Abstract
Introduction
Lipid nanoparticles (LNPs) tend to accumulate in the liver due to physiological factors. Whereas the biological mechanisms that promote LNP delivery to hepatocytes have been reported, the mechanisms that promote delivery to other cell types within the liver microenvironment are poorly understood. Single cell profiling studies have recently identified subsets of Kupffer cells and hepatic endothelial cells with distinct gene expression patterns and biological phenotypes; we hypothesized these subtypes would differentially interact with nanoparticles.
Methods
To test the hypothesis, we quantified nucleic acid (i) biodistribution and (ii) functional mRNA delivery within the liver microenvironment using two clinically relevant LNPs in vivo.
Results
We found that these LNPs distribute nucleic acids distribute to Kupffer cells and liver endothelial cells as efficiently as they distribute to hepatocytes, yet result in more functional mRNA delivery to endothelial cells. Additionally, we found these LNPs differentially accumulate in Kupffer and endothelial cell subsets.
Conclusions
These data suggest subsets of liver microenvironmental cells can differentially interact with nanoparticles in vivo, thereby altering LNP delivery. More generally, the data suggest that nucleic acid biodistribution is not sufficient to predict functional nucleic acid delivery in vivo.
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Acknowledgments
The authors thank Sommer Durham and the Georgia Tech Cellular Analysis and Cytometry Core. Additionally, the authors thank Dalia Arafat and the Genome Analysis Core. J.E.D. thanks Jordan Cattie and Taylor E. Shaw.
Conflict of interest
Cory D. Sago is co-founder of Guide Therapeutics and an employee at Guide Therapeutics. James E. Dahlman is a co-founder of Guide Therapeutics and consultant for Guide Therapeutics. Brandon R. Krupczak is an employee at Guide Therapeutics. Melissa P. Lokugamage declares no conflict of interest. Zubao Gan declares no conflict of interest.
Ethical Approval
All animal studies were carried out in accordance with the institutional and national guidelines, following an animal protocol approved by the Georgia Institute of Technology IACUC committee. No human studies were performed as part of this research.
Funding
C.D.S. and J.E.D. were funded by Georgia Tech startup funds (awarded to J.E.D.). C.D.S. was funded by the NIH-sponsored Research Training Program in Immunoengineering (T32EB021962). M.P.L was funded by the NIH-sponsored Research Training Program in Computational Biology and Predictive Health Genomics (T32GM105490). This content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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C.D.S. and J.E.D. designed experiments, performed experiments, and analyzed data. B.Z.K., M.P.L., Z.G. performed experiments. C.D.S. and J.E.D. wrote the paper, which was reviewed by all other authors.
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Associate Editor Michael R. King oversaw the review of this article.
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James E. Dahlman is an Assistant Professor in the Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory. His lab works at the interface of drug delivery, nanotechnology, genomics, and gene editing. James has designed nanoparticles that deliver RNA to blood vessels in the heart and lung; these nanoparticles have been validated by > 20 labs and have been licensed for clinical development. James also uses molecular biology to design the genetic drugs he delivers. He designed ‘dead’ guide RNAs to turn on genes using active Cas9. Similarly, using his background in nanoparticle chemistry, in vivo RNA delivery, and genomics, his lab has designed a series of increasingly sensitive DNA barcoding systems that can measure how > 200 nanoparticles target cells 30 different cell types at once, directly in vivo. James’ nanoparticle barcoding work led to his placement on the MIT Tech Review TR35 list. James has won scientific awards at every stage of his career, including the NSF, NDSEG, NIH OxCam, Whitaker, and LSRF Fellowships, and the Weintraub Graduate Thesis Award. He has been named a young / leading investigator by Bayer, the Parkinson’s Disease Foundation, and the Journal of Materials Chemistry B. At the age of 32, his research has been published in Science, Cell, Nature Nanotechnology, Nature Biotechnology, Nature Cell Bio, Science Translational Medicine, PNAS, Advanced Materials, JACS and other leading journals. He has given > 75 invited talks on drug delivery, gene editing, and nanoparticle DNA barcoding across the world, and is a co-founder of GuideRx.
This article is part of the 2019 CMBE Young Innovators special issue.
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Sago, C.D., Krupczak, B.R., Lokugamage, M.P. et al. Cell Subtypes Within the Liver Microenvironment Differentially Interact with Lipid Nanoparticles. Cel. Mol. Bioeng. 12, 389–397 (2019). https://doi.org/10.1007/s12195-019-00573-4
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DOI: https://doi.org/10.1007/s12195-019-00573-4