Delineating effect of cationic head group and preparation method on transfection versus toxicity of lipid-based nanoparticles for gene delivery

Abstract Gene therapy using plasmid DNA (pDNA) is well-explored for variety of genetic diseases. However, its susceptibility to enzymatic degradation desires an optimal delivery system for efficient cellular uptake, transfection, and stability in vivo . Non-viral vectors like lipoplexes and LNPs have gained traction but there is no comparative evaluation of these lipid nanocarriers to deliver pDNA. Here, we demonstrated parallel comparison of both formulation components and technology for proficient pDNA delivery. Cationic and ionizable head groups were screened to find balance between acceptable transfection efficiency and systemic safety to deliver GFP-pDNA. We observed that lipoplexes formulated using SM-102 as biodegradable ionizable lipid exhibited high transfection efficiency given their high cellular uptake in A375V cells. Ionizable LNPs were fabricated via microfluidics and systematic comparison of lipid nanocarrier with GFP-pDNA complexed on interior versus exterior of nanoparticles was executed. We found LNPs to unveil high transfection efficiency and penetration within 3D spheroid model, while protecting pDNA under simulated physiological conditions. Our study lays a foundation to opt for the right complexing lipid and technology for development of lipid nanocarriers. Taken together, our research has opened the doors to designing “state-of-the-art” LNP based therapies by entrapping any functional plasmid gene that target life-threatening ailments.