Abstract
BackgroundRNA-based vaccination strategies tailoring immune response to specific reactions have become an important pillar for a broad range of applications. Recently, the use of lipid-based nanoparticles opened the possibility to deliver RNA to specific sites within the body, overcoming the limitation of rapid degradation in the bloodstream. Here, we have investigated whether small animal PET/MRI can be employed to image the biodistribution of RNA-encoded protein. For this purpose, a reporter RNA coding for the sodium-iodide-symporter (NIS) was in vitro transcribed in cell lines and evaluated for expression. RNA-lipoplex nanoparticles were then assembled by complexing RNA with liposomes at different charge ratios, and functional NIS protein translation was imaged and quantified in vivo and ex vivo by Iodine-124 PET upon intravenous administration in mice.ResultsNIS expression was detected on the membrane of two cell lines as early as 6 h after transfection and gradually decreased over 48 h. In vivo and ex vivo PET/MRI of anionic spleen-targeting or cationic lung-targeting NIS-RNA lipoplexes revealed a visually detectable rapid increase of Iodine-124 uptake in the spleen or lung compared to control-RNA-lipoplexes, respectively, with minimal background in other organs except from thyroid, stomach and salivary gland.ConclusionsThe strong organ selectivity and high target-to-background acquisition of NIS-RNA lipoplexes indicate the feasibility of small animal PET/MRI to quantify organ-specific delivery of RNA.
Highlights
RNA-based vaccination strategies tailoring immune response to specific reactions have become an important pillar for a broad range of applications
Due to the transient nature of RNA, the percentage of NIS-expressing cells decreased over time for both cell lines
Expression levels of NIS on the cell surface followed a similar pattern for both cell lines, peaking at 6 h after transfection (Fig. 1c, d)
Summary
RNA-based vaccination strategies tailoring immune response to specific reactions have become an important pillar for a broad range of applications. An optimized RNA-lipid composition with a net negative charge was shown to drive systemic RNA delivery to antigen-presenting cells, to spleen, and it has been translated into a delivery platform for cancer vaccination in clinical trials [2, 3, 6]. In contrast to those negatively charged RNA-lipoplexes, cationic RNA-lipoplexes showed similar size and stability but vastly different organ distribution with selective targeting of lung tissue [3, 7]. Targeting of immune cells of a certain organ— in this case the lung—is well possible with these systems
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