Ionizable lipid nanoparticles for in utero mRNA delivery

Abstract

Clinical advances enable the prenatal diagnosis of genetic diseases that are candidates for gene and enzyme therapies such as messenger RNA (mRNA)–mediated protein replacement. Prenatal mRNA therapies can treat disease before the onset of irreversible pathology with high therapeutic efficacy and safety due to the small fetal size, immature immune system, and abundance of progenitor cells. However, the development of nonviral platforms for prenatal delivery is nascent. We developed a library of ionizable lipid nanoparticles (LNPs) for in utero mRNA delivery to mouse fetuses. We screened LNPs for luciferase mRNA delivery and identified formulations that accumulate within fetal livers, lungs, and intestines with higher efficiency and safety compared to benchmark delivery systems, DLin-MC3-DMA and jetPEI. We demonstrate that LNPs can deliver mRNAs to induce hepatic production of therapeutic secreted proteins. These LNPs may provide a platform for in utero mRNA delivery for protein replacement and gene editing.

Lipid nanoparticles for mRNA delivery

“…In addition, lipid nanoparticle–mRNA formulations encoding immune stimulators can be directly delivered into tumour tissue by intratumoural injection 106 , 161 – 163 , to boost a local pro-inflammatory environment, which leads to immune cell activation and subsequent priming of systemic anticancer responses 106 , 161 – 163 . Finally, in utero administration of lipid nanoparticle–mRNA formulations can be applied to deliver mRNA to mouse fetuses 164 , achieving protein expression in fetal livers, lungs and intestines 164 .…”

Time evolution of PEG-shedding and serum protein coronation determines the cell uptake kinetics and delivery of lipid nanoparticle formulated mRNA

“…[5,[15][16][17][18] LNPs are typically comprised of an ionizable cationic lipid to encapsulate RNA and aid intracellular delivery with low toxicity, [19,20] combined with several "helper lipids", [21] such as phospholipids, a sterol, and a lipidated polyethylene glycol (PEG; PEG-ylated lipid). In addition to the introduction of new ionizable lipids, [17,[22][23][24][25][26][27] LNP efficacy can be significantly modulated by fine-tuning of LNP lipid compositions to induce changes of physicochemical parameters such as size and surface properties [28] or by chemical modification of individual helper components such as cholesterol. [29] Optimisation can result in both improved total cell uptake and, importantly, better endosomal escape.…”

Lipid in Chips: A Brief Review of Liposomes Formation by Microfluidics

“… 104 In fact, commercial devices based on micromixers such as NanoAssemblr™ platforms have been universally used in many studies to prepare liposome/LNPs for different applications, such as CRISPR-Cas9 genome editing for cancer therapy 105 and in utero mRNA delivery for monogenic fetal diseases. 106 Notably, COVID-19 vaccine nanoparticles manufactured by Pfizer are reported to be prepared by microfluidic mixers. 93 …”