Enabling non-viral DNA delivery using lipid nanoparticles co-loaded with endogenous anti-inflammatory lipids

  • bioRxiv. 2024 Jun 12:2024.06.11.598533. doi: 10.1101/2024.06.11.598533.
Manthan N Patel  1  2 ,  Sachchidanand Tiwari  2  3 ,  Yufei Wang  2 ,  Sarah O'Neill  1 ,  Jichuan Wu  2 ,  Serena Omo-Lamai  2  4 ,  Carolann Espy  1  2 ,  Liam S Chase  2  4 ,  Aparajeeta Majumdar  2 ,  Evan Hoffman  2 ,  Anit Shah  2 ,  András Sárközy  3 ,  Jeremy Katzen  2 ,  Norbert Pardi  3 ,  Jacob S Brenner  1  2
Affiliations
  • 1. Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
  • 2. Division of Pulmonary, Allergy, and Critical Care, Department of Medicine, Perelman School of Medicine University of Pennsylvania Philadelphia, PA, USA.
  • 3. Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
  • 4. Department of Bioengineering, School of Engineering & Applied Science, University of Pennsylvania, Philadelphia, PA, USA.
PMID: 38915627 DOI: 10.1101/2024.06.11.598533
Abstract

Lipid nanoparticles (LNPs) have transformed genetic medicine, recently shown by their use in COVID-19 mRNA vaccines. While loading LNPs with mRNA has many uses, loading DNA would provide additional advantages such as long-term expression and availability of promoter sequences. However, here we show that plasmid DNA (pDNA) delivery via LNPs (pDNA-LNPs) induces acute inflammation in naïve mice which we find is primarily driven by the cGAS-STING pathway. Inspired by DNA viruses that inhibit this pathway for replication, we co-loaded endogenous lipids that inhibit STING into pDNA-LNPs. Specifically, loading nitro-oleic acid (NOA) into pDNA-LNPs (NOA-pDNA-LNPs) ameliorates serious inflammatory responses in vivo enabling prolonged transgene expression (at least 1 month). Additionally, we demonstrate the ability to iteratively optimize NOA-pDNA-LNPs' expression by performing a small LNP formulation screen, driving up expression 50-fold in vitro. Thus, NOA-pDNA-LNPs, and pDNA-LNPs co-loaded with Other bioactive molecules, will provide a major new tool in the genetic medicine toolbox, leveraging the power of DNA's long-term and promoter-controlled expression.

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