Investigating the impact of PBAE-mediated FOXJ1 mRNA delivery on differentiation of primary human bronchial epithelial cells
File(s)
Published version
Author(s)
Konstantinidi, Rafaela
Yates, laura
Lloyd, Clare
Saglani, Sejal
Patel, Asha
Type
Journal Article
Abstract
Primary human basal bronchial epithelial cells (HBECs) are an important population of progenitor cells capable of self-renewal and differentiation to maintain airway homeostasis. At air-liquid interface (ALI) culture, HBECs undergo mucociliary differentiation, providing a robust
physiologic model to evaluate novel therapeutics such as in vitro transcribed messenger RNA (IVT-mRNA). However, the impact of IVT-mRNA delivery on the differentiation potential of basal HBECs remains poorly characterised. Poly (beta amino) ester (PBAE) nanoparticles
have demonstrated effective airway delivery of IVT-mRNA in various pre-clinical studies. Here, we aimed to understand the impact of PBAE-mediated mRNA transfection on basal HBEC differentiation at ALI. We investigated IVT-mRNA encoding the ciliogenesis transcription factor Forkhead box J1 (FOXJ1) as a model tool for transient overexpression in primary basal
HBECs and characterised its subsequent impact on epithelial integrity and differentiation at ALI. PBAE-mediated delivery of FOXJ1 mRNA to submerged primary HBECs resulted in approximately 50% FOXJ1-positive cells and transient upregulation of key ciliogenesis-related
genes, including DNALI1 and RSPH9. Following 28 days of differentiation at ALI, FOXJ1 or reporter mRNA transfected cultures displayed normal epithelial morphology, with tight junction and differentiation markers, proportions of secretory and ciliated cells, and cilia ultrastructure comparable to non-transfected controls. These data indicate that PBAE-mediated IVT-mRNA delivery can transiently increase encoded protein expression in basal primary HBECs, without impeding mucociliary differentiation.
physiologic model to evaluate novel therapeutics such as in vitro transcribed messenger RNA (IVT-mRNA). However, the impact of IVT-mRNA delivery on the differentiation potential of basal HBECs remains poorly characterised. Poly (beta amino) ester (PBAE) nanoparticles
have demonstrated effective airway delivery of IVT-mRNA in various pre-clinical studies. Here, we aimed to understand the impact of PBAE-mediated mRNA transfection on basal HBEC differentiation at ALI. We investigated IVT-mRNA encoding the ciliogenesis transcription factor Forkhead box J1 (FOXJ1) as a model tool for transient overexpression in primary basal
HBECs and characterised its subsequent impact on epithelial integrity and differentiation at ALI. PBAE-mediated delivery of FOXJ1 mRNA to submerged primary HBECs resulted in approximately 50% FOXJ1-positive cells and transient upregulation of key ciliogenesis-related
genes, including DNALI1 and RSPH9. Following 28 days of differentiation at ALI, FOXJ1 or reporter mRNA transfected cultures displayed normal epithelial morphology, with tight junction and differentiation markers, proportions of secretory and ciliated cells, and cilia ultrastructure comparable to non-transfected controls. These data indicate that PBAE-mediated IVT-mRNA delivery can transiently increase encoded protein expression in basal primary HBECs, without impeding mucociliary differentiation.
Date Issued
2026-06-01
Date Acceptance
2026-05-17
Citation
Biology Open, 2026, 15 (6)
ISSN
2046-6390
Publisher
The Company of Biologists
Journal / Book Title
Biology Open
Volume
15
Issue
6
Copyright Statement
© 2026. Published by The Company of Biologists This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
License URL
Identifier
10.1242/bio.062419
Subjects
mRNA
Transcription factor
FOXJ1
Primary human bronchial epithelial cells
Air-liquid interface culture
Differentiation Medicover Genetics
Egkomi 2409
Cyprus
Publication Status
Published
Article Number
ARTN bio062419
Date Publish Online
2026-06-12