Abstract
Therapeutic alteration of gene expression in vivo can be achieved by delivering nucleic acids (e.g., mRNA, siRNA) using nanoparticles. Recent progress in modified messenger RNA (mmRNA) synthesis facilitated the development of lipid nanoparticles (LNPs) loaded with mmRNA as a promising tool for in vivo protein expression. Although progress have been made with mmRNA-LNPs based protein expression in hepatocytes, cell specificity is still a major challenge. Moreover, selective protein expression is essential for an improved therapeutic effect, due to the heterogeneous nature of diseases. Here, we present a precision protein expression strategy in Ly6c+ inflammatory leukocytes in inflammatory bowel disease (IBD) induced mice. We demonstrate a therapeutic effect in an IBD model by targeted expression of the interleukin 10 in Ly6c+ inflammatory leukocytes. A selective mmRNA expression strategy has tremendous therapeutic potential in IBD and can ultimately become a novel therapeutic modality in many other diseases.
Highlights
Therapeutic alteration of gene expression in vivo can be achieved by delivering nucleic acids using nanoparticles
The formulation of lipid nanoparticles (LNPs) was designed and optimized to enable a controllable modified messenger RNA (mmRNA) encapsulation and to promote an efficient gene expression in leukocytes. mmRNA was formulated in LNPs using the NanoAssemblr® microfluidic mixing system (Precision Nanosystems Inc., Vancouver, Canada), in which mRNA molecules self-assembled with ionizable lipids in acidic condition to form highly uniform nanoparticles, of 63.7 ± 1.59 nm in diameter
We have evaluated the ability of mmRNA LNPs to promote protein expression in vitro in Raw 264.7 macrophages cell line (Supplementary Figure 1c−d) and ex-vivo in mouse splenocytes
Summary
Therapeutic alteration of gene expression in vivo can be achieved by delivering nucleic acids (e.g., mRNA, siRNA) using nanoparticles. Recent insights into mRNA structure and function, together with the advances in in vitro transcription methods and the introduction of modified nucleotides (e.g., 5mC, pseudo-Uridine etc.), facilitate the utilization of mRNA for therapeutic applications with higher expression efficiencies and lower immunogenicity[1,2,3] Both strategies to manipulate protein expression require the untrivial intracellular delivery of RNA molecules. A critical challenge of RNA-based therapeutic approach lies in the ability to deliver RNA molecules effectively to specific target cells These challenges promoted the development of synthetic and natural delivery systems as a promising strategy for non-viral and viral gene manipulation, respectively. Another advantage of mRNA-based therapies is the lower risk for genomic integration, which is a major risk factor in DNA based treatments
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