Plasma Bioactive Factors Profiles Correlated With Clinical Benefit Of Immune Checkpoint Inhibitor In Advanced Non-Small Cell Lung Cancer

BackgroundCancer is one of the major public health problems currently endangering human health.Lung cancer has a very high incidence and mortality rate worldwide and is one of the leading causes of death among all malignancies.According to pathological classification,the most common type of lung cancer is non-small cell lung cancer(NSCLC),which accounts for 85%of all lung cancers.Many patients are already in stage III-IV at the time of diagnosis and have a very poor prognosis,with a 5-year survival of only 16%.Platinum-based chemotherapy regimens have poor remission rates and insignificant survival extension.Although there is some benefit from molecularly targeted therapy for driver mutation positivity,patients with driver mutations are a minority and most patients are prone to relapse within one year.With deeply research on the mechanism of NSCLC development and the understanding of autoimmune surveillance to identify and kill lung cancer cells,the treatment of advanced NSCLC has evolved from chemotherapy to molecular targeted therapy and then to the current hot tumor immunotherapy,forming a pattern of multiple therapeutic approaches.Among them,Immune checkpoint inhibitors(ICIs),as an emerging anti-tumor therapy,have achieved remarkable efficacy in NSCLC and several tumors since their introduction.Tumor cells can release tumor cell-associated antigens during the death process,which are processed by dendritic cells and antigen-presenting cells to activate T cells,resulting in cytotoxic T cells that can infiltrate into the tumor site and recognize and kill tumor cells.In this process,the programmed death receptor-1(PD-1)and its ligand(PD-L1)signaling pathway play an important role.PD-L1 expressed on tumor cells can bind to PD-1 expressed on T cells,natural killer cells and other immune cells to induce immune cell deactivation or apoptosis.Currently,PD-1/PD-L1 inhibitors have been widely used in patients with advanced NSCLC,and tumor tissue PD-L1 expression level has been used as a predictor of anti-PD-1/PD-L1 efficacy,but in practical clinical use,it has been found that PD-L1 expression is not associated with clinical benefit in some cancer types,and even some patients with negative PD-L1 expression are able to benefit from PD-1/PD-L1 inhibitor therapy.Currently,there are many challenges in PD-L1 detection from sampling to interpretation of expression threshold,therefore,more and more studies are devoted to find the markers that can effectively predict the efficacy of ICIs.Objectives1.To analyze the relationship between plasma bioactive factors and the efficacy of immunotherapy in patients with unresectable stage III-IV non-small cell lung cancer by testing plasma samples.And to explore the potential pathological characteristics of the population benefited from immunotherapy through the analysis of typical clinical cases.2.To explore the factors potentially affecting the secretion of soluble cytoactive substances by tumor-associated fibroblasts through in vitro simulation experiments.Methods1.The plasma levels of cytokines/chemokines,depurinic/depyrimidinic endonuclease(APEX1),and transforming growth factor-(Transforming Growth Factor-β,TGF-β)were measured by ELISA kit and Bio-Plex Pro Human Cytokines Grp I Panel(27-plex)inunresectable stage III-IV non-small cell lung cancer patients before and 12 weeks after treatment with PD-1/PD-L1 inhibitors.APEX1,TGF-β expression levels in plasma before and12 weeks after treatment to compare sustained clinical benefit with no sustained clinical benefit,PD-1/PD-L1 The expression of cytokines/chemokines,APEX1 and TGF-β in the plasma of patients in each group was compared between continuous clinical benefit and no continuous clinical benefit,PD-1/PD-L1 single agent and PD-1/PD-L1 combination chemotherapy.2.To detect the composition of immune cells in postoperative tumor tissue specimens of patients receiving typical clinical benefit cases by multiplex immunohistochemistry(m IHC)and to analyze the characteristics of tumor-infiltrating immune cells in patients benefiting from immunotherapy.Results1.In patients with unresectable stage III-IV non-small cell lung cancer treated with PD-1/PD-L1 inhibitors,high baseline IL-6 levels in the ICI monotherapy cohort or elevated levels of IL-6,IL-8,FGF2,CXCL10,CCR1,PDFGB,TNF-α,and APEX1 after treatment were associated with shorter progression free survival(PFS).Elevated CXCL10 levels after treatment were also associated with poorer overall survival(OS).In the ICIs combination therapy group,high levels of baseline IL-12,IL-17 A,FGF2,VEGF and APEX1 and elevated post-treatment CCL2 were associated with poorer PFS.High levels of IL-9,FGF2,PDFGB,CCL4,TFGB,and APEX1 before treatment and elevated IL-13,CSF2,and CCL2 after treatment were associated with poor OS in combination therapy patients.A linear regression model was developed to model PFS and OS risk based on multiple cytokines,which could better predict the risk of progression and survival of patients.2.Knockdown of APEX1 in BJ-5ta cells could directly affect the secretory function of IFN-B1,IL-6,IL-1B,CCL5,CXCL10,IL-8,and IL-1R1.Conclusions1.Plasma cytokines/chemokines are relatively non-invasive and feasible biomarkers.In this study,we successfully determined the express of plasma soluble bioactive factors in patients with unresectable stage III-IV non-small cell lung cancer before and after treatment with PD-1/PD-L1 inhibitors,and calculated the rate of change of plasma soluble bioactive molecules after treatment.The correlation with clinical benefit was analyzed and provided a reference cytokines/chemokines profile to predict the prognosis of patients treated with PD-1 /PD-L1 inhibitor monotherapy and patients treated with PD-1/PD-L1 inhibitors combined with chemotherapy,which can be dynamically monitored dynamically at different times of treatment by selecting different combinations according to the treatment regimen of different ICIs.2.In this study,risk models of PFS and OS were constructed based on the baseline levels and change rate of cytokines/chemokines after treatment,by which the prognosis of patients could be better predicted.3.During immunotherapy for squamous lung cancer,changes in the tumor microenvironment can be directly involved in the process of immune-related tumor cell death,which may be caused by the infiltration of immune cells mediated by relevant cytokines and chemokines secreted by fibrous connective tissue.APEX1 can influence the secretion of relevant cytokines and chemokines by fibroblasts in the tumor mesenchyme by affecting APEX1 may affect the efficacy of tumor immunotherapy by affecting the secretion of related cytokines and chemokines in the tumor mesenchyme,providing a potential operational target for clinical improvement of the efficacy of PD-1 / PD-L1.