signature=f1fc1773ddf79d526ebf5741013ca280,Transcriptomic Profile Identified a Specific Signature in...

摘要：

Background. Acute Myeloid Leukemia is a heterogeneous disease, characterized by the uncontrolled proliferation of hematopoietic precursor cells. AML accounts for 15% of acute leukemias in children, with a cure rate of 55-60% as overall survival (Pession A Blood 2013). Complete remission (CR) is achieved in 80-85% of children with AML. Current protocol defines CR by morphological evaluation (blasts 60%); in the other three cases we detected, by flow-cytometry, a blast rate between 10-20%. In the attempt to compare these cases with those presenting with very high-risk (VHR) features but achieving CR after induction (Primary Induction Responder, PIR), we included in our analyses 8 children with VHR-PIR-AML, diagnosed and treated in our Center: 2 with FLT3-ITD and NUP98-NSD1 fusion genes; 2 with FLT3-ITD and complex karyotype; 2 with DEK-CAN fusion genes (1 with FLT3-ITD); 2 with NUP98 rearrangements and complex karyotype. We performed whole transcriptome analyses using Human Transcriptome Array 2.0 (Affymetrix). Then we compared our analyses with a recently published data set, including a large population of children with AML (Bisio V Leukemia 2017). Based on transcriptomic findings, we determined the expression of specific genes, by Real-time PCR using gene-specific probes (UPL probes by Roche Merck, Germany), applying a QuantStudio7 Flex System technology (Applied Biosystem, California, USA) and the relative quantification method (the comparative 2-ΔΔCt method). We applied a t-student test, for statistical analyses among different subgroups. Results. A direct comparison of PIF-AML vs PIR-AML subgroups revealed 919 protein-coding and 8 pri-miRNA overexpressed genes (fold-change >3, FDR adjusted p value -3, FDR adjusted p value <0.05). Marked differences were observed in cancer-related genes such as KIT, PTEN, FLT3 and POU4F1, and in cell-cycle related genes such as cyclin transcripts (CNNG1, CCNA1, CCND2, CCNJ, CCNC, CCNY). Moreover some genes that discriminate PIF from PIR subgroups are also differentially expressed between cases with NUP98-negative and NUP98-positive AML (PDGFD, CCNA1). The latter group is considered as a VHR-AML subtype. Based on these findings, we selected several genes including HOX-B3, HOX-B5, CEBP-D, CCNA1, PDGF-D and performed a Real-Time PCR with specific-gene probes. Comparing gene expression data of our 4 PIF-AML cases with 8 VHR-PIR-AML cases, we observed a strong statistically significant different expression in CCNA1 (p<0.0001) and PDGF-D (p<0.0001). Conclusions. Our findings defined a characteristic profile of PIF-AML, highlighting the overexpression of cell-cycle related genes (CCNG1, CCNA1) and several potential targets (KIT, FLT3, PDGF-D) to be considered for an alternative treatment in this very poor-prognostic subgroup of patients. In conclusion our results suggest that PIF-AML samples bear a specific gene expression profile, not overlapping with those of known molecular and karyotype AML subgroups. However, a larger population study is needed to confirm our preliminary data. Disclosures No relevant conflicts of interest to declare.

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