Supplementary file 1_Mapping DEHP-induced metabolic dysregulation through multi-omics and systems biology: an adverse outcome pathway network approach.docx

Integrating experimental and literature evidence identified key event relationships proposed to link early molecular perturbations to downstream adverse metabolic outcomes, including hepatic steatosis, insulin resistance, non-alcoholic fatty liver disease (NAFLD), and obesity. Central mechanistic features of the network converge on lipid remodeling, mitochondrial dysfunction, oxidative stress, serine/S1P pathway disruption, and impaired insulin signaling.

Because the individual omics contrasts are based on three replicates per condition, statistical support in this work is evaluated at pathway rather than single-feature level: under Benjamini-Hochberg control almost no individual transcript is significant, whereas the pathways carrying the mitochondrial and lipid-handling key events remain significant in competitive gene-set tests. Cross-system comparison showed that suppression of oxidative-energy metabolism is the only module conserved across all three systems, whereas PPAR signalling and fatty-acid handling are liver-specific and non-monotonic.

Together, these data support a coherent, evidence-integrating mechanistic hypothesis linking early molecular events to adverse metabolic outcomes across multiple levels of biological organization, rather than a quantitatively validated causal pathway. To our knowledge, this study provides a novel integrative AOPN framework combining multi-omics data across liver and pancreatic models and reveals previously underappreciated metabolic cross-talk, including involvement of serine and S1P pathways.

The proposed framework is explicitly hypothesis-generating and evidence-integrating: it provides a transparent, FDR-audited basis for future quantitative AOP development and supports mechanistically informed chemical risk assessment by identifying early key events, potential biomarkers, and intervention points relevant to human metabolic health.</p>