single cell sequencing of mouse lung single cell suspensions
Dysfunction of endothelial cells lining the inner surface of blood vessels are causative for a number of diseases. Hence, the ability to therapeutically modulate gene expression within endothelial cells is of high therapeutic value in treating diseases such as those associated with lung edema. mRNAs formulated with lipid nanoparticles (LNPs) have emerged as a new drug modality to induce transient protein expression for modulating disease relevant signal transduction pathways. In the study presented here we tested the effect of a novel synthetic, nucleoside-modified mRNA encoding COMP-Ang1 (mRNA-76) formulated into a cationic lipid nanoparticle (cLNP) on attenuating inflammation-induced vascular leakage. After intravenous injection, the respective mRNA was found to be delivered almost exclusively to the endothelial cells of the lung, while sparing other vascular beds and bypassing the liver. The mode of action of mRNA-76, such as its activation of the Tie2 signal transduction pathway, was tested by pharmacological studies in vitro and in vivo in respective mouse models. mRNA-76 was found to prevent lung vascular leakage/lung edema as well as neutrophil infiltration in an LPS-challenging model. Overall design: To investigate the specific cellular uptake of two different mRNAs in mice lung following intravenous delivery of encapsulated mRNA in the same lipid nanoparticle delivery system following 15 and 24 hours of administration. Gene expression profiling was performed to identify lung cell types and the two target mRNAs: one encoding a potent tie2 agonist COMP-ANG1 (mRNA-76; PAN004) and the other control mRNA encoding for a non-secreted firefly luciferase (mRNAFLUC; PAN93). Comparative gene expression profiling anlaysis of RNA-seq data for mouse lung cells, mRNA-76 and mRNAFLUC.