2. Academic Validation
  3. A high-throughput Galectin-9 imaging assay for quantifying nanoparticle uptake, endosomal escape and functional RNA delivery

A high-throughput Galectin-9 imaging assay for quantifying nanoparticle uptake, endosomal escape and functional RNA delivery

  • Commun Biol. 2021 Feb 16;4(1):211. doi: 10.1038/s42003-021-01728-8.
Michael J Munson 1,  Gwen O'Driscoll 2,  Andreia M Silva 3,  Elisa Lázaro-Ibáñez 2,  Audrey Gallud 4,  John T Wilson 5,  Anna Collén 6,  Elin K Esbjörner 4,  Alan Sabirsh 7
Affiliations

Affiliations

  • 1 Advanced Drug Delivery, Pharmaceutical Sciences, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden. Michael.Munson@astrazeneca.com.
  • 2 Advanced Drug Delivery, Pharmaceutical Sciences, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden.
  • 3 Discovery Biology, Discovery Sciences, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden.
  • 4 Division of Chemical and Biomolecular Engineering, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden.
  • 5 Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN, USA.
  • 6 Projects, Research and Early Development, Cardiovascular, Renal and Metabolism, Biopharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden.
  • 7 Advanced Drug Delivery, Pharmaceutical Sciences, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden. Alan.Sabirsh@astrazeneca.com.
PMID: 33594247 DOI: 10.1038/s42003-021-01728-8
Abstract

RNA-based therapies have great potential to treat many undruggable human diseases. However, their efficacy, in particular for mRNA, remains hampered by poor cellular delivery and limited endosomal escape. Development and optimisation of delivery vectors, such as lipid nanoparticles (LNPs), are impeded by limited screening methods to probe the intracellular processing of LNPs in sufficient detail. We have developed a high-throughput imaging-based endosomal escape assay utilising a Galectin-9 reporter and fluorescently labelled mRNA to probe correlations between nanoparticle-mediated uptake, endosomal escape frequency, and mRNA translation. Furthermore, this assay has been integrated within a screening platform for optimisation of lipid nanoparticle formulations. We show that Galectin-9 recruitment is a robust, quantitative reporter of endosomal escape events induced by different mRNA delivery nanoparticles and small molecules. We identify nanoparticles with superior escape properties and demonstrate cell line variances in endosomal escape response, highlighting the need for fine-tuning of delivery formulations for specific applications.

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