Study the lipidoid nanoparticle mediated genome editing protein delivery using 3D intestinal tissue model

Area of Science:

• Biotechnology
• Nanomedicine
• Drug Delivery

Background:

• Lipid nanoparticles (LNPs) show potential for oral drug delivery.
• Maintaining cargo-carrier complex integrity after intestinal epithelial crossing is crucial for intracellular delivery of bioactive cargos like gene-editing proteins.
• Limited research exists on the integrity and intracellular delivery capabilities of LNP-protein nanocomplexes post-intestinal barrier transit.

Purpose of the Study:

• To evaluate the integrity and intracellular delivery efficacy of lipid nanoparticle (LNP)-protein nanocomplexes after crossing the intestinal epithelial barrier.
• To assess the utility of a 3D tissue-engineered intestine model for identifying effective oral drug delivery systems.

Main Methods:

• Utilized a traditional 2D transwell system and a 3D tissue-engineered intestine model.
• Encapsulated GFP-Cre recombinase (cargo) within EC16-63 LNPs (carrier).
• Assessed LNP-protein nanocomplex penetration of intestinal epithelial layers and subsequent intracellular delivery.

Main Results:

• EC16-63 LNPs efficiently encapsulated GFP-Cre recombinase.
• Nanocomplexes temporarily disrupted tight junctions to penetrate intestinal monolayers in both 2D and 3D models.
• Delivered cargo into underlying cells, inducing gene recombination post-transport across the intestinal epithelia.

Conclusions:

• In vitro 3D intestinal tissue models are valuable for screening effective lipid nanoparticles for oral delivery.
• EC16-63 LNP-protein nanocomplexes demonstrate successful cargo delivery across the intestinal barrier for intracellular gene editing applications.