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Nanoparticle single-cell multiomic readouts reveal that cell heterogeneity influences lipid nanoparticle-mediated messenger RNA delivery

Nature Nanotechnology volume 17, pages 871–879 (2022)Cite this article

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Abstract

Cells that were previously described as homogeneous are composed of subsets with distinct transcriptional states. However, it remains unclear whether this cell heterogeneity influences the efficiency with which lipid nanoparticles (LNPs) deliver messenger RNA therapies in vivo. To test the hypothesis that cell heterogeneity influences LNP-mediated mRNA delivery, we report here a new multiomic nanoparticle delivery system called single-cell nanoparticle targeting-sequencing (SENT-seq). SENT-seq quantifies how dozens of LNPs deliver DNA barcodes and mRNA into cells, the subsequent protein production and the transcriptome, with single-cell resolution. Using SENT-seq, we have identified cell subtypes that exhibit particularly high or low LNP uptake as well as genes associated with those subtypes. The data suggest that cell subsets have distinct responses to LNPs that may affect mRNA therapies.

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Fig. 1: SENT-seq uses orthogonal capture sequences to generate tunable multiomic readouts.
Fig. 2: In vivo multiomic single-cell readouts of the transcriptome, functional LNP-mediated mRNA delivery and LNP-mediated DNA barcode delivery.
Fig. 3: Cell subsets differentially uptake LNPs.
Fig. 4: Endothelial cell subtypes show transcriptional differences that may dictate LNP-mediated mRNA delivery.
Fig. 5: Chemically distinct LNPs exhibit different tropism within the liver microenvironment.

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Data availability

All raw sequencing data (GEO: GSE186395) are available online. All other data are provided in the main text or the Supplementary Information.

Code availability

The custom code used is available at https://github.com/Jack-Feldman/barcode_count.

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Acknowledgements

We thank K. Tiegren for copyediting the manuscript. Funding was provided by the National Institutes of Health (grant nos. UG3-TR002855 (J.E.D. and P.J.S.) and R01DE026941 (J.E.D.)).

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Author notes

  1. These authors contributed equally: Curtis Dobrowolski, Kalina Paunovska.

Authors and Affiliations

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

    Curtis Dobrowolski, Kalina Paunovska, Elisa Schrader Echeverri, David Loughrey, Huanzhen Ni, Marine Z. C. Hatit, Melissa P. Lokugamage, Hannah E. Peck, Philip J. Santangelo & James E. Dahlman

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

    Alejandro J. Da Silva Sanchez & Yanina Kuzminich

  3. Department of Chemical Engineering, Georgia Institute of Technology, Atlanta, GA, USA

    Alejandro J. Da Silva Sanchez

  4. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA, USA

    Yanina Kuzminich

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Contributions

Conceptualization: C.D., K.P. and J.E.D. Methodology: C.D., K.P. and J.E.D. Investigation: C.D., K.P., E.S.E., D.L., A.J.D.S.S., H.N., M.Z.C.H., M.P.L., Y.K., H.E.P., P.J.S. and J.E.D. Visualization: C.D., K.P. and J.E.D. Funding acquisition: J.E.D. and P.J.S. Project administration: J.E.D. Supervision: P.J.S. and J.E.D. Writing—original draft: C.D., K.P. and J.E.D. Writing—review and editing: C.D., K.P., E.S.E., D.L., A.J.D.S.S., H.N., M.Z.C.H., M.P.L., Y.K., H.E.P., P.J.S. and J.E.D.

Corresponding author

Correspondence to James E. Dahlman.

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Competing interests

C.D., K.P. and J.E.D. have filed intellectual property related to SENT-seq. J.E.D. is an advisor for GV. All other authors declare no competing interests.

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Dobrowolski, C., Paunovska, K., Schrader Echeverri, E. et al. Nanoparticle single-cell multiomic readouts reveal that cell heterogeneity influences lipid nanoparticle-mediated messenger RNA delivery. Nat. Nanotechnol. 17, 871–879 (2022). https://doi.org/10.1038/s41565-022-01146-9

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