Human and mouse long noncoding RNAs reengineered for exogenous delivery reduce LPS-induced inflammation in human macrophages and mice

Long noncoding RNAs (lncRNAs) constitute a substantial portion of the transcriptome and outnumber protein-coding transcripts in humans. lncRNA molecules are bioactive and control cellular and systemic functions by directly or indirectly regulating gene expression at the transcriptional or posttranscriptional levels. Here, we established the use of lncRNA as a modality to treat disease by reengineering three lncRNAs-GAPLINC, MIST, and DRAIR-to treat acute inflammation in mice and human macrophages. For each lncRNA, we established an in vitro transcription synthesis and high-performance liquid chromatography purification workflow and optimized the lncRNA isoforms, 5' cap, 3' poly(A) tail, and chemical base modifications to improve specificity and performance. We also optimized the in vivo delivery of lncRNA with a lipid nanoparticle system that did not induce confounding effects. Using this pipeline and delivery platform, we demonstrated that each lncRNA reduced lipopolysaccharide (LPS)-induced inflammation by specifically regulating distinct subsets of cytokines in cultured mouse macrophages and in mice. GAPLINC and DRAIR reduced the transcription of IL-1β and IL-6, respectively, whereas MIST attenuated TNFα production posttranscriptionally. In addition, we showed that reengineered GAPLINC reduced LPS-induced inflammation in human monocytes, suggesting the clinical potential of this approach. Our engineering approach and findings establish a previously unidentified nucleic acid modality and demonstrate an effective way to reengineer regulatory lncRNAs to treat disease.