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Optimization of lipid nanoparticles for the delivery of nebulized therapeutic mRNA to the lungs

Nature Biomedical Engineering volume 5, pages 1059–1068 (2021)Cite this article

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Abstract

Lipid nanoparticles (LNPs) for the efficient delivery of drugs need to be designed for the particular administration route and type of drug. Here we report the design of LNPs for the efficient delivery of therapeutic RNAs to the lung via nebulization. We optimized the composition, molar ratios and structure of LNPs made of lipids, neutral or cationic helper lipids and poly(ethylene glycol) (PEG) by evaluating the performance of LNPs belonging to six clusters occupying extremes in chemical space, and then pooling the lead clusters and expanding their diversity. We found that a low (high) molar ratio of PEG improves the performance of LNPs with neutral (cationic) helper lipids, an identified and optimal LNP for low-dose messenger RNA delivery. Nebulized delivery of an mRNA encoding a broadly neutralizing antibody targeting haemagglutinin via the optimized LNP protected mice from a lethal challenge of the H1N1 subtype of influenza A virus, and delivered mRNA more efficiently than LNPs previously optimized for systemic delivery. A cluster approach to LNP design may facilitate the optimization of LNPs for other administration routes and therapeutics.

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Fig. 1: An in vivo workflow to evaluate how chemically diverse LNPs deliver mRNA to the lung after nebulization.
Fig. 2: Surveying how four LNP chemical traits influence nebulized lung mRNA delivery in vivo.
Fig. 3: A screen of LNPs containing cationic lipids from an expanded chemical space.
Fig. 4: A screen of LNPs containing neutral lipids from an expanded chemical space.
Fig. 5: Design rules and optimized LNPs for therapeutic nebulized mRNA delivery.

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Data availability

The main data supporting the results in this study are available within the paper and its Supplementary Information. Data on the expression of inflammatory genes are available as Supplementary Information. The raw and analyzed datasets generated during the study are too large to be publicly shared, but they are available for research purposes from the corresponding authors on reasonable request.

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Acknowledgements

We thank the Emory Integrated Genomics Core (EIGC) at Emory University and the Emory Winship Cancer Institute Cancer Tissue and Pathology Core (CTP). J.E.D. thanks T. E. Shaw. The work was funded by Emily’s Entourage (awarded to J.E.D.), the National Institutes of Health (UG3-TR002855, awarded to J.E.D. and P.J.S.) and DARPA (PREPARE grant number HR00111920008, awarded to P.J.S. and J.E.D.).

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Author notes

  1. These authors contributed equally: Melissa P. Lokugamage, Daryll Vanover, Jared Beyersdorf, Marine Z. C. Hatit.

Authors and Affiliations

  1. Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA, USA

    Melissa P. Lokugamage, Daryll Vanover, Jared Beyersdorf, Marine Z. C. Hatit, Laura Rotolo, Elisa Schrader Echeverri, Hannah E. Peck, Huanzhen Ni, YongTae Kim, Philip J. Santangelo & James E. Dahlman

  2. George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA, USA

    Jeong-Kee Yoon & YongTae Kim

  3. Parker H. Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, GA, USA

    YongTae Kim, Philip J. Santangelo & James E. Dahlman

  4. Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, GA, USA

    YongTae Kim

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Contributions

M.P.L., D.V., J.B., P.J.S. and J.E.D. conceived the experiments. M.P.L., D.V., J.B., Y.K., P.J.S. and J.E.D. designed the experiments. M.P.L., D.V., J.B., M.Z.C.H., L.R., E.S.E., H.E.P., H.N. and J.-K.Y. performed the experiments. M.P.L. and J.E.D. wrote the initial draft, which was edited by all authors. P.J.S. and J.E.D. oversaw the research.

Corresponding authors

Correspondence to Philip J. Santangelo or James E. Dahlman.

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Competing interests

J.E.D. is a consultant for Beam Therapeutics and GV. The other authors declare no competing interests.

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Peer review information Nature Biomedical Engineering thanks Dan Peer, Holun Wong and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary Table 1

Expression of inflammatory genes.

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Lokugamage, M.P., Vanover, D., Beyersdorf, J. et al. Optimization of lipid nanoparticles for the delivery of nebulized therapeutic mRNA to the lungs. Nat Biomed Eng 5, 1059–1068 (2021). https://doi.org/10.1038/s41551-021-00786-x

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