Shuangshen granules enhance anti-PD1 therapy efficacy in lung adenocarcinoma by modulating myeloid-derived suppressor cell-induced T cell exhaustion

Background Worldwide, lung cancer is the most common cause of cancer-related deaths. Molecular targeted therapies and immunotherapies for non-small-cell lung cancer (NSCLC) have improved outcomes markedly over the past two decades. However, the vast majority of advanced NSCLCs become resistant to current treatments and eventually progress. A traditional Chinese medicine (TCM) formula of Shuangshen granules (SSG) has demonstrated potential in alleviating cancer side effects and imporving survival rate. Despite clinical evidence supporting its benefit, there is still insufficient understanding of the active compounds in SSG and their underlying mechanisms, which limits its broader clinical application. Objective This study aims to identify the key active ingredients in SSG and explore their mechanisms, particularly through modulating myeloid-derived suppressor cell (MDSC)- induced T cell exhaustion, to provide a scientific basis for its application in cancer treatment. Methods The primary active compounds, potential therapeutic targets and intervening signaling pathways, which SSG might inhibit lung adenocarcinoma (LUAD) were predicted by network pharmacology and molecular docking. Subsequently, Lewis lung carcinoma (LLC) tumor-bearing mouse model was established to assess the efficacy of combined SSG and anti-PD-1 therapy in vivo, and MDSCs and CD8⁺T cells were isolated for in vitro co-culture experiments, while pathological examination was conducted using hematoxylin and eosin (HE). The expression of PD-1, TIM-3, CTLA-4, LAG-3 Arg-1, IDO, iNOS, PD-L1 and Gal-9 was detected using immunohistochemistry (IHC), immunofluorescence, and flow cytometry and Western blotting. The expression of IL-2, TNF-α and IFN-γ were detected by reverse transcription-quantitative polymerase chain reaction (qPCR). Concentrations of IL-10 and TGF-β were measured by enzyme-linked immunosorbent assay (ELISA) Results We obtained 19 active ingredients of SSG and predicted 37 potential targets through network pharmacology analysis, among which MOL001792, MOL000449, MOL000358 and MOL000098 were selected as core drug ingredients, and EGFR, IL1B, IL6 and TNF were identified and included into the range of core targets. GO and KEGG analyses suggested that the TNF signaling pathway might hold a crucial role in lung cancer by reducing MDSC and T-cell exhaustion. In the animal experiment, SSG increased TNF-α levels and reduction of T cell exhaustion markers and the down-modulation of MDSC suppressive mediators. Conclusion This investigation identifies MOL001792, MOL000449, MOL000358 and MOL000098 as critical active ingredients in SSG, impacting key biomarkers such as EGFR, IL1B, IL6 and TNF. These substances effectively modulate the TNF signaling pathway, alleviating MDSC-induced T cell exhaustion and restoring anti-tumor immune function.