Background: Acute myelogenous leukemia (AML) is consideredas one of the most common types of hemato-oncologydeficiencies in adults. Despite the use of aggressive chemotherapy,relapse after remission and refractoriness to inductionchemotherapy still remain as the most common therapeuticfailures in AML. It seems that a rare population of cells calledLeukemia stem cells (LSCs), are responsible for initiating therapyresistance and relapse.
Gene Regulatory Network (GRN)is an adequate bioinformatics tool used in mapping gene interactionsand also, pinpointing key genes which may regulatethe stemness core in LSC.Therefore, in this study we appliedGRN to identify a precise list of potential biomarkers and targetgenes, which may play an effective role in targeting these cells.Materials and Methods: This study was conducted in twophases, ۱) in silico ۲) in vitro. In the in silico phase our aimwas to identify key genes and their related signaling pathwayswhich may be regulating the stemness core in LSCs. In thisphase, we obtained a data set of ۴۹۵ Gene expression profiles(GSE۷۶۰۰۹). Differentially expression analysis was performedbetween two defined groups of LSC+ and LSC- using limmapackage in R Software (۳.۴.۲ version). In addition, networkconstruction was conducted using ARACNE algorithm and cytoscapesoftware. Subsequently, ۲۴ “Hub genes”, genes whichhave the most interaction with other genes, were identified andpathway analysis was conducted using different tools in differentdatabases. Hence, based on our selection criteria, the listof genes was narrowed down to ۴ genes, to be evaluated in thewet-lab phase. In the in vitro phase, two approaches were followed.First, we used two small molecules SANT-۱ and RG۱۰۸to inhibit two important signaling pathways in KG-۱ cell line.In the second step, KG-۱ cell line was transfected with a combinationof four siRNAs to evaluate stemness decrease. In the insilico phase we determined ۴ genes and their related signalingpathways. Moreover, we indicated that by using the combinationof SANT-۱ and RG۱۰۸, a significant decrease in stemnessis observed. We also observed that, the combinational inhibitionof ۳ siRNAs had a significant role in decreasing stemness.By comparing the two inhibitory methods, siRNA transfectionexhibited more effective results. Results: We determined a candidate set of genes that are significantlyrelated to stemness and self renewal pathways in AML.Conclusion: Our data suggested that by using this bioinformaticsapproach, we were able to attain a list of candidate geneswhich may have a significant role in regulating the stemnesscore in LSCs.