Shuangshen granules enhance anti-PD1 therapy effectiveness in lung adenocarcinoma by modulating myeloid-derived
Zhong-Ning He1, Qi Huang2, Yi Li1
1Department of Oncology, Academy of Chinese Medical Sciences, Guang'anmen Hospital, China , Beijing, 100053, China.
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 improving 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.
Methods:
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 myeloid-derived suppressor cells (MDSC) 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). Network pharmacology and molecular docking were utilized to screen for potential therapeutic targets and intervening signaling pathways of SSG in lung adenocarcinoma (LUAD). The predictions derived from this approach were further verified using Western blotting.
Results:
In vivo experiments using LLC xenograft mice demonstrated that SSG suppressed tumor growth in a dose-dependent manner, with high-dose SSG showing optimal efficacy in inhibiting tumor angiogenesis and cell proliferation. SSG enhances anti-tumor immunity by reducing T cell exhaustion and MDSC-mediated immunosuppression, with SSG + anti-PD-1 combination therapy synergistically optimizing the tumor immune microenvironment. Network pharmacology analysis revealed 5 hub targets (IL2, STAT3, HSP90AA1, LGALS3, and FGF2) associated with immune, LUAD, and active ingredients of SSG, with significant enrichment in the PI3K-Akt pathway. Compared with the control group, the protein expression levels of p-PI3K and p-Akt in the SSG group were significantly down-regulated, indicating that the PI3K-Akt pathway may be inhibited.
Conclusions:
SSG could dose-dependently inhibit LLC tumor growth in mice and exert antitumor effects by alleviating T-cell exhaustion and MDSC-mediated immunosuppression. Notably, IL2, STAT3, HSP90AA1, LGALS3 and FGF2, as potential targets of SSG, were significantly enriched in the PI3K-Akt pathway, which provide a novel perspective for the treatment of LUAD.
Insights
Shuangshen granules (SSG) inhibit lung cancer growth by reducing T cell exhaustion and myeloid-derived suppressor cells. This traditional Chinese medicine formula, combined with anti-PD-1 therapy, optimizes the tumor immune microenvironment for treating lung adenocarcinoma (LUAD).
Area of Science:
- Oncology
- Immunology
- Pharmacology
Background:
- Lung cancer remains a leading cause of cancer mortality worldwide.
- While targeted therapies and immunotherapies have improved outcomes for non-small-cell lung cancer (NSCLC), treatment resistance remains a significant challenge.
- Shuangshen granules (SSG), a traditional Chinese medicine, show promise in improving survival and alleviating side effects, but its mechanisms are not fully understood.
Purpose of the Study:
- To investigate the efficacy of Shuangshen granules (SSG) in combination with anti-PD-1 therapy for lung cancer.
- To elucidate the underlying molecular mechanisms of SSG's anti-tumor effects, particularly in relation to the tumor immune microenvironment and potential signaling pathways.
- To identify potential therapeutic targets of SSG in lung adenocarcinoma (LUAD).
Main Methods:
- Established a Lewis lung carcinoma (LLC) mouse model to evaluate SSG and anti-PD-1 combination therapy.
- Utilized network pharmacology and molecular docking to identify SSG's targets and pathways, verified by Western blotting.
- Assessed immune cell populations (myeloid-derived suppressor cells, CD8+ T cells) and immune-related molecules (PD-1, TIM-3, CTLA-4, etc.) using various molecular and cellular biology techniques (IHC, immunofluorescence, flow cytometry, qPCR, ELISA).
Main Results:
- SSG demonstrated dose-dependent inhibition of LLC tumor growth, reduced angiogenesis, and suppressed proliferation in vivo.
- SSG enhanced anti-tumor immunity by mitigating T cell exhaustion and myeloid-derived suppressor cell-mediated immunosuppression.
- Combination therapy synergistically improved the tumor immune microenvironment; network pharmacology identified IL2, STAT3, HSP90AA1, LGALS3, and FGF2 as key targets enriched in the PI3K-Akt pathway, which was inhibited by SSG.
Conclusions:
- SSG effectively inhibits lung cancer growth and exerts antitumor effects by modulating T cell exhaustion and myeloid-derived suppressor cells.
- The identified hub targets (IL2, STAT3, HSP90AA1, LGALS3, FGF2) and the PI3K-Akt pathway provide novel insights into SSG's mechanism of action for lung adenocarcinoma (LUAD) treatment.