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Species-dependent in vivo mRNA delivery and cellular responses to nanoparticles

Abstract

Nanoparticles are tested in mice and non-human primates before being selected for clinical trials. Yet the extent to which mRNA delivery, as well as the cellular response to mRNA drug delivery vehicles, is conserved across species in vivo is unknown. Using a species-independent DNA barcoding system, we have compared how 89 lipid nanoparticles deliver mRNA in mice with humanized livers, primatized livers and four controls: mice with ‘murinized’ livers as well as wild-type BL/6, Balb/C and NZB/BlNJ mice. We assessed whether functional delivery results in murine, non-human primate and human hepatocytes can be used to predict delivery in the other species in vivo. By analysing in vivo hepatocytes by RNA sequencing, we identified species-dependent responses to lipid nanoparticles, including mRNA translation and endocytosis. These data support an evidence-based approach to making small-animal preclinical nanoparticle studies more predictive, thereby accelerating the development of RNA therapies.

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Fig. 1: Characterizing SANDS.
Fig. 2: Nanoparticle delivery across murine, NHP and human hepatocytes in vivo.
Fig. 3: Transcriptomic studies reveal species-dependent response to LNPs.
Fig. 4: Inflammatory genes impact mRNA delivery across multiple mouse strains.

Data availability

All RNA sequencing data have been deposited online at GEO (GSE178313). The scripts used to analyse barcodes are available at Github (https://github.com/Jack-Feldman/barcode_count). All other data are shown in the figures.

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Acknowledgements

The authors thank K. E. Tiegreen, S. Durham and R. Hughley at Georgia Institute of Technology. The work was funded by the National Institutes of Health (R01-GM132985, awarded to J.E.D., and UG3-TR002855, awarded to J.E.D. and P.J.S.) and DARPA (PREPARE, grant no. HR00111920008, awarded to P.J.S. and J.E.D.).

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M.Z.C.H., C.N.D., P.J.S. and J.E.D. conceived the experiments. P.J.S. and J.E.D. obtained funding for, and oversaw, the research. All authors performed the experiments. M.Z.C.H., C.N.D. and J.E.D. wrote the initial manuscript, which was edited by the authors.

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Correspondence to James E. Dahlman.

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J.E.D. is a consultant for GV. All other authors declare no competing interests.

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Nature Nanotechnology thanks David Morrissey and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Hatit, M.Z.C., Lokugamage, M.P., Dobrowolski, C.N. et al. Species-dependent in vivo mRNA delivery and cellular responses to nanoparticles. Nat. Nanotechnol. 17, 310–318 (2022). https://doi.org/10.1038/s41565-021-01030-y

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