Search

Lipid nanoparticles (LNPs) containing ionizable aminolipids are among the leading platforms for the successful delivery of nucleic-acid-based therapeutics, including messenger RNA (mRNA). The two recently FDA-approved COVID-19 vaccines developed by Moderna and Pfizer/BioNTech belong to this category. Ionizable aminolipids, cholesterol, and DSPC lipids are among the key components of such formulations, crucially modulating physicochemical properties of these formulations and, consequently, the potency of these therapeutics. Despite the importance of these components, the distribution of these molecules in LNPs containing mRNA is not clear. In this study, we used all-atom molecular dynamics (MD) simulations to investigate the distribution and effects of the Lipid-5 (apparent pKaof the lipid nanoparticle = 6.56), a rationally designed and previously reported ionizable aminolipid by Moderna, on lipid bilayers [Mol. Ther.2018, 26, 1509–1519]. The simulations were conducted with half of the aminolipids charged and half neutral approximately to the expected ionization in the microenvironment of the LNP surface. In all five simulated systems in this work, the cholesterol content was kept constant, whereas the DSPC and Lipid-5 concentrations were changed systematically. We found that at higher concentrations of the ionizable aminolipids, the neutral aminolipids form a disordered aggregate in the membrane interior that preferentially includes cholesterol. The rules underlying the lipid redistribution could be used to rationally choose lipids to optimize the LNP function.

Fengycins are a class of antifungal lipopeptides synthesized by the bacteria Bacillus subtilis, commercially available as the primary component of the agricultural fungicide Serenade. They are toxic to fungi but far less to mammalian cells. One key difference between mammalian and fungal cell membranes is the presence of cholesterol only in the former; recent experimental work showed that the presence of cholesterol reduces fengycin-induced membrane leakage (1). Since our previous all-atom and coarse-grained simulations suggested that aggregation of membrane-bound fengycin is central to its ability to disrupt membranes, we hypothesized that cholesterol might reduce fengycin aggregation (2,3). Here, we test this hypothesis using coarse-grained molecular dynamics simulations, with sampling enhanced via the weighted ensemble method. The results indicate that cholesterol subtly alters the size distribution for fengycin aggregates, limits the lateral range of their membrane disordering, and reduces the ability of aggregates to bend the membrane. Taken together, these phenomena may account for cholesterol’s effects on fengycin activity.