Emerging highly pathogenic H5N1 influenza triggers fibrotic remodeling in human airway organoids

The ongoing outbreak of highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.4.4b has affected at least 989 dairy herds across 17 states in the United States (U.S.) and resulted in 70 confirmed human infections, underscoring the urgent need to understand the pathogenesis and therapeutic intervent...

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Main Authors: Hussin Rothan, Ahmed Mostafa, Mahmoud Bayoumi, Chengjin Ye, Ramya S. Barre, Anna Allué-Guardia, Aitor Nogales, Jordi B. Torrelles, Luis Martinez-Sobrido
Format: Article
Language:English
Published: Taylor & Francis Group 2025-12-01
Series:Emerging Microbes and Infections
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Online Access:https://www.tandfonline.com/doi/10.1080/22221751.2025.2532684
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author Hussin Rothan
Ahmed Mostafa
Mahmoud Bayoumi
Chengjin Ye
Ramya S. Barre
Anna Allué-Guardia
Aitor Nogales
Jordi B. Torrelles
Luis Martinez-Sobrido
author_facet Hussin Rothan
Ahmed Mostafa
Mahmoud Bayoumi
Chengjin Ye
Ramya S. Barre
Anna Allué-Guardia
Aitor Nogales
Jordi B. Torrelles
Luis Martinez-Sobrido
author_sort Hussin Rothan
collection DOAJ
description The ongoing outbreak of highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.4.4b has affected at least 989 dairy herds across 17 states in the United States (U.S.) and resulted in 70 confirmed human infections, underscoring the urgent need to understand the pathogenesis and therapeutic interventions of emerging H5N1 viruses. In this study, we modelled infection with a highly pathogenic recombinant human A/Texas/37/2024 H5N1 (rHPh-TX H5N1) strain using human airway organoids (HAO) to investigate viral replication, innate immune response, infection-induced fibrogenesis, and potential therapeutic interventions. rHPh-TX H5N1 replicated efficiently in HAO, eliciting a robust interferon (IFN) response and pro-inflammatory cytokine production. Prolonged infection led to the accumulation of fibroblast-like cells surrounding infected regions, marked by increased alpha-smooth muscle actin (α-SMA) expression and upregulation of transforming growth factor-beta (TGF-β), indicative of fibroblast activation and extracellular matrix (ECM) remodelling. Compared to organoids infected with the pandemic A/California/04/09 H1N1 (pH1N1) strain, rHPh-TX H5N1 induced significantly higher expression of fibrosis-associated markers, including fibronectin (FN), collagen 1A (COL1A), collagen 3A (COL3A), metalloproteinases 2 and 9 (MMP2, and MMP9). Notably, the inhibition of Rho-associated coiled-coil-forming protein kinases (ROCK) signalling reduced fibrogenesis, with ROCK1 inhibition being more effective than ROCK2 inhibition. These findings highlight the potential of targeting ROCK signalling to mitigate H5N1-induced lung fibrosis, informing therapeutic strategies for severe influenza infections.
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spelling doaj-art-6f55a3cf34bc409c850490a48e36c9242025-08-20T03:56:47ZengTaylor & Francis GroupEmerging Microbes and Infections2222-17512025-12-0114110.1080/22221751.2025.2532684Emerging highly pathogenic H5N1 influenza triggers fibrotic remodeling in human airway organoidsHussin Rothan0Ahmed Mostafa1Mahmoud Bayoumi2Chengjin Ye3Ramya S. Barre4Anna Allué-Guardia5Aitor Nogales6Jordi B. Torrelles7Luis Martinez-Sobrido8Host-pathogen interactions (HPI) and Disease Intervention and Prevention (DIP) Programs, Texas Biomedical Research Institute, San Antonio, TX, USAHost-pathogen interactions (HPI) and Disease Intervention and Prevention (DIP) Programs, Texas Biomedical Research Institute, San Antonio, TX, USAHost-pathogen interactions (HPI) and Disease Intervention and Prevention (DIP) Programs, Texas Biomedical Research Institute, San Antonio, TX, USAHost-pathogen interactions (HPI) and Disease Intervention and Prevention (DIP) Programs, Texas Biomedical Research Institute, San Antonio, TX, USAHost-pathogen interactions (HPI) and Disease Intervention and Prevention (DIP) Programs, Texas Biomedical Research Institute, San Antonio, TX, USAInternational Center for the Advancement of Research and Education (I•CARE), Texas Biomedical Research Institute, San Antonio, TX, USACenter for Animal Health Research, CISA-INIA-CSIC, Madrid, SpainInternational Center for the Advancement of Research and Education (I•CARE), Texas Biomedical Research Institute, San Antonio, TX, USAHost-pathogen interactions (HPI) and Disease Intervention and Prevention (DIP) Programs, Texas Biomedical Research Institute, San Antonio, TX, USAThe ongoing outbreak of highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.4.4b has affected at least 989 dairy herds across 17 states in the United States (U.S.) and resulted in 70 confirmed human infections, underscoring the urgent need to understand the pathogenesis and therapeutic interventions of emerging H5N1 viruses. In this study, we modelled infection with a highly pathogenic recombinant human A/Texas/37/2024 H5N1 (rHPh-TX H5N1) strain using human airway organoids (HAO) to investigate viral replication, innate immune response, infection-induced fibrogenesis, and potential therapeutic interventions. rHPh-TX H5N1 replicated efficiently in HAO, eliciting a robust interferon (IFN) response and pro-inflammatory cytokine production. Prolonged infection led to the accumulation of fibroblast-like cells surrounding infected regions, marked by increased alpha-smooth muscle actin (α-SMA) expression and upregulation of transforming growth factor-beta (TGF-β), indicative of fibroblast activation and extracellular matrix (ECM) remodelling. Compared to organoids infected with the pandemic A/California/04/09 H1N1 (pH1N1) strain, rHPh-TX H5N1 induced significantly higher expression of fibrosis-associated markers, including fibronectin (FN), collagen 1A (COL1A), collagen 3A (COL3A), metalloproteinases 2 and 9 (MMP2, and MMP9). Notably, the inhibition of Rho-associated coiled-coil-forming protein kinases (ROCK) signalling reduced fibrogenesis, with ROCK1 inhibition being more effective than ROCK2 inhibition. These findings highlight the potential of targeting ROCK signalling to mitigate H5N1-induced lung fibrosis, informing therapeutic strategies for severe influenza infections.https://www.tandfonline.com/doi/10.1080/22221751.2025.2532684Human airway organoidsinfluenza virusprolonged infectionfibrogenesisROCK signalling pathway
spellingShingle Hussin Rothan
Ahmed Mostafa
Mahmoud Bayoumi
Chengjin Ye
Ramya S. Barre
Anna Allué-Guardia
Aitor Nogales
Jordi B. Torrelles
Luis Martinez-Sobrido
Emerging highly pathogenic H5N1 influenza triggers fibrotic remodeling in human airway organoids
Emerging Microbes and Infections
Human airway organoids
influenza virus
prolonged infection
fibrogenesis
ROCK signalling pathway
title Emerging highly pathogenic H5N1 influenza triggers fibrotic remodeling in human airway organoids
title_full Emerging highly pathogenic H5N1 influenza triggers fibrotic remodeling in human airway organoids
title_fullStr Emerging highly pathogenic H5N1 influenza triggers fibrotic remodeling in human airway organoids
title_full_unstemmed Emerging highly pathogenic H5N1 influenza triggers fibrotic remodeling in human airway organoids
title_short Emerging highly pathogenic H5N1 influenza triggers fibrotic remodeling in human airway organoids
title_sort emerging highly pathogenic h5n1 influenza triggers fibrotic remodeling in human airway organoids
topic Human airway organoids
influenza virus
prolonged infection
fibrogenesis
ROCK signalling pathway
url https://www.tandfonline.com/doi/10.1080/22221751.2025.2532684
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