| Background:Acquired bone marrow failure syndromes(aBMFS)are a group of heterogeneous disorders that are caused by quantitative and qualitative defects of hematopoietic stem/progenitor cells(HSPCs),and bone marrow failure and underlying cytopenia are the main manifestation.Hematological disorders,such as aplastic anemia(AA),paroxysmal nocturnal hemoglobinuria(PNH)and myelodysplastic syndrome(MDS)are common paradigms of aBMFS in clinical work.Bone marrow failure is the shared feature of these diseases,but each has its specialty in clinical feature and treatment.Besides,each of these diseases can undergo clonal evolution and bear the varying degree risk of transforming into another malignant diseases.Therefore,it is of importance and challenge to diagnose these diseases.WT1,which is located at chromosome 11p13,plays a vital role in the tumorigenesis of Wilms' tumor.Additionally,it has been reported that WT1 plays a substantial role in controlling the growth and differentiation of various immature cell types,including CD34+HSPCs.Recent years,accumulating reports have demonstrated that WT1 can commonly serve as an target indicator for minimal residual disease(MRD)in malignant hematology diseases,however,WT1 expression level has not yet been thoroughly validated on aBMFS.Objective:The study was designed to monitor the dynamic expression level of WT1 in patients with AA,PNH and MDS,explore the effect of WT1 in patients with aBMFS,and take this one step further,to afford novel strategies for differential diagnoses and prognostic evaluation for aBMFS.Methods:(1)A cohort of 387 patients in total were included in our study,there were 180 naive cases with cytopenia which were finally diagnosed with AA(76 cases),PNH(20 cases),and MDS(84 cases).Additionally,there were 189 confirmed cases of AA who had volunteered to receive immunosuppressive therapy(IST)for at least 2 years;the remaining 18 cases were MDS/AML patients who had evolved from AA.(2)housekeeping gene c-ABL worked as reference gene for normalization.Quantitative measurement of WT1 transcript level in bone marrow mononuclear cells was performed using quantitative real-time polymerase chain reaction(qRT-PCR).(3)For intergroup comparison of WT1 mRNA expression,a Mann-Whitney U test and a Wilcoxon rank-sum test were performed.To evaluate the diagnostic accuracy of bone marrow WT1 mRNA,receiver operating characteristic(ROC)analysis was used.Results:(1)The WT1 level in naive cases with aplastic anemia(AA)was significantly lower than that in patients with paroxysmal nocturnal hemoglobinuria(PNH,p=0.023)and myelodysplastic syndrome(MDS,p<0.001).(2)The WT1 level in patients with MDS significantly increased as the disease progressed to an advanced stage.(3)Patients with hypoplastic MDS had a differentiated expression level of WT1 compared with that of non-severe AA(NSAA)(p<0.001).(4)post-treatment patients of AA with partial response(PR)or complete response(CR)status had relatively higher WT1 levels than those with naive AA(p=0.017,p=0.003,respectively).(5)Patients with AA who undergone disease evolution had relatively higher level of WT1 after receiving IST.Conclusion:The expression level of WT1 mRNA in BM varied widely in naive patients with aBMFS.WT1 expression had an overt ascendant trend as disease progressed to each advanced stage.In patients with AA,WT1 expression level fluctuate with disease status.Thus,we can come to conclude that WT1 can serve as a novel parameter for differential diagnoses,curative effect and prognostic evaluation for aBMFS.Background:Acquired aplastic anemia(AA)is a paradigm of bone marrow failure disease,and,immune,especially the autologous T cells mediated quantitative and qualitative defects of hematopoietic stem/progenitor cells(HSPCs)play a vital role in the pathophysiology of AA.Autophagy is a highly conserved process of autodigestion in conditions such as cellular stress,hypoxia,or energy deprivation.Autophagy is closely related to T cell pathophysiology and the destiny of HSPCs,in which autophagy-related gene 5(ATG5)is indispensably involved.The unfortunate fact is that the putative role of ATG5 in AA has not yet been validated.Objective:To determine gene frequencies of ATG5 single nucleotide polymorphisms(SNPs)in AA,we aim to evaluate the putative role of six ATG5 variables(rs3761796,rs473543,rs510432,rs28656919,rs573775,and rs803360)on the susceptibility,severity,and outcome of AA in Chinese population by a case-control study.Methods:The subjects comprised 176 patients with AA,all recruited into the study continuously by the timing of admission,157 age-and sex-matched healthy controls from employees of our hospital.Venous blood samples were collected into 5mL EDTA tubes,genomic DNA was extracted from whole blood.Matrix-assisted desorption/ionization time of flight mass spectrometry(MALDI-TOF/MS)platform to assay the result.All the statistical analyses were performed using Statistical Product and Service Solutions(SPSS).The differences of genotype frequencies of the six ATG5 variables within subgroup of AA,patients and health controls were evaluated,to evaluate the putative role of six ATG5 variables(rs3761796,rs473543,rs510432,rs28656919,rs573775,and rs803360)on the susceptibility,severity,and outcome of AA in Chinese population by a case-control study.Besides,the relationship between occurrence of any new hematological event in patients with AA and six ATG5 gene variants as determined.Results:(1)All genotyped distributions of ATG5 rs3761796,rs473543,rs510432,rs28656919,rs573775,and rs803360 were all in agreement with the Hardy-Weinberg equilibrium for healthy controls.(2)A markedly decreased risk of AA in the recessive models of rs510432 and rs803360 polymorphisms(adjusted odds ratio[OR],95%confidence interval[CI]=0.467[0.236-0.924],p=0.029 for ATG5 rs510432;adjusted OR[95%CI]=0.499[0.255-0.975],p=0.042 for ATG5 rs803360)was observed.(3)The decreased risk was even more pronounced among non-severe AA compared with healthy controls under recessive models(adjusted OR[95%CI]=0.356[0.141-0.901],p=0.029 for ATG5 rs510432;adjusted OR[95%CI]=0.348[0.138-0.878],p=0.025 for ATG5 rs803360;adjusted OR[95%CI]=0.352[0.139-0.891],p=-0.027 for ATG5 rs473543).(4)Above all,rs573775 can strongly predict the occurrence of newly onset hematological event in patients with AA.GG/AG/AA carriers of rs573775 differed dramatically in terms of the occurrence of new hematological events(p=0.007).Compared with non-AA carriers(GG/AG),a higher cumulative rate of new hematological events was observed in patients carried rs573775 AA genotype(p=0.018).Conclusion:MALDI-TOF/MS platform was used to assay the SNPs of ATG5 in patients with AA and health controls,we concluded that in Han-Chinese populations,in the recessive models,rs510432 and rs803360 markedly decreased risk of AA;Besides,decreased risk was even more pronounced among NSAA and controls.Above all,rs573775 can strongly predict the occurrence of newly onset hematological event in patients with AA,AA genotype carriers are more prone to encounter the new hematological events,and patient-tailored medical decision should be made appropriately. |
PDF Full Text Request