Resveratrol ameliorates sepsis-induced immune dysfunction by targeting GADD45A to maintain hematopoietic stem cell...

To investigate the potential pharmacological mechanisms of resveratrol (RES) in ameliorating sepsis-associated immune dysfunction via the GADD45A-mediated hematopoietic regulatory pathway.

This study integrated network pharmacology, ensemble machine learning, and single-cell transcriptomics. Core targets were screened using an exhaustive framework of 113 algorithm combinations coupled with SHAP interpretability analysis. CIBERSORT was employed to quantify sepsis-induced alterations in the immune microenvironment. Single-cell RNA sequencing (scRNA-seq) was used to localize cell-specific distribution of hub targets, while the scTenifoldKnk virtual knockout algorithm assessed their systematic transcriptional impacts on hematopoietic and immune lineages. Finally, molecular docking and 100-ns molecular dynamics (MD) simulations were performed to evaluate the binding affinity and thermodynamic stability of RES with candidate targets.

Four hub genes (JAK1, GADD45A, MAPK14, and ARG1) were identified, with GADD45A exhibiting the highest predictive weight (SHAP: 0.0350) and robust diagnostic performance. Immune infiltration analysis revealed a significant myeloid shift in sepsis. Single-cell resolution indicated that GADD45A was specifically enriched in myeloid subclusters, and its expression was highly correlated with the differentiation trajectory of hematopoietic stem cells (HSCs) toward the myeloid lineage. Virtual knockout simulations predicted that GADD45A impairment might perturb transcriptional patterns in HSC-G-CSF and pre-B cells, potentially exacerbating inflammatory responses by weakening the negative regulation of the alarmin S100A9. Although the docking score of RES with GADD45A was not the highest among core genes, MD simulations confirmed superior thermodynamic stability in explicit solvent, providing a predictive structural basis for its role as a pharmacological target.

This study suggests that resveratrol potentially alleviates sepsis-associated myeloid bias and immune dysfunction by modulating GADD45A-mediated HSC transcriptional programs. These predictive findings offer novel molecular clues and potential mechanisms for the application of resveratrol in maintaining hematopoietic homeostasis.