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Combinatorial design of nanoparticles for pulmonary mRNA delivery and genome editing

Nature Biotechnology (2023)Cite this article

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Abstract

The expanding applications of nonviral genomic medicines in the lung remain restricted by delivery challenges. Here, leveraging a high-throughput platform, we synthesize and screen a combinatorial library of biodegradable ionizable lipids to build inhalable delivery vehicles for messenger RNA and CRISPR–Cas9 gene editors. Lead lipid nanoparticles are amenable for repeated intratracheal dosing and could achieve efficient gene editing in lung epithelium, providing avenues for gene therapy of congenital lung diseases.

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Fig. 1: Overview of LNP synthesis and screening.
Fig. 2: Gene editing in mouse lung with RCB-4-8 LNP-mRNA.

Data availability

Illumina Sequencing data have been submitted to the Sequence Read Archive; these datasets are available under BioProject Accession no. PRJNA918691 (ref. 37). The authors declare that all other data supporting the findings of this study are available within the paper and its Supplementary Information files or on reasonable request. Plasmids are available from Addgene.

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Acknowledgements

This work was supported by Translate Bio and the National Institutes of Health (grant no. UG3HL147367). B.L., R.S.M., A.G. and A.J. were supported by Translate Bio. B.L. was supported by the Leslie Dan Faculty of Pharmacy startup fund, the Connaught Fund (no. 514681), the J. P. Bickell Foundation (grant no. 515159), the Canada Research Chairs Program (no. CRC-2022-00575), Canadian Institutes of Health Research (no. PJH-185722) and the Canada Foundation for Innovation John R. Evans Leaders Fund (no. 43711). A.V. was supported by the PRiME Postdoctoral Fellowship from the University of Toronto. S.P.L., G.G., W.X. and D.A. were supported by grants from the National Institutes of Health (nos. UG3HL147367 and UH3HL147367). W.X. was supported by grants from the National Institutes of Health (nos. DP2HL137167 and P01HL131471), the American Cancer Society (grant no. 129056-RSG-16-093) and the Cystic Fibrosis Foundation. We thank the Koch Institute Swanson Biotechnology Center for technical support, specifically the Animal Imaging & Preclinical Testing, Histology, Nanotechnology Materials and Microscopy core facilities.

Author information

Author notes

  1. These authors contributed equally: Bowen Li, Rajith Singh Manan, Shun-Qing Liang.

Authors and Affiliations

  1. David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA

    Bowen Li, Rajith Singh Manan, Akiva Gordon, Allen Jiang, Robert Langer & Daniel Anderson

  2. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    Bowen Li, Rajith Singh Manan, Akiva Gordon, Allen Jiang, Robert Langer & Daniel Anderson

  3. Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario, Canada

    Bowen Li & Andrew Varley

  4. Institute of Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada

    Bowen Li

  5. RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA

    Shun-Qing Liang, Guangping Gao & Wen Xue

  6. Horae Gene Therapy Center and Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA, USA

    Guangping Gao

  7. Department of Anesthesiology, Boston Children’s Hospital, Boston, MA, USA

    Robert Langer & Daniel Anderson

  8. Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA, USA

    Robert Langer & Daniel Anderson

  9. Harvard–MIT Division of Health Science and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA

    Robert Langer & Daniel Anderson

  10. Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, MA, USA

    Wen Xue

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Contributions

B.L., R.S.M. and S.Q.L. conceived the project and wrote the paper, with input from all authors. B.L. and R.S.M. designed the combinatorial lipid library. B.L., R.S.M., S.Q.L., A.G. and A.J. performed experiments and analyzed data. B.L., R.S.M. and S.Q.L. wrote the paper. B.L., S.Q.L., A.V., W.X. and D.G.A. discussed the results and edited the paper. G.G., R.L., W.X. and D.A. acquired funding and supervised the project.

Corresponding authors

Correspondence to Wen Xue or Daniel Anderson.

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Competing interests

B.L., R.S.M., A.G. and D.A. have filed a patent (PCT/US2022/052314) for development of the described lipids. D.A. receives research funding from Translate Bio and is a Founder of Orna Therapeutics. R.L. is a cofounder of Moderna; he also serves on the board and has equity in Particles for Humanity. For a list of entities with which R.L. is, or has been, recently involved, compensated or uncompensated, see https://www.dropbox.com/s/yc3xqb5s8s94v7x/Rev%20Langer%20COI.pdf?dl=0. The other authors declare no competing interests.

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Li, B., Manan, R.S., Liang, SQ. et al. Combinatorial design of nanoparticles for pulmonary mRNA delivery and genome editing. Nat Biotechnol (2023). https://doi.org/10.1038/s41587-023-01679-x

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