Total RNA expression in extracellular vesicles (EVs) derived from untreated and lipid nanoparticle(LNP) treated cells: lung epithelial, umbilical vein endothelial, and cardiac progenitor cells

The study used next-generation sequencing to compare mRNA content of EVs from 3 different cell origins which are untreated control or treated with LNP containing VEGFA mRNA. Our goal is to carry out functional and pathway analysis of the differentially expressed genes. The RNA profiles of EV content from treated and untreated cells; lung epithelial (HTB-177) , umbilical vein endothelial (HUVEC), and cardiac progenitor cells(CPC), were generated by RNA-Seq using Illumina NovaSeq6000. Differentially expressed genes were identified using the DESeq2 package. We found that the VEGFA mRNA was among the most upregulated genes in EVs from LNP treated cells. We also identified multiple genes and pathways associated with angiogenesis and cardiac conditions that were activated in the EVs derived from treated cells. The result suggests that VEGFA-LNP were processed by the cells and certain portions of the mRNA were packaged into EVs and released into the cell culture medium. Overall design: mRNA profiles of EVs derived from HTB-177 (n = 4) , HUVEC (n = 4) or CPC (n = 3) cells which were untreated or treated with LNP containing VEGFA mRNA