PEGylation of recombinant human deoxyribonuclease I decreases its transport across lung epithelial cells and uptake by macrophages

Mahri, Sohaib; Hardy, El�onore; Wilms, Tobias; De Keersmaecker, Herlinde; Braeckmans, Kevin; De Smedt, Stefaan; Bosquillon, Cynthia; Vanbever, Rita

Abstract

Conjugation to high molecular weight (MW ≥ 20 kDa) polyethylene glycol (PEG) was previously shown to largely prolong the lung residence time of recombinant human deoxyribonuclease I (rhDNase) and improve its therapeutic efficacy following pulmonary delivery in mice. In this paper, we investigated the mechanisms promoting the extended lung retention of PEG-rhDNase conjugates using cell culture models and lung biological media. Uptake by alveolar macrophages was also assessed in vivo. Transport experiments showed that PEGylation reduced the uptake and transport of rhDNase across monolayers of Calu-3 cells cultured at an air–liquid interface. PEGylation also decreased the uptake of rhDNase by macrophages in vitro whatever the PEG size as well as in vivo 4 h following intratracheal instillation in mice. However, the reverse was observed in vivo at 24 h due to the higher availability of PEGylated rhDNase in lung airways at 24 h compared with rhDNase, which is cleared faster. The uptake of rhDNase by macrophages was dependent on energy, time, and concentration and occurred at rates indicative of adsorptive endocytosis. The diffusion of PEGylated rhDNase in porcine tracheal mucus and cystic fibrosis sputa was slower compared with that of rhDNase. Nevertheless, no significant binding of PEGylated rhDNase to both media was observed. In conclusion, decreased transport across lung epithelial cells and uptake by macrophages appear to contribute to the longer retention of PEGylated rhDNase in the lungs.

Citation

Mahri, S., Hardy, E., Wilms, T., De Keersmaecker, H., Braeckmans, K., De Smedt, S., …Vanbever, R. (2021). PEGylation of recombinant human deoxyribonuclease I decreases its transport across lung epithelial cells and uptake by macrophages. International Journal of Pharmaceutics, 593, Article 120107. https://doi.org/10.1016/j.ijpharm.2020.120107

• Journal Article Type: Article
• Acceptance Date: Nov 18, 2020
• Online Publication Date: Nov 28, 2020
• Publication Date: Jan 25, 2021
• Deposit Date: Dec 11, 2020
• Publicly Available Date: Nov 29, 2021
• Journal: International Journal of Pharmaceutics
• Print ISSN: 0378-5173
• Electronic ISSN: 1873-3476
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 593
• Article Number: 120107
• DOI: https://doi.org/10.1016/j.ijpharm.2020.120107
• Keywords: Recombinant human deoxyribonuclease I; PEGylation; Pulmonary delivery; Alveolar macrophages; Transport across Calu-3; Diffusion in mucus
• Public URL: https://nottingham-repository.worktribe.com/output/5141126
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0378517320310929?via%3Dihub
• Additional Information: This article is maintained by: Elsevier; Article Title: PEGylation of recombinant human deoxyribonuclease I decreases its transport across lung epithelial cells and uptake by macrophages; Journal Title: International Journal of Pharmaceutics; CrossRef DOI link to publisher maintained version: https://doi.org/10.1016/j.ijpharm.2020.120107; Content Type: article; Copyright: © 2020 Elsevier B.V. All rights reserved.