Lipid nanoparticle topology regulates endosomal escape and delivery of RNA to the cytoplasm

Abstract

RNA therapeutics have the potential to resolve a myriad of diseases caused by gene deficiency. Lipid nanoparticles (LNPs) are one of the most successful RNA delivery systems. However, expanding their application hinges on the discovery of next generation LNPs with high potency, cyto-specific targeting, and low side effects. Overcoming the difficulty of releasing cargo from endocytosed LNPs remains a significant hurdle. The endosomal escape of viral and non-viral nanoparticles relies on the topological transformation of membrane fusion pore formation followed by RNA translocation into the cytosol. In this study we show that LNP-RNA nanostructure modulates the energetic cost of LNP fusion with a target membrane. The inclusion of a new class of structurally-active lipids leads to superior LNP endosomal fusion, fast evasion of endosomal entrapment, and efficacious RNA delivery. Specifically, bicontinuous cubic RNA-LNPs, cuboplexes, have significantly higher endosomal escape rates and deliver more RNA compared to regular lamellar LNPs.

Loading of extracellular vesicles with nucleic acids via hybridization with sponge-like lipid nanoparticles

“…Recent structural investigations on LNPs have shown that the structural organization of the lipid matrix can be modulated by the process parameters (e.g., buffer pH and automated vs. microfluidic preparation protocols) and the lipid composition (66)(67)(68). It was shown that LNPs with non-lamellar LLC domains (i.e., HII phases or the bicontinuous cubic phases (QII)) display improved transfections in vitro and in vivo (69,70). This was partially attributed to their enhanced fusogenic properties leading to increased endosomal escape and cytosolic NA delivery.…”

“…It was shown that LNPs with non-lamellar LLC domains (i.e. , H II phases or the bicontinuous cubic phases (Q II )) display improved transfections in vitro and in vivo ( 69, 70 ). This was partially attributed to their enhanced fusogenic properties leading to increased endosomal escape and cytosolic NA delivery.…”

Expanding CAR-T cell immunotherapy horizons through microfluidics

“…145 In addition, under acidic conditions within endosomes, ionizable LNPs acquire a positive charge, enabling efficient endosomal escape by destabilizing the endosomal membrane. 146 2.3.2.3.1. Microfluidic approach for LNP formation.…”

Advances in non-viral mRNA delivery to the spleen

“…79,84 This H II phase transition has been hypothesized to aid in the fusion of the lipid nanoparticle membrane to the endosomal membrane, thus facilitating endosomal escape. 85,86 Cholesterol is an additional component of lipid nanoparticles which has been…”