requires significant research efforts and material development. However, mechanistic studies in this area are lacking. This study compares two types of LNPs with different tissue-selectivity for delivering plasmid DNA (pDNA), one being liver-selective and the other spleen-selective, in an effort to understand the mechanisms responsible for differences in gene expression of delivered genes. We observed little difference in the biodistribution of these two LNPs despite the 100–1000-fold differences in gene expression. We then quantified the amount of delivered pDNA and mRNA expression in each tissue by quantitative real-time PCR (qPCR) to evaluate various intracellular processes, such as nuclear delivery, transcription and translation. The results showed a >100-fold difference in the translation step but there were little differences in amount of pDNA delivered to the nucleus or the amount of mRNA expression for the two LNP deliveries. Our findings suggest that endogenous factors affect gene expression efficiency not the extent of biodistribution. Biodistribution is necessary but not sufficient for achieving functional gene delivery, and we need to explore where the LNP transfection affects the biological process that influences protein expression. [Display omitted] •Two types of LNPs containing different helper lipids showed different tissue-selectivity for functional in vivo gene delivery.•The translational process appears to be a dominant step for causing the difference in gene expression of the delivered genes.•Transcriptional states were different between gene-expressing cells and low/non-expressing cells.•Endogenous factors might affect, not only the biodistribution, but also subsequent intracellular processes related to functional delivery.