Optimization-by-design of hepatotropic lipid nanoparticles targeting the sodium-taurocholate cotransporting polypeptide
- University of Basel, Switzerland
- University Hospital Heidelberg, Germany
Abstract
Active targeting and specific drug delivery to parenchymal liver cells is a promising strategy to treat various liver disorders. Here, we modified synthetic lipid-based nanoparticles with targeting peptides derived from the hepatitis B virus large envelope protein (HBVpreS) to specifically target the sodium-taurocholate cotransporting polypeptide (NTCP; SLC10A1) on the sinusoidal membrane of hepatocytes. Physicochemical properties of targeted nanoparticles were optimized and NTCP-specific, ligand-dependent binding and internalization was confirmed in vitro. The pharmacokinetics and targeting capacity of selected lead formulations was investigated in vivo using the emerging zebrafish screening model. Liposomal nanoparticles modified with 0.25 mol% of a short myristoylated HBV derived peptide, i.e. Myr‑HBVpreS2-31, showed an optimal balance between systemic circulation, avoidance of blood clearance, and targeting capacity. Pronounced liver enrichment, active NTCP‑mediated targeting of hepatocytes and efficient cellular internalization were confirmed in mice by 111In gamma scintigraphy and fluorescence microscopy demonstrating the potential use of our hepatotropic, ligand-modified nanoparticles.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files. Numerical data for all quantitative graphs are provided in the Figure source data.
Article and author information
Author details
Henriette E Meyer zu Schwabedissen
Funding
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (174975)
- Dominik Witzigmann
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (173057)
- Jonas Buck
- Jörg Huwyler
Deutsche Forschungsgemeinschaft (209091148)
- Stephan Urban
Deutsches Zentrum für Infektionsforschung (5.704)
- Stephan Urban
Deutsches Zentrum für Infektionsforschung (5.807)
- Stephan Urban
Freiwillige Akademische Gesellschaft (FAG Basel)
- Dominik Witzigmann
- Sandro Sieber
- Jörg Huwyler
Stiftung zur Förderung des pharmazeutischen Nachwuchses in Basel
- Sandro Sieber
Novartis University Basel Excellence SCholarship for Life Sciences
- Dominik Witzigmann
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: Animal experimentation: Zebrafish embryo (Danio rerio) studies were performed in strict accordance with Swiss animal welfare regulations. Mouse and rat experiments were carried out in accordance with German legislation on animal welfare. All of the animals were handled according to approved institutional animal care and use protocol of the University of Basel and University of Heidelberg.
Copyright
© 2019, Witzigmann et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Optimization-by-design of hepatotropic lipid nanoparticles targeting the sodium-taurocholate cotransporting polypeptide
eLife 8:e42276.
https://doi.org/10.7554/eLife.42276
Further reading
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- Biochemistry and Chemical Biology
- Microbiology and Infectious Disease
Teichoic acids (TA) are linear phospho-saccharidic polymers and important constituents of the cell envelope of Gram-positive bacteria, either bound to the peptidoglycan as wall teichoic acids (WTA) or to the membrane as lipoteichoic acids (LTA). The composition of TA varies greatly but the presence of both WTA and LTA is highly conserved, hinting at an underlying fundamental function that is distinct from their specific roles in diverse organisms. We report the observation of a periplasmic space in Streptococcus pneumoniae by cryo-electron microscopy of vitreous sections. The thickness and appearance of this region change upon deletion of genes involved in the attachment of TA, supporting their role in the maintenance of a periplasmic space in Gram-positive bacteria as a possible universal function. Consequences of these mutations were further examined by super-resolved microscopy, following metabolic labeling and fluorophore coupling by click chemistry. This novel labeling method also enabled in-gel analysis of cell fractions. With this approach, we were able to titrate the actual amount of TA per cell and to determine the ratio of WTA to LTA. In addition, we followed the change of TA length during growth phases, and discovered that a mutant devoid of LTA accumulates the membrane-bound polymerized TA precursor.
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- Biochemistry and Chemical Biology
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