AML-280 a prospective study to evaluate the prognostic implications of methylation and expression of wilms tumor-1 (wt-1) gene in acute myeloid leukemia

Abstract

Context Acute myeloid leukemia (AML) is a genetically heterogeneous hematological disorder distinguished by abnormal differentiation and clonal proliferation of myeloid progenitor cells in the bone marrow. The Wilms tumor-1 (WT-1) gene is a critical regulator of malignant hematopoiesis, which can either activate or repress genes to regulate cellular growth, differentiation, and proliferation of AML. However, its genetic and epigenetic roles remain unknown. Objective This study aimed to investigate the expression levels and methylation status of the WT-1 gene in 143 newly diagnosed cases of AML. Design and Setting This prospective clinical research was performed in our institution from 2020-2023. Patients or Other Participants Bone marrow (BM) and peripheral blood (PB) samples were collected at the time of diagnosis (day-0) and after completion of induction chemotherapy (day-28). Fifteen non-malignant cases were recruited as controls. Methods WT-1 gene expression and methylation status were assessed during both intervals (day-0 and day-28) by performing real-time polymerase chain reaction (RT-PCR) and methylation-specific polymerase chain reaction (MS-PCR). A flow cytometric immune-phenotyping study was also performed using a panel of monoclonal antibodies specific to AML. Results Of the 143 subjects studied, 123 (83.67%) patients showed overexpression of the WT-1 gene at the time of diagnosis as compared with patients in complete remission (CR) remission or control samples (P=<0.001). Moreover, robust hypermethylation of the WT1 promoter was observed in 103 (70.06%) AML patients at the time of diagnosis as compared with patients in complete remission (CR) remission or control samples (P=<0.001). The flow cytometric immune-phenotyping study demonstrated that CD34, CD45, CD117, CD38, CD13, CD33, and CD56 were expressed as positive in most cases, while cCD79a CD19 CD7, cCD3, CD16, CD123, CD11b were negatively observed. In addition, significant positive correlations between WT-1 expression and methylation levels on day-0 and day-28 were observed (P<0.001). Conclusions Overexpression and hypermethylation of the WT-1 gene correlate with the leukemic burden in most cases of AML. Thus, this gene can be considered a promising molecular marker for early diagnosis and MRD detection and a target for developing novel therapeutic approaches against AML.