Work

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

Year: 2025

Type: article

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.

Source: Ecotoxicology and Environmental Safety

DOI: 10.1016/j.ecoenv.2025.119433