Exploration Of New Biomarkers Associated With Treatment And Prognosis Of Lung Cancer

Background:Detection of T cells that target tumor-specific or associated mutant antigens is mostly confined to tumor resident lymphocytes.To date,evidence of such T cells in peripheral blood in lung cancer patients is rare.Methods:we assessed the proportion of peripheral CD137⁺or PD-1⁺CD8⁺T cell subsets and CD4⁺CD25⁺CD127^low/-CTLA-4⁺regulatory T cell subset in 83 patients with lung cancer prior to or after treatment and in 49 age-and gender-matched healthy donors as controls,correlated which to clinical characteristics in the lung cancer patient group.we also analysed the CD8 T cells co-express of CD137⁺and PD-1⁺as biomarker exploring,and investigate the correlations between tumor reactive CD8⁺T cell subsets and CTLA-4⁺CD4⁺CD25⁺CD127^low/-regulatory T cells.Results:We observed an increase in peripheral PD-1⁺and CD137⁺CD8⁺T cells in lung cancer patients,which elevated with age,despite treatment or stage.Lung cancer patients contain higher proportions of CD8⁺T cells that co-express both PD-1 and CD137.The proportion of CTLA-4⁺CD4⁺CD25⁺CD127^low/-regulatory T cells positively correlated with that of tumor reactive CD8⁺T cell subsets.In the result of T cell receptor sequencing of two lung cancer patients,the clonal diversity of TCR of CD8⁺PD-1⁺T cells in one patient（small-cell lung cancer,T1N1M0）was lower than the corresponding CD8⁺PD-1^-T cells and the concentration of the frequency of TCR clonoltypes of CD8⁺PD-1⁺T cells were more obvious than CD8⁺PD-1⁺T cells.nevertheless,the same phenomenon was not observed in another patient（lung adenocarcinoma,T1N0M0）.Conclusion:Our result provide evidence for the presence of tumor reactive T cells in the peripheral blood of lung cancer patients,which steadily increased with age,with a strong,positive correlation to the proportion of effector regulatory T cells.These findings provided more information to the timing and patient-selection for immunotherapeutic strategies,Our result supports the immunotherapeutic intervention strategy using combination therapy for differential control of regulatory T cells and activation of tumor reactive T cells.Background: CD137 is activation-dependent and has been reported preferentially expressed on tumor-reactive subset of TILs.Combination modality of immunotherapy with CD137 agonists have shown excellent safety and promising efficacy in early clinical trials.To date,the knowledge of CD137+ tumor infiltrating lymphocytes in tumor tissue of lung cancer patients and the correlations with clinical pathological characteristics and survival of lung cancer are still largely unclear.Methods: In this study,118 lung adenocarcinoma patients of IIIb-IV stage were enrolled,the corresponding tumor tissue samples were collected and representative tumor areas of 2 mm diameter were chosen,tissue microarrays were then constructed and followed by performance of multiplex immunofluorescence stain with CD8,CD137,PD-1,IFN-γand granzyme B.Samples were divided into different intratumoural or stromal groups,based on whether they had higher or lower densities of specific tumor-infiltrating lymphocyte populations in the intratumoral space or stroma.And their correlations with clinical characteristics and survival were analyzed.Results : In our study,most CD137+ cells were CD8 negative.Most CD137+ lymphocytes expressed IFN-γ and granzyme B at a much higher level than CD8+PD-1+ T cells,while the expression of IFN-γand granzyme B levels on CD8+PD-1+ T cells were as low as paired CD8+PD-1-T cells.In the univariate analysis,patients with higher density of CD8+ TILs in the stroma were observed to have better survival than their counterparts both in all the 118 samples and in the EGFR mutant subgroup.No correlation were observed between the proportion of CD137,CD8 or PD-1 positive lymphocytes and gender,age,EGFR mutation status.smokers were observed to habor higher proportion of stroma CD8+ lymphocytes than non-smoker（p=0.023）and have a trend to habor higher proportion of CD137+ lymphocytes than non-smoker（p=0.053）.Conclusions:the observation of CD137 expression in tumor microenvironment could offer further understanding of the feature of CD137+ lymphocytes in advanced lung adenocarcinoma.Our findings may provide more information to the patient-selection of immunotherapies.Background:Subclonal architecture and genomic evolution of small cell lung cancer（SCLC）under treatment has not been well studied primarily due to lack of tumor specimens,particularly longitudinal samples acquired during treatment.SCLC is characterized by early hematogenous spread,which makes circulating cell-free tumor DNA（ct DNA）sequencing a promising modality for genomic profiling.Methods: We performed targeted deep sequencing of 430 cancer genes on pre-treatment tumor biopsies as well as plasma samples collected prior to and during treatment from 22 SCLC patients.We also explored the subclonal architecture of SCLC from ct DNA sequencing,investigated the molecular evolution of SCLC during treatment and assessed the association between ct DNA and clinical parameters and survival.Results: Similar subclonal architecture was observed between pre-treatment ct DNA and paired tumor DNA.Mean variant allele frequency of clonal mutations from pre-treatment ct DNA was associated with progression-free survival and overall survival.Pre-and post-treatment ct DNA mutational analysis demonstrated that mutations of DNA repair and NOTCH signaling pathways are enriched in post-treatment samples.Conclusions: These data suggest that ct DNA sequencing is promising to delineate genomic landscape,subclonal architecture and genomic evolution of SCLC,and might also be used as a potential prognostic biomarker candidate.