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  3. PPARγ-responsive luciferase reporter system for high-throughput screening of chemical toxins with potential pulmonary fibrosis effects

PPARγ-responsive luciferase reporter system for high-throughput screening of chemical toxins with potential pulmonary fibrosis effects

  • Ecotoxicol Environ Saf. 2025 Nov 15:307:119433. doi: 10.1016/j.ecoenv.2025.119433.
Xueyang Lin 1,  Yi Yang 2,  Yangyang Sun 2,  Weijie Yang 2,  Shengran Wang 2,  Weidong Li 2,  Zhenghao Bao 2,  Ziqi Cui 2,  Yufeng Yang 1,  Simin Lang 2,  Chen Yang 2,  Weiqiang Sun 2,  Xin Sui 2,  Wenya Feng 2,  Xianli Du 2,  Jun Yang 2,  Yongan Wang 3,  Yuan Luo 4
Affiliations

Affiliations

  • 1 State Key Laboratory of Toxicology and Medical Countermeasures, Beijing Institute of Pharmacology and Toxicology, Beijing 100080, China; School of Pharmacy, Qingdao University, Qingdao, Shandong 266071, China.
  • 2 State Key Laboratory of Toxicology and Medical Countermeasures, Beijing Institute of Pharmacology and Toxicology, Beijing 100080, China.
  • 3 State Key Laboratory of Toxicology and Medical Countermeasures, Beijing Institute of Pharmacology and Toxicology, Beijing 100080, China. Electronic address: [email protected].
  • 4 State Key Laboratory of Toxicology and Medical Countermeasures, Beijing Institute of Pharmacology and Toxicology, Beijing 100080, China. Electronic address: [email protected].
PMID: 41273832 DOI: 10.1016/j.ecoenv.2025.119433
Abstract

Background: Pulmonary fibrosis (PF) is a progressive and fatal lung disorder, and emerging evidence suggests that dysregulation of Peroxisome Proliferator-activated Receptor gamma (PPARγ) plays a critical role in its pathogenesis. However, high-throughput screening (HTS) models for identifying fibrogenic chemicals targeting PPARγ remain underdeveloped. This study aimed to establish a reliable HTS cellular model using a PPARγ-responsive luciferase reporter system to rapidly identify chemicals that induce PF via PPARγ dysregulation.

Methods: Hub genes associated with PF were identified through bioinformatics analysis. A stable HTS cellular model was constructed using a lentiviral vector carrying a luciferase reporter gene under the control of PPARγ-responsive elements. The model was validated using RT-qPCR and luciferase assays. To evaluate its accuracy, four known PF-inducing chemicals and ten non-PF chemicals were tested. Furthermore, to evaluate the model's predictive capability for compounds with uncharacterized PF risk, seven anti-cancer compounds were screened. Molecular docking (MOE) and surface plasmon resonance (SPR) were employed to confirm interactions between the identified chemicals and PPARγ.

Results: PPARγ was identified as the key hub gene linked to PF. The PPARγ-responsive cellular model exhibited significantly elevated PPARγ mRNA levels and luciferase activity compared to controls. PF-inducing chemicals suppressed luciferase activity, whereas non-PF chemicals had no effect. Notably, screening of anti-cancer compounds revealde a subset that markedly suppressed PPARγ activity. Molecular docking and SPR analyses demonstrated concentration-dependent binding affinities between PF-inducing chemicals and PPARγ.

Conclusion: This study developed a high-throughput luciferase-based PPARγ-responsive cellular model to screen chemicals that may contribute to PF through the inhibition of PPARγ activity. The model demonstrates potential as a useful tool for preliminary chemical evaluation. These results suggest a possible role of PPARγ-mediated mechanisms in the development of PF. This work may provide a foundational framework for future efforts in hazard screening and drug discovery aimed at mitigating the effects of PF.

Keywords

Adverse outcome pathway (AOP); High-throughput screening (HTS); Luciferase reporter; PPARγ; Pulmonary fibrosis.

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