What question did this study set out to answer?

The research aims to clarify ZEB1’s role in lung fibrosis and assess its potential as a therapeutic target.

May 20, 2026

C71-26 Zeb1 as a Central Regulator of Pro-fibrotic Signaling and Ecm Remodeling in Pathological Fibroblasts

Key Points

The research aims to clarify ZEB1’s role in lung fibrosis and assess its potential as a therapeutic target.
Fibroblasts from IPF and control groups were cultured on various matrices and gels.
Expression levels of microRNAs and proteins were analyzed using scRNA-seq and ChIP-qPCR.
The effect of ZEB1 inhibitors was tested in vivo for anti-fibrotic efficacy.
ZEB1 and miR-96 levels were significantly increased in IPF fibroblasts, while FoxO3a was decreased (p<0.05).
Inhibition of ZEB1 enhanced fibroblast apoptosis and reduced collagen production, confirming ZEB1's regulatory role (p<0.01).
In vivo ZEB1 inhibition notably decreased lung fibrosis, demonstrating its therapeutic potential.

Abstract

Abstract Rationale Idiopathic Pulmonary Fibrosis (IPF) is a progressive interstitial lung disease characterized by the accumulation of apoptosis-resistant fibroblasts and excessive deposition of a pathologically stiff, type I collagen-rich ECM. IPF fibroblasts evade apoptosis through upregulation of microRNA-96 (miR-96), which suppresses FoxO3a that promotes apoptosis susceptibility. ZEB1, a transcription factor implicated in lung fibrosis, regulates the miR-96/FoxO3a axis as well as collagen cross-linking enzymes lysyl oxidase homologs 1 and 4 (LOXL1 and LOXL4). We hypothesize that ZEB1 acts as a central nexus of pro-fibrotic signaling by coordinating fibroblast apoptosis resistance and ECM cross-linking. The goal of this study is to elucidate the role of ZEB1 in the pathogenesis of lung fibrosis and to evaluate its potential as a novel therapeutic target. Methods Control and IPF fibroblasts (n = 5) were cultured on decellularized lung matrices, 3D collagen matrices, silicone gels (0.5 and 64 kPa), and methacrylated gelatin (GelMA) hydrogels. Expression levels of miR-96, FoxO3a, LOXL2, and caspase 3/7 were assessed. scRNA-seq data from three independent sources were analyzed to examine the pathological role of ZEB1 in regulating fibroblast subtypes including the CTHRC1+ fibroblast population. The anti-fibrotic efficacy of a ZEB1 inhibitor and antimiR-96 was then evaluated in an in vivo model. Results In IPF fibroblasts, ZEB1 and miR-96 were markedly increased, while FoxO3a was suppressed when cultured on decellularized lung matrices, 3D collagen, silicone gels of high stiffness (64 kPa), and GelMA hydrogels. The miR-96 promoter contains ZEB1 consensus binding sites, and ChIP-qPCR analysis confirmed ZEB1’s direct binding to this region. Inhibition of ZEB1 in vitro reduced miR-96 and LOXL homolog expression while restoring FoxO3a, leading to enhanced fibroblast apoptosis and decreased collagen production. Analysis of scRNA-seq datasets revealed increased ZEB1 expression in pro-fibrotic IPF fibroblasts, particularly within the CTHRC1+ subpopulation. Finally, pharmacologic inhibition of ZEB1 in vivo significantly reduced lung fibrosis. Conclusions Our findings demonstrate that 1) ZEB1 functions as a critical nexus of pro-fibrotic signaling, integrating soluble and mechanical cues to promote fibroblast resistance to apoptosis and drive ECM remodeling; and 2) targeting ZEB1 attenuates lung fibrosis by enhancing fibroblast apoptosis and suppressing ECM synthesis, cross-linking and stabilization. This abstract is funded by: /NHLBI HL171164

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