Naturally-occurring cholesterol analogues in lipid nanoparticles induce polymorphic shape and enhance intracellular delivery of mRNA

Endosomal sequestration of lipid-based nanoparticles (LNPs) remains a formidable barrier to delivery. Herein, structure-activity analysis of cholesterol analogues reveals that incorporation of C-24 alkyl phytosterols into LNPs (eLNPs) enhances gene transfection and the length of alkyl tail, flexibility of sterol ring and polarity due to -OH group is required to maintain high transfection. Cryo-TEM displays a polyhedral shape for eLNPs compared to spherical LNPs, while x-ray scattering shows little disparity in internal structure. eLNPs exhibit higher cellular uptake and retention, potentially leading to a steady release from the endosomes over time. 3D single-particle tracking shows enhanced intracellular…

• CF
  Cystic Fibrosis Foundation

  Award: SAHAY18G0
• OH
  Oregon Health and Science University

  Award: P30CA069533
• M
  Moderna
• KC
  Knight Cancer Institute, Oregon Health and Science University

  Award: P30CA069533
• NH
  National Heart, Lung, and Blood Institute

  Award: 1R01HL146736-01
• NC
  National Cancer Institute

  Award: P30CA069533
• NI
  National Institute of Biomedical Imaging and Bioengineering

  Award: 1R15EB021581