Search bioRxiv⌕ Search

bioRxiv · 10.1101/2025.06.30.662440

Human pluripotent stem cell-derived bronchial airway organoids provide insights into differential innate immune and long-term responses to SARS-CoV-2 infection in healthy and COPD.

Abstract

Respiratory infections are a major global health concern, as underscored by the COVID-19 pandemic. To better understand bronchial tissue responses to viral infection, we have developed a preclinical in vitro model mimicking the multiciliated airway epithelium, from induced pluripotent stem cell (iPSC) and cultured in an air-liquid interface (iALI). By using iPSCs reprogrammed from patients with chronic obstructive pulmonary disease (COPD), we successfully generated a fully differentiated and functional bronchial epithelium exhibiting key COPD features with goblet and basal cell hyperplasia and tissue inflammation. SARS-CoV-2 could infected and replicated for several weeks in both healthy and COPD models, with a recurrent peak at 3 days after infection. Infected iALI exhibited cilia destruction and increased mucus secretion. Innate immune response of different infected iALI reveals a differential expression of interferon-stimulated genes (ISGs) and pro-inflammatory cytokine secretion. Notably, COPD iALI displayed an earlier innate immune response to SARS-CoV-2 infection as compared to healthy iALI, suggesting a genetic susceptibility of COPD iALI towards inflammation induced by SARS-CoV-2 infection, and a less efficient response to antivirals. In conclusion, our study demonstrates that the iALI bronchial organoid model is a powerful tool for investigating bronchial tissue responses to long term respiratory viral infections, antivirals, and patients with COPD or other airway pathology. Grapical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=151 SRC="FIGDIR/small/662440v1_ufig1.gif" ALT="Figure 1"> View larger version (53K): org.highwire.dtl.DTLVardef@9e5ef7org.highwire.dtl.DTLVardef@16ccf60org.highwire.dtl.DTLVardef@a2b29dorg.highwire.dtl.DTLVardef@17c6f14_HPS_FORMAT_FIGEXP M_FIG C_FIG HighLightsO_LIhiPSC-derived COPD airway organoids C_LIO_LISARS-CoV-2 productively infects induced pluripotent stem-cell derived bronchial organoids iALI and persisted over the long term C_LIO_LISARS-CoV-2 iALI infection results in cilia destruction, increased mucus secretion and a strong innate immune response C_LIO_LISARS-CoV-2 infection elicited a higher and earlier innate immune response in iCOPD C_LIO_LISARS-CoV-2 infection in iCOPD respond less to antivirals C_LI Short AbstractSARS-CoV-2 causes severe lower respiratory tract infection in COVID-19 patients, which can persist over time. Here, we used an in-house developed in vitro airway organoid derived from induced human pluripotent stem cells (iALI) to study SARS-CoV-2 infection over long term in healthy or COPD patients whom respiratory failure is at risk during infection. Our results show that SARS-CoV-2 infection results in high and lethal infection of bronchial epithelial cells, that persist over time, inducing mucus secretion, destruction of ciliated cells and specific cytokine release. A late innate immune response is observed in the healthy iALI, while in iCOPD, it appears earlier and stronger, suggesting a different sensing of SARS-CoV-2 in COPD patients, accompanied by a reduce sensitivity to antivirals. In conclusion, our study demonstrates that the iALI organoid model is a powerful tool for investigating bronchial tissue responses to long term respiratory viral infections, from healthy to pathologic patients.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Morichon, L., SWAIN, J., Gros, N., Nasri, A., Foisset, F., Galisot, G., Racine, V., ASSOU, S., Bourdin, A., de Vos, J., MURIAUX, D.. 2025-07-02. Human pluripotent stem cell-derived bronchial airway organoids provide insights into differential innate immune and long-term responses to SARS-CoV-2 infection in healthy and COPD.. https://doi.org/10.1101/2025.06.30.662440

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

HUH-tag conjugation enables accessible, site-specific functionalization of DNA origami with signaling-competent proteins

DNA origami enables precise nanometer scale control over the number, spacing, and geometry of biomolecules, making it a powerful tool for dissecting spatially organized biological processes such as receptor signaling. However, functionalizing origami with proteins remains a bottleneck restricting the technology to specialized laboratories. Here we show that viral-derived HUH endonuclease tags, small (~10 kDa) domains that form a covalent phosphotyrosine bond with short unmodified ssDNA, provide a simple and cost-effective route to decorating DNA origami with functional proteins. Using interleukin-1{beta} (IL-1{beta}) as a model ligand, we generated recombinant IL-1{beta}-HUH, achieved efficient site-specific conjugation to unmodified ssDNA within minutes, and coupled the conjugates to a DNA origami pegboard with tunable stoichiometry. IL-1{beta}-functionalized origami triggered ligand-dependent signalling in living cells and allowed visualization of the recruitment of downstream signaling molecules, confirming that HUH-tag conjugation preserves functionality. Because this strategy requires only standard molecular biology equipment and unmodified DNA, it lowers a technical and financial barrier to DNA origami-based experimentation, opening the approach to a broader range of biological questions and laboratories outside the DNA nanotechnology field.

cell biology↗

Global mapping of VEGF-A driven changes in the RNA bound phospho-proteome reveals a molecular switch to coordinate angiogenesis in endothelial cells

RNA binding proteins (RBPs) serve as upstream master regulators of cellular fate transitions at the post-transcriptional level. RBP-RNA interactions are dynamically regulated by post-translational modifications to coordinate cellular responses. We hypothesised that changes in RBP phosphorylation will play a key role in coordinating angiogenesis, the formation of new blood vessels, in endothelial cells (ECs). Therefore, we used phospho-proteomic analysis of the mRNA bound proteome to identify global phosphorylation changes in mouse cardiac ECs during acute and prolonged VEGF-A stimulation. Of the differentially phosphorylated RBPs, we found that the phosphorylation of serine 199 within mRNA bound Phf6 (PHD finger protein 6) was consistently increased following VEGF-A stimulation. Phf6 phosphorylation at this specific site was sufficient and required to induce a pro-angiogenic phenotype, facilitating a change in Phf6 conformation promoting interaction with distinct RNAs. We show here that Phf6 phosphorylation at serine 199, downstream of VEGF-A, acts as a molecular switch to differentially coordinate a decrease of anti-angiogenic RNAs (i.e. extracellular matrix genes) and an increase in pro-angiogenesis related RNAs, in part by differentially regulating their stability, to promote angiogenesis.

cell biology↗

Metaplasia Orchestrates Tissue Recovery and Immune Remodeling Through CD73 in Chronic Pancreatitis

Chronic inflammatory injury promotes epithelial metaplasia, and progression from metaplasia to dysplasia is a recognized initiating event in carcinogenesis, as metaplastic cells exhibit heightened susceptibility to genetic alterations. In the pancreas, persistent acinar-to-ductal metaplasia (ADM) can progress to pancreatic intraepithelial neoplasia (PanIN), yet the mechanisms that determine whether ADM resolves or persists with fibrosis and sustained inflammation remain poorly defined. Here, we identify CD73 as a central regulator of ADM fate, tissue repair, and immune microenvironment remodeling during chronic pancreatic injury. Analysis of previously published single-cell RNA sequencing dataset from chronic pancreatitis (CP) tissues identified enrichment of Nt5e, which encodes CD73, within Hnf1{beta} ADM populations. Hnf1bCreERT2 and CD73fl/fl;Hnf1bCreERT2 mice were treated with caerulein for 4 weeks to induce pancreatitis and ADM prior to tamoxifen-induced deletion of CD73 in HNF1B-expressing metaplastic epithelial cells. Histologic phenotyping and immune profiling were performed using immunohistochemistry and imaging mass cytometry. Metaplasia-specific CD73 deletion exacerbated pancreatic injury and fibrosis, impaired epithelial proliferation and recovery of the acinar compartment, and altered the abundance and spatial organization of inflammatory cells. Ex vivo ADM assays further demonstrated that CD73 loss increased ADM formation and the production of MCP-1 and TIMP-1. These findings identify CD73 as a context-dependent checkpoint linking epithelial plasticity and immune modulation during chronic pancreatic injury and tissue repair.

cell biology↗