Mutational Spectrum And Prognostic Stratification Of AML Based On Next Generation Sequencing

Acute myeloid leukemia(AML)is a kind of genetic heterogeneity disease caused by complex interactions among different carcinogenic factors.Genomics research led to the discovery of molecular pathogenesis of AML and developed a great diversity among various subtypes of AML defined by traditional cytogenetics.Cytogenetics alone is not enough to guide the clinical outcome of AML.In recent years,although NCCN guidelines have been incorporated into several common gene mutations(NPM1、CEBPA、FLT3-ITD、TP53、KIT),it’s still not enough to stratify AML,especially for intermediate-risk AML that accounting for more than 60%,other new genes and combinations of different mutations are needed for further stratification.In addition,genes often have more than one mutation sites,a variety of mutations also occur frequently,single mutation and site has been insufficient to guide clinical outcomes.Based on this,the traditional generation sequencing has been unable to meet demand,new techniques are needed to further study the molecular mechanism of AML and assisted multiple mutational analysis for clinical.Here we applied next generation sequencing(NGS)platform to screen mutational hotspots in 111 genes relevant to hematological malignancy,and analyzed the guiding signification of mutations on phonotype and prognostic stratification by combing clinical features.Firstly,by sequencing data analysis,filtering and validation,we get the unique AML mutation frequency and spectrum of Chinese people.Then,the molecular mechanism of AML and the relationship between mutation and clinical features were studied from two aspects.On the one hand,through single mutational analysis,we identified the relationship between different mutations,mutations and clinical phenotypes(karyotype,age,newly diagnosed white blood cell count,immunophenotype)、clinical efficacy and prognosis(response rate and survival),and confirmed several collaborative and exclusive pattern among different biological function mutations.On the other hand,through the integration of multiple mutation analysis,co-occurrence of FLT3-ITD,NPM1 and DNMT3 A mutations was found commonly and was identified with associated with specific clinical AML features like older age、high WBC count、mononuclear cell morphology and poor outcomes.While co-occurrence of NPM1 and IDH1/2 without FLT3-ITD was iedntified with lower WBC count and was sensitive to chemotherapy.Furthermore,according to 3 different combinations of mutations: that is,NPM1 or bi-CEBPA mutations without others,FLT3-ITD、DNMT3A、ASXL1、TET2、TP53 or PFH6 mutations without CEBPAmut,and other genotype,intermediate-risk patients could be stratified into three distinct risk groups-favorable risk group、intermediate risk group and high risk group,allowing reductions in real intermediate-risk AML by one-third.Our study offered deep insights into the molecular pathogenesis and biology of AML and indicated that the prognosis of IR-AML could be further stratified by different mutation combinations,which layed a foundation for the future development of a new risk stratification system and the realization of AML individual precision treatment.