Research Of The Clinical And Biology Features In Philadelphia-Like Acute Lymphoblastic Leukemia

Three chapters will be concluded in this thesis.Chapter One:Optimization of diagnostic process and related biological characteristics of Ph-like acute lymphoblastic leukemia;Chapter Two:The clinical characteristics and prognostic significance of CRLF2 gene expressions in B-acute Lymphoblastic Leukemia;Chapter Three:Molecular and cytogenetic characterization of four Philadelphia-Like Acute Lymphoblastic Leukemiapatients with EPOR rearrangement.Chapter One:Diagnostic procedures and related studies of Ph-like acute lymphoblastic leukemiaObjective:To establish a rapid and convenient clinical routine diagnostic procedure for Ph-like acute lymphoblastic leukemia,and analysis the clinical and laboratory characteristics of twenty two Ph-like acute lymphoblastic leukemia patients.Methods:B-ALL patients from Jan.2013 to Sep.2018 were first screened by flow cytometry,and patients with BCR-ABL1,ETV6-RUNX1,TCF3-PBX1 and MLL rearrangement excluded by standard cytogenetic R-banding analysis and multiple PCR,then Ph like acute lymphoblastic leukemia cases were screened by fluorescence in situ hybridization(FISH)combined with multiple PCR in patients with specimens.In addition nine gene alterations including JAK2,PAX5 and P53 were detected in some patients.Results:1.549 B-ALL patients were screened by flow cytometry.289 patients with BCR-ABL1,ETV6-RUNX1,TCF3-PBX1 and MLL rearrangement were excluded by karyotype analysis,fluorescence in situ hybridization(FISH)and multiplex PCR.At last,22 patients with Ph-like ALL(all FISH screened)were detected by FISH combined quantitative RT-PCR in 175 patients(155 FISH specimens and 20 RNA specimens),22 patients were detected as Ph-like acute lymphoblastic leukemia(ALL),including 7 patients with CRLF2 rearrangement,4 patients with EPOR rearrangement,2 patients with JAK2 rearrangement,5 patients with PDGFRB rearrangement,3 patients with ABL1 rearrangement,and 1 patient with CSF1R rearrangement.2.Among the 22 patients,there were 16 males and 6 females.the median age was 23.5 years,the median survival was 14.2 months,5 patients relapsed,and 9 patients did not achieve complete remission after the first induction.The largest number of Ph-like ALL patients was in the 21-39 age group(45%).Such patients were negatively correlated with age of onset and the first remission rate(P=0.047,P=0.029),and positively correlated with bone marrow blasts(P=0.0348).3.The proportion of JAK2 mutation,IK6 positive and CRLF2 expression in Ph-like ALL group were significantly higher than those in non-Ph-like ALL group(5/22,22.7%vs 2/153,1.3%,P<0.0001;6/22,27.3%vs 14/153,9.2%,P=0.012;7/22,31.8%vs 18/153,11.8%,P=0.007).4.There were statistically significant differences in the overall survival rate(OS)and remission duration(RD)between the Ph like ALL and non-ph like ALL groups(P=0.002,P<0.0001).Conclusion:1.In this study,a feasible diagnostic procedure was established clinically by using multiple RCR methods combined with FISH.2.Among the 22 patients,CRLF2 rearrangement was the most,accounting for 32%.3.Ph-like ALL is a new high-risk subtype.It is younger in age and more common in men.MRD still persists after chemotherapy,and it is easy to relapse with poor prognosis.Early treatment with targeted drugs TKI and CART combined transplantation is expected to improve the prognosis of patients.ChapterTwo:The clinical characteristics and prognostic significance ofCRLF2 gene expressions in B-acute Lymphoblastic LeukemiaObjective:In order to investigate the clinical features and prognostic significance of CRLF2 genes expression in acute lymphoblastic leukemia patients.Methods:CRLF2 gene expressions were measured in 176 ALL cases from Jan.2013 to Sep.2018.Some patients were further tested for 13 gene alterations including FLT3,JAK2,KRAS and P53.The clinical,cytogenetic and molecular characteristics of ALL with CRLF2 expression were summarized.Results:1.176 patients,93 males and 83 females,with a median age of 27 years,were divided into high expression group(32/176,18.2%)and low expression group(144/176,81.8%).A significant association between high CRLF2 expression and mutations of the JAKISTAT pathway(p<0.0001):33.3%(8/24)CRLF2-high samples carried JAK/STAT pathway mutations,while only 7.6%(9/119)CRLF2-low cases harbored these mutations.2.Compared with low CRLF2 expression,high expression of CRLF2 gene was associated with positively correlated with age(P=0.047);JAK2 mutation(P<0.0001)were observed more frequent in high CRLF2 group.3.Overall survival was decreased in high CRLF2 expression(P=0.021)and TP53 mutation(P=0.008)ALL patients;Univariate analysis showed that JAK2 mutation(P=0.003)and high CRLF2 expression(P=0.006)had significant differences.Multivariate analysis revealed that JAK2 mutation(P=0.026)and high CRLF2 expression(P=0.044)had significant differences.4.According to the CRLF2 expression level,Ph-like ALL patients were divided into the CRLF2 high-expression group and the low-expression group.Statistical analysis showed that the OS of the former was shorter than that of the latter,and there was statistical difference(P=0.01 1).Conclusion:1.A significant association between high CRLF2 expression and mutations of the JAK/STAT pathwayin ALL.2.High expression of CRLF2 gene,JAK2 mutation and TP53 mutation are associated with poor prognosis,which can be used for clinical prognosis evaluation of ALL patientsand may refine their molecular risk classification.Chapter Three:Molecular and cytogenetic characterization of four Philadelphia-Like Acute Lymphoblastic Leukemia patients with EPOR rearrangement.Objective:Analysis the clinical and laboratory characteristics of four ph-like acute lymphoblastic leukemia with EPOR rearrangement.Methods:Bone marrow cells were cultured for 24 h and analyzed for standard cytogenetic R-banding.EPOR and IGH genes rearrangement were detected by FISH.Quantitative PCR was used to detect EPOR expression in 1 patient.The efficacy and survival of treatment were followed up.Results:EPOR rearrangement was detected by FISH in all the 4 patients,and IGH rearrangement was found in 1 patient.Quantitative PCR was performed in 1 patient to confirm the high expression of EPOR.In this group,Among three patients received only chemotherapy,2 of whom died,one died with a survival of 6 months and 27.4 months respectively.And another lost to follow-up after 5.5 months.one patient was treat with ruxolitinib?CART and HSCT,He was still alive after 14.6 months.Conclusion:Ph-like acute lymphoblastic leukemia with EPOR rearrangement is a high-risk subtype,EPOR-IGH fusion can be formed by t(14;19)translocation.Patients with such abnormalities are often refractory,prone to relapse,male predominance,poor prognosis et al,but the combination treatment with ruxolitinib can improve the prognosis of patients.