Systemic biodistribution and hepatocyte-specific gene editing with CRISPR/Cas9 using hyaluronic acid-based nanoparticles.

Autor: Francis C; Department of Pharmaceutical Sciences, School of Pharmacy, Northeastern University, Boston, MA, United States; Pfizer, Inc, Worldwide Research & Development, Medicinal Sciences, Biomedicine Design, Andover, MA, United States., Wroblewska L; Pfizer, Inc, Worldwide Research & Development, Medicinal Sciences, Biomedicine Design, Cambridge, MA, United States., Pegman P; Pfizer, Inc, Worldwide Research & Development, Medicinal Sciences, Bioprocess R&D, Andover, MA, United States., Amiji M; Department of Pharmaceutical Sciences, School of Pharmacy, Northeastern University, Boston, MA, United States. Electronic address: m.amiji@northeastern.edu.
Jazyk: angličtina
Zdroj: Nanomedicine : nanotechnology, biology, and medicine [Nanomedicine] 2022 Feb; Vol. 40, pp. 102488. Date of Electronic Publication: 2021 Nov 05.
DOI: 10.1016/j.nano.2021.102488
Abstrakt: The goal of this study was to evaluate hepatocyte-specific gene editing, via systemic administration of hyaluronic acid (HA)-based nanoparticles in naïve CD-1 mice. Using HA-poly(ethylene imine) (HA-PEI) and HA-PEI-mannose nanoparticles with differential mannose density (1X and 2X), we have evaluated systemic biodistribution and hepatocyte-specific delivery using IVIS imaging and flow cytometry. Additionally, we have investigated hepatocyte-specific delivery and transfection of CRISPR/Cas9 gene editing plasmid and eGFP gene payload to integrate at the Rosa26 locus. IVIS imaging showed uptake of HA-PEI nanoparticles primarily by the liver, and with addition of mannose at different concentrations, the nanoparticles showed increased uptake in both the liver and spleen. HA-PEI-mannose nanoparticles showed 55-65% uptake by hepatocytes, along with uptake by resident macrophage regardless of the mannose concentration. One of two gRNA targets showed 15% genome editing and obtained similar results for all three nanoparticle formulations. Cells positive for our gene payload were greatest with HA-PEI-mannose-1X nanoparticles where 16.2% of cells were GFP positive. The results were encouraging as proof of concept for the development of a non-viral biodegradable and biocompatible polymeric delivery system for gene editing specifically targeting hepatocytes upon systemic administration.
(Copyright © 2021 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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