Synergistic lipid compositions for albumin receptor mediated delivery of mRNA to the liver

Abstract

Lipid-like nanoparticles (LNPs) have potential as non-viral delivery systems for mRNA therapies. However, repeated administrations of LNPs may lead to accumulation of delivery materials and associated toxicity. To address this challenge, we have developed biodegradable lipids which improve LNPs clearance and reduce toxicity. We modify the backbone structure of Dlin-MC3-DMA by introducing alkyne and ester groups into the lipid tails. We evaluate the performance of these lipids when co-formulated with other amine containing lipid-like materials. We demonstrate that these formulations synergistically facilitate robust mRNA delivery with improved tolerability after single and repeated administrations. We further identify albumin-associated macropinocytosis and endocytosis as an ApoEindependent LNP cellular uptake pathway in the liver. Separately, the inclusion of alkyne lipids significantly increases membrane fusion to enhance mRNA release, leading to synergistic improvement of mRNA delivery. We believe that the rational design of LNPs with multiple amine-lipids increases the material space for mRNA delivery.

Nanomaterial Delivery Systems for mRNA Vaccines

“…One of the challenges in this area is the lack of a reliable standardized endosomal release method that can be implemented broadly. Many methods have been developed, but are usually specific to only one lab group [ 42 , 101 , 145 , 146 , 147 , 148 , 149 ]. mRNA was also recently shown to undergo exocytosis in an amount that is similar to the amount released into the cytosol [ 150 ].…”

Lipid nanoparticle technology for therapeutic gene regulation in the liver

“…Efficient targeting and gene regulation in hepatocytes stems from the presence of ApoE in the corona of LNPs and enabled the success of Onpattro® [33,76]. A recent publication suggested that the ionizable lipid composition plays a major role in the corona formed [165]. How the biomolecular corona can be leveraged to optimize targeting of different cell types within the liver microenvironment needs to be investigated.…”

Advances in Lipid Nanoparticles for mRNA-Based Cancer Immunotherapy

“…Similarly, a biodegradable ionizable lipid obtained by modifying the hydrophobic tail of MC3 lipid through the introduction of ester and alkyne groups, in addition to showing an improved tolerability, displayed an enhanced transfection efficiency. In particular, Miao et al observed that the incorporation of an unsaturated alkyne group, rather than double bounds, in the non-polar tail of the ionizable lipid can improve the fusogenicity with the endosomal membrane, and consequently facilitate endosomal escape and mRNA release into the cytosol (Miao et al, 2020). Additionally, its co-formulation with cKK-E12 pH-responsive cationic lipid synergistically boosted mRNA delivery into hepatocytes, offering early evidence that novel and more efficient delivery systems could be potentially obtained from the co-formulation of distinct ionizable lipids.…”

“…Additionally, its co-formulation with cKK-E12 pH-responsive cationic lipid synergistically boosted mRNA delivery into hepatocytes, offering early evidence that novel and more efficient delivery systems could be potentially obtained from the co-formulation of distinct ionizable lipids. cKK-E12-based lipids have been shown to enhance the serum stability and protein binding of the particles (Miao et al, 2020).…”

“…Different types of nanoparticles have been proposed for mRNA delivery, but lipid-based nanoparticles have shown the most promising results and currently represent the gold standard for precise in vivo delivery of mRNA into immune cells (Kranz et al, 2016;Oberli et al, 2017;Persano et al, 2017;Miao et al, 2020). A proof-of-concept study using this approach was reported by Kranz et al in 2016, demonstrating that DCs can be passively targeted in vivo using intravenously injected mRNA-lipoplexes based on DOTMA/DOPE or DOTAP/DOPE formulations by optimizing the mRNA/cationic lipid ratio, obtaining nanoparticles with a negative net charge (Kranz et al, 2016).…”