Cationic cholesterol-dependent LNP delivery to lung stem cells, the liver, and heart

Abstract

Adding a cationic helper lipid to a lipid nanoparticle (LNP) can increase lung delivery and decrease liver delivery. However, it remains unclear whether charge-dependent tropism is universal or, alternatively, whether it depends on the component that is charged. Here, we report evidence that cationic cholesterol-dependent tropism can differ from cationic helper lipid-dependent tropism. By testing how 196 LNPs delivered mRNA to 22 cell types, we found that charged cholesterols led to a different lung:liver delivery ratio than charged helper lipids. We also found that combining cationic cholesterol with a cationic helper lipid led to mRNA delivery in the heart as well as several lung cell types, including stem cell-like populations. These data highlight the utility of exploring charge-dependent LNP tropism.

Organ- and Cell-Selective Delivery of mRNA In Vivo Using Guanidinylated Serinol Charge-Altering Releasable Transporters

“…Further, when incubated in mouse plasma, these nanoparticles showed a more significant increase in size putatively due to their association with proteins. The strategy of tuning charge ratio shares partial similarity with previous work on organ-tropic LNP systems, 13,14 in which the addition of cationic lipids promoted lung tropism while anionic lipids promoted spleen tropism. 16 However, neither the size nor the surface charge has been reported to be correlated with organ tropism in LNP systems, 14 suggesting that organ-targeting mechanisms for LNPs and CARTs could be different.…”

“…12 More recently, and somewhat unexpectedly, "passive targeting", based on the intrinsic properties of LNP−mRNA complexes, has shown considerable promise for organ-selective delivery. 13,14 For instance, the Sahin group has shown that liposome formulations with varied charge ratios (cationic lipid to anionic mRNA phosphates), upon i.v. administration, selectively transfect lungs and spleens.…”

“…“Corona effects”, in which LNP surfaces are modified postadministration through association with ligands in the blood, have also effectively participated in organ-selective delivery (e.g., APOE for liver delivery) . More recently, and somewhat unexpectedly, “passive targeting”, based on the intrinsic properties of LNP–mRNA complexes, has shown considerable promise for organ-selective delivery. , For instance, the Sahin group has shown that liposome formulations with varied charge ratios (cationic lipid to anionic mRNA phosphates), upon i.v. administration, selectively transfect lungs and spleens .…”