Part 1:Application Of Liquid Biopsy To Analyze Therapeutic Response And Potential Biomarkers Of Advanced Esophageal Carcinoma Treated With Anti-PD-1 Antibodies Part 2:Revealing Molecular Changes In Chronic Myeloid Leukemia During Targeted Therapy By Singl

Background and Aims:Esophageal carcinoma is a lethal cancer requiring clinical monitoring and timely improvement of treatment options due to aggressive progression and therapeutie resistance.ctDNA is a promising teehnique for predicting prognosis and therapeutic efficacy.However,ctDNA as a marker for patients with advanced Esophageal carcinoma treated with anti-PD-1 is not well established.In this study,we aim to investigate plasma ctDNA as a biomarker for immunotherapy responses of patients with advanced Esophageal carcinoma.Methods:Tumor mutational burden(TMB)is associated with immunotherapy responses.Howeverwwhether blood TMB(bTMB)is associated with clinical outcomes of imrnunotherapy for Esophageal carcinoma remains unclear.We evaluated bTMB with 1280panel and further validated the feasibility of bTMB as a biomarker for inmunotherapy.We performed HLA class I genotyping and evaluated whether HLA heterozygosity was associated with immunotherapy responses and prognosis.We performed a ctDNA follow-up analysis for patients with posttreatment samples who also had detectable ctDNA variants in their baseline samples.We also compared the consistency of mutations in patient tissues and plasma.Result:The Spearman correlation coefficient between 1280-based bTMB and WES-based TMB reached 0.5037.The remission rate of 61.5%(8/13)in the high bTMB group(bTMB>8),and the remission rate of 47.1%in the low bTMB group(bTMB?8).The remission rate of anti-PD-1 antibody immunotherapy in patients who were heterozygosity at HLA was 85.71%,while the remission rate of patients who were homozygosity at HLA was 27.27%.The median PFS of patients who were heterozygosity at HLA was 7.683 months(95%Cl:1.884-9.188),the median PFS of patients who were homozygosity was 1.867 months(95%Cl:0.1088-0.5423),p-0.0022.We compared the concordance of mutated genes between tissues and plasma samples,and found that 38.53%of TMB was only in the blood,26.15%of TMB was only in the tissue,and 35.32%of TMB were both in tissue and blood.ctDNA levels are associated with immunotherapy responses in most patients.We also compared plasma samples of baseline and post-recurrence,and found that 31.38%of the mutant genes were only in the baseline plasma,36.82%of the new genes appeared after recurrence,and 31.8%of the genes presenced in the whole disease process.Conclusions:bTMB were associated with tTMB.There were more patients benefitted from immunotherapy in the high TMB group compared to patients who in the low TMB group.Compared to patients who were homozygosity at HLA?the immunotherapy response rate and PFS of patients who were heterozygosity at HLA were significantly improved and prolonged,suggesting that HLA typing was a potential biomarker for predicting the efficacy of immunological checkpoint blockade therapy.Tumor heterogeneity may be responsible for the difference mutations in tissue and plasma.ctDNA levels are associated with immunotherapy responses in most patients,suggesting that ctDNA is an effective tool for clinical monitoring.Background and Aims:The treatment of chronic myeloid leukemia(CML),a disease caused by t(9;22)(q34;qll)reciprocal translocation,has been largely advanced through application of targeted tyrosine kinase inhibitors(TKI).However,some of the patients failed to benefit from the targeted therapy and progressed to advanced stages.The underlying mechanism,or the difference of the immune structures between responders and non-responders had not been fully elucidated.Due to advances in the past few years,single cell RNA sequencing techniques have enabled the identification of tumor heterogeneity and subclonal populations.We aim to reveal molecular changes in chronic myeloid leukemia during targeted therapy by single-cell RNA sequencing.Methods:Here we applied single cell RNA-sequencing to a total of 62914 cells from 4 patients with differed prognosis and 1 healthy donor,covering different stages during the disease development,we monitored the immune cell structure shift alongside the CML progress,and characterized the signatures of the immune response to the TKI application.Result:Among all the leukemic clusters,8 clusters intensively expressed high-level erythroid markers,and the absolute number of transcripts was much higher than that in common clusters.3 of the clusters showed obvious sample-specific concentration:cluster 10,19 and 14.These progenitor cell populations exhibited overwhelming enrichment in BT cells.Especially,patients with poor prognosis had distinguishable higher fraction of these early-staged cells,suggesting that presence of these clusters could potentially predict the prognosis of Imatinib treatment.For all patients,Imatinib treatment had an obvious positive effect in the restoration of the normal peripheral blood functional structures.However,the 2 patients with favorable prognosis had significantly higher fractions of CD8 T cells or NK cells while a depressed portion of monocytes in comparison with the healthy donor,suggesting a differentiation bias towards T cells lineage resulted by TKI applieation.Conclusions:The reconstitute cell landscapes demonstrated that the leukemic cells in all patients exhibited classical features of erythroid differentiation,and poor-responders to Imatinib treatment had patient-specific pre-treatmentemerged primitive cells in peripheral blood compared with the responders.Besides,Imatinib treatment significantly altered cell population composition in both responders and non-responders,and affected the functions of the immune cells,which could also play vital roles in dispersing responsiveness of the patients.