Systems Pharmacology Dissection Of The Action Mechanism Of Epimedium On Lung Tumor Microenvironment

Background: The clinical notably success development of immunotherapy,potentiating antitumor immunity by activating antitumoral responses of cytotoxic T cells are a promising area of cancer therapy.Natural products that promote antitumor effect via inhibiting inflammation and activating inflammatory immune response,including regulation of the tumor microenvironment(TME).Here,we developed a novel systems pharmacology strategy to analyze the mechanism of polypharmacology molecules of Epimedium,which improves the TME and enhances T cell response in the treatment of non-small cell lung cancer(NSCLC)and conducted antitumor experimental verification.Experimental approach: After screening the polypharmacology molecules of Epimedium,through target prediction and network analysis,we selected the icaritin(ICT),a major active ingredient of Epimedium,with relatively high degree and good pharmacological properties,to carry in vivo and in vitro antitumor experiments.In the experimental verification,the pharmacological effects of Epimedium on NSCLC were analyzed by tumor modeling,transcriptome sequencing,immunofluorescence,western blotting,real-time quantitative PCR and flow cytometry.Research results: In vivo,we evidenced that the ICT effectively inhibited tumor growth in LLC tumor-bearing mice and prolonged mouse survival.In tissue,we performed transcriptome sequencing of ICT-treated tumor,which revealed significantly enrichment of immunobiological process and up-regulation of T-cell chemotactic factors(CXCL9 and CXCL10).In addition,immunofluorescence showed that increased infiltration of CD8+ T cells in ICT-treated tumor.Finally,at the cellular level,we demonstrated that ICT has anti-inflammatory and pro-apoptotic effect on lung cancer cell lines by western blotting,RT-PCR and flow cytometry.Implications: Overall,the approach developed in this study offered an important paradigm for understanding the mechanism of polypharmacology of natural products targeting TME to treat cancer,which will promote the development and application of multi-target natural products in clinical practice.