Species-agnostic polymeric formulations for inhalable messenger RNA delivery to the lung

doi:10.1038/s41563-022-01404-0

. 2022 Nov 28.

doi: 10.1038/s41563-022-01404-0. Online ahead of print.

Species-agnostic polymeric formulations for inhalable messenger RNA delivery to the lung

Laura Rotolo^#¹, Daryll Vanover^#¹, Nicholas C Bruno^#², Hannah E Peck¹, Chiara Zurla¹, Jackelyn Murray³, Richard K Noel⁴, Laura O'Farrell⁴, Mariluz Araínga⁵, Nichole Orr-Burks³, Jae Yeon Joo¹, Lorena C S Chaves¹, Younghun Jung¹, Jared Beyersdorf¹, Sanjeev Gumber⁶, Ricardo Guerrero-Ferreira⁷, Santiago Cornejo⁸, Merrilee Thoresen⁸, Alicia K Olivier⁸, Katie M Kuo², James C Gumbart^{2

9}, Amelia R Woolums⁸, Francois Villinger⁵, Eric R Lafontaine³, Robert J Hogan^{3

10}, M G Finn^{2

11}, Philip J Santangelo¹²

Affiliations

¹ Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
² School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA.
³ Department of Infectious Diseases, College of Veterinary Medicine University of Georgia, Athens, GA, USA.
⁴ Physiological Research Laboratory, Georgia Institute of Technology, Atlanta, GA, USA.
⁵ New Iberia Research Center, University of Louisiana Lafayette, Lafayette, LA, USA.
⁶ Yerkes National Primate Research Center, Emory University, Atlanta, GA, USA.
⁷ Robert P. Apkarian Integrated Electron Microscopy Core, Emory University, Atlanta, GA, USA.
⁸ Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS, USA.
⁹ School of Physics, Georgia Institute of Technology, Atlanta, GA, USA.
¹⁰ Department of Veterinary Biosciences and Diagnostic Imaging, College of Veterinary Medicine University of Georgia, Athens, GA, USA.
¹¹ School of Biological Sciences, Georgia Institute of Technology, Atlanta, GA, USA.
¹² Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA. philip.j.santangelo@emory.edu.

^# Contributed equally.

PMID: 36443576
DOI: 10.1038/s41563-022-01404-0

Species-agnostic polymeric formulations for inhalable messenger RNA delivery to the lung

Laura Rotolo et al. Nat Mater. 2022.

. 2022 Nov 28.

doi: 10.1038/s41563-022-01404-0. Online ahead of print.

Authors

Affiliations

¹ Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
² School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA.
³ Department of Infectious Diseases, College of Veterinary Medicine University of Georgia, Athens, GA, USA.
⁴ Physiological Research Laboratory, Georgia Institute of Technology, Atlanta, GA, USA.
⁵ New Iberia Research Center, University of Louisiana Lafayette, Lafayette, LA, USA.
⁶ Yerkes National Primate Research Center, Emory University, Atlanta, GA, USA.
⁷ Robert P. Apkarian Integrated Electron Microscopy Core, Emory University, Atlanta, GA, USA.
⁸ Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS, USA.
⁹ School of Physics, Georgia Institute of Technology, Atlanta, GA, USA.
¹⁰ Department of Veterinary Biosciences and Diagnostic Imaging, College of Veterinary Medicine University of Georgia, Athens, GA, USA.
¹¹ School of Biological Sciences, Georgia Institute of Technology, Atlanta, GA, USA.
¹² Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA. philip.j.santangelo@emory.edu.

^# Contributed equally.

PMID: 36443576
DOI: 10.1038/s41563-022-01404-0

Abstract

Messenger RNA has now been used to vaccinate millions of people. However, the diversity of pulmonary pathologies, including infections, genetic disorders, asthma and others, reveals the lung as an important organ to directly target for future RNA therapeutics and preventatives. Here we report the screening of 166 polymeric nanoparticle formulations for functional delivery to the lungs, obtained from a combinatorial synthesis approach combined with a low-dead-volume nose-only inhalation system for mice. We identify P76, a poly-β-amino-thio-ester polymer, that exhibits increased expression over formulations lacking the thiol component, delivery to different animal species with varying RNA cargos and low toxicity. P76 allows for dose sparing when delivering an mRNA-expressed Cas13a-mediated treatment in a SARS-CoV-2 challenge model, resulting in similar efficacy to a 20-fold higher dose of a neutralizing antibody. Overall, the combinatorial synthesis approach allowed for the discovery of promising polymeric formulations for future RNA pharmaceutical development for the lungs.

References

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Grant support

HR00111920008/United States Department of Defense | Defense Advanced Research Projects Agency (DARPA)

[1] Pardi, N. et al. Expression kinetics of nucleoside-modified mRNA delivered in lipid nanoparticles to mice by various routes. J. Control. Release 217, 345–351 (2015). - DOI

[2] Pardi, N. et al. Expression kinetics of nucleoside-modified mRNA delivered in lipid nanoparticles to mice by various routes. J. Control. Release 217, 345–351 (2015). - DOI

[3] Tiwari, P. M. et al. Engineered mRNA-expressed antibodies prevent respiratory syncytial virus infection. Nat. Commun. 9, 3999 (2018). - DOI

[4] Tiwari, P. M. et al. Engineered mRNA-expressed antibodies prevent respiratory syncytial virus infection. Nat. Commun. 9, 3999 (2018). - DOI

[5] Qiu, Y. et al. Effective mRNA pulmonary delivery by dry powder formulation of PEGylated synthetic KL4 peptide. J. Control. Release 314, 102–115 (2019). - DOI

[6] Qiu, Y. et al. Effective mRNA pulmonary delivery by dry powder formulation of PEGylated synthetic KL4 peptide. J. Control. Release 314, 102–115 (2019). - DOI

[7] Lokugamage, M. P. et al. Optimization of lipid nanoparticles for the delivery of nebulized therapeutic mRNA to the lungs. Nat. Biomed. Eng. 5, 1059–1068 (2021). - DOI

[8] Lokugamage, M. P. et al. Optimization of lipid nanoparticles for the delivery of nebulized therapeutic mRNA to the lungs. Nat. Biomed. Eng. 5, 1059–1068 (2021). - DOI

[9] Chollet, P., Favrot, M. C., Hurbin, A. & Coll, J.-L. Side-effects of a systemic injection of linear polyethylenimine–DNA complexes. J. Gene Med. 4, 84–91 (2002). - DOI

[10] Chollet, P., Favrot, M. C., Hurbin, A. & Coll, J.-L. Side-effects of a systemic injection of linear polyethylenimine–DNA complexes. J. Gene Med. 4, 84–91 (2002). - DOI