Genes In Acute Myeloid Leukemia Patients Research Articles

Unveiling mitochondrial and ribosomal gene deregulation and tumor microenvironment dynamics in acute myeloid leukemia.

Acute myeloid leukemia (AML) is a malignant clonal hematopoietic disease with a poor prognosis. Understanding the interaction between leukemic cells and the tumor microenvironment (TME) can help predict the prognosis of leukemia and guide its treatment. Re-analyzing the scRNA-seq data from the CSC and G20 cohorts, using a Python-based pipeline including machine-learning-based scVI-tools, recapitulated the distinct hierarchical structure within the samples of AML patients. Weighted correlation network analysis (WGCNA) was conducted to construct a weighted gene co-expression network and to identify gene modules primarily focusing on hematopoietic stem cells (HSCs), multipotent progenitors (MPPs), and natural killer (NK) cells. The analysis revealed significant deregulation in gene modules associated with aerobic respiration and ribosomal/cytoplasmic translation. Cell-cell communications were elucidated by the CellChat package, revealing an imbalance of activating and inhibitory immune signaling pathways. Interception of genes upregulated in leukemic HSCs & MPPs as well as in NKG2A-high NK cells was used to construct prognostic models. Normal Cox and artificial neural network models based on 10 genes were developed. The study reveals the deregulation of mitochondrial and ribosomal genes in AML patients and suggests the co-occurrence of stimulatory and inhibitory factors in the AML TME.

SET-PP2A complex as a new therapeutic target in KMT2A (MLL) rearranged AML

KMT2A-rearranged (KMT2A-R) is an aggressive and chemo-refractory acute leukemia which mostly affects children. Transcriptomics-based characterization and chemical interrogation identified kinases as key drivers of survival and drug resistance in KMT2A-R leukemia. In contrast, the contribution and regulation of phosphatases is unknown. In this study we uncover the essential role and underlying mechanisms of SET, the endogenous inhibitor of Ser/Thr phosphatase PP2A, in KMT2A-R-leukemia. Investigation of SET expression in acute myeloid leukemia (AML) samples demonstrated that SET is overexpressed, and elevated expression of SET is correlated with poor prognosis and with the expression of MEIS and HOXA genes in AML patients. Silencing SET specifically abolished the clonogenic ability of KMT2A-R leukemic cells and the transcription of KMT2A targets genes HOXA9 and HOXA10. Subsequent mechanistic investigations showed that SET interacts with both KMT2A wild type and fusion proteins, and it is recruited to the HOXA10 promoter. Pharmacological inhibition of SET by FTY720 disrupted SET-PP2A interaction leading to cell cycle arrest and increased sensitivity to chemotherapy in KMT2A-R-leukemic models. Phospho-proteomic analyses revealed that FTY720 reduced the activity of kinases regulated by PP2A, including ERK1, GSK3β, AURB and PLK1 and led to suppression of MYC, supporting the hypothesis of a feedback loop among PP2A, AURB, PLK1, MYC, and SET. Our findings illustrate that SET is a novel player in KMT2A-R leukemia and they provide evidence that SET antagonism could serve as a novel strategy to treat this aggressive leukemia.

Investigating the interplay of hematological parameters, CD markers, genetic polymorphisms, and database mutations in the IL15 gene in acute myeloid leukemia patients.

Acute myeloid leukemia (AML) is a heterogeneous malignancy characterized by the clonal expansion of myeloid precursor cells in the bone marrow. In this study, we investigated the interplay of hematological parameters, CD markers, genetic polymorphisms, and database mutations in the interleukin 15 (IL15) gene in AML patients. We enrolled 59 newly diagnosed AML patients and analyzed their bone marrow specimens using flow cytometry and molecular techniques. The hematological parameters of the AML patients revealed a significant increase in platelet count and RBC, Hb, and HCT levels compared to healthy individuals. CD marker expression analysis revealed upregulation of CD33, CD45, CD13, CD117, CD38, HLA-DR, CD15, CD64, MPO, CD34, and CD11c in AML patients. Molecular analysis showed 15 mutations in different positions of exon 8 of the IL15 gene, with the most frequent mutation being a homozygous mutation resulting from a nucleotide substitution. Additionally, 10 novel heterozygous mutations were identified in different locations of chromosome 4, with a low variant rate. Finally, database analysis of gnomAD and Mutagene revealed a high number of potential driver mutations in the IL15 gene in leukemia patients. These results provide valuable insights into the genetic and immunophenotypic characteristics of AML patients and highlight the potential role of IL15 in AML pathogenesis.

Molecular and clinical significance of FLT3, NPM1, DNMT3A and TP53 mutations in acute myeloid leukemia patients.

Acute myeloid leukemia (AML) is a type of blood cancer that affects the bone marrow and blood cells. AML is characterized by the rapid growth and accumulation of abnormal white blood cells, known as myeloblasts, which interfere with the production of normal blood cells. The main aim was to determine the relationship between these genetic alterations and the clinico-haematological parameters and prognostic factors with therapy for Iraqi patients with AML. We used Sanger Sequencing to detect the mutations in 76 AML patients. Clinical data of AML patients were retrospectively analysed to compare the prognosis of each gene mutation group. Somatic mutations were identified in 47.4% of the enrolled patients in a core set of pathogenic genes, including FLT3 (18 patients, 23.7%), DNMT3A (14, 18.4%), NPM1 (11, 14.5%) and TP53 (5, 6.8%). As multiple mutations frequently coexisted in the same patient, we classified patients into 10 further groups. Two novel mutations were detected in FLT3-ITD, with new accession numbers deposited into NCBI GenBank (OP807465 and OP807466). These two novel mutations were computationally analysed and predicted as disease-causing mutations. We found significant differences between patients with and without the detected mutations in disease progression after induction therapy (remission, failure and death; pv = < 0.001) and statistically significant differences were reported in total leukocyte count (pv = < 0.0001). These genes are among the most frequently mutated genes in AML patients. Understanding the molecular and clinical significance of these mutations is important for guiding treatment decisions and predicting patient outcomes.

The Class IIA Histone Deacetylase (HDAC) Inhibitor TMP269 Downregulates Ribosomal Proteins and Has Anti-Proliferative and Pro-Apoptotic Effects on AML Cells.

Acute myeloid leukemia (AML) is a hematopoietic malignancy characterized by altered myeloid progenitor cell proliferation and differentiation. As in many other cancers, epigenetic transcriptional repressors such as histone deacetylases (HDACs) are dysregulated in AML. Here, we investigated (1) HDAC gene expression in AML patients and in different AML cell lines and (2) the effect of treating AML cells with the specific class IIA HDAC inhibitor TMP269, by applying proteomic and comparative bioinformatic analyses. We also analyzed cell proliferation, apoptosis, and the cell-killing capacities of TMP269 in combination with venetoclax compared to azacitidine plus venetoclax, by flow cytometry. Our results demonstrate significantly overexpressed class I and class II HDAC genes in AML patients, a phenotype which is conserved in AML cell lines. In AML MOLM-13 cells, TMP269 treatment downregulated a set of ribosomal proteins which are overexpressed in AML patients at the transcriptional level. TMP269 showed anti-proliferative effects and induced additive apoptotic effects in combination with venetoclax. We conclude that TMP269 exerts anti-leukemic activity when combined with venetoclax and has potential as a therapeutic drug in AML.

The study of long noncoding RNA SNHG5 and PANDAR genes expression in newly diagnosed egyptian adult acute myeloid leukemia patients

BACKGROUND: Due to their impact on crucial steps in hematopoiesis, long noncoding RNAs (lncRNAs) deregulation potentially accelerates the growth and development of blood cancers like acute myeloid leukemia (AML). The study aimed to look into different expression patterns, prognostic value, and clinical importance of lncRNA small nucleolar RNA host gene 5 (SNHG5) and promoter of cyclin-dependent kinase inhibitor 1A antisense DNA damage-activated RNA (PANDAR) genes in Egyptian adult patients with AML. SUBJECTS AND METHODS: The case–control study was conducted between 2019 and 2022 at the Clinical Pathology Department at the National Cancer Institute, Cairo University, Egypt. The study involved 80 recently diagnosed patients with AML and 20 healthy controls. Real-time quantitative reverse transcription polymerase chain reaction was used to assess the levels of expression of SNHG5 and PANDAR genes. RESULTS: In comparison to healthy controls, there was a significantly higher SNHG5 gene expression (P = 0.026) and PANDAR expression (P &lt; 0.001) in patients’ bone marrow samples. The study of the correlations revealed a significant positive association between SNHG5 and PANDAR genes in AML patients. The overall survival (OS) was significantly better in the low SNHG5 gene expression group than in the high SNHG5 gene expression group. No significant difference was detected regarding the disease-free survival (DFS) between patients with low expression and high expression of the SNHG5 gene. No significant variation between high PANDAR gene and low PANDAR gene expression regarding OS and DFS. CONCLUSION: SNHG5 and PANDAR may have a pathogenic role in AML, and their overexpression might be considered a marker for diagnosis in AML patients in Egypt. SNHG5 expression can be used as a predictor for OS, while PANDAR expression cannot be used as a predictor for OS or DFS in patients.

Expression Changes of SIRT1 and FOXO3a Significantly Correlate with Oxidative Stress Resistance Genes in AML Patients.

The increased metabolism in acute myeloid leukemia (AML) malignant cells resulted in the production of high levels of free radicals, called oxidative stress conditions. To avoid this situation, malignant cells produce a considerable amount of antioxidant agents, which will lead to the release of a continuous low level of reactive oxygen species (ROS), causing genomic damage and subsequent clonal evolution. SIRT1 has a key role in driving the adaptation to this condition, mainly through the deacetylation of FOXO3a that affects the expression of oxidative stress resistance target genes such as Catalase and Manganese superoxide dismutase (MnSOD). The aim of this study is to simultaneously investigate the expression of SIRT1, FOXO3a, and free radical-neutralizing enzymes such as Catalase and MnSOD in AML patients and measure their simultaneous change in relation to each other. The gene expression was analyzed using Real Time-PCR in 65 AML patients and 10 healthy controls. Our finding revealed that expression of SIRT1, FOXO3a, MnSOD and Catalase was significantly higher in AML patients in comparison to healthy controls. Also, there was a significant correlation between the expression of SIRT1 and FOXO3a, as well as among the expression of FOXO3a, MnSOD and Catalase genes in patients. According to the results, the expression of genes involved in oxidative stress resistance was higher in AML patients, which possibly contributed to the development of malignant clones. Also, the correlation between the expression of SIRT1 and FOXO3a gene reflects the importance of these two genes in increased oxidative stress resistance of cancer cells.

Fatty acid metabolism predicts prognosis and NK cell immunosurveillance of acute myeloid leukemia patients

Cell metabolic reprogramming is a hallmark of tumor prognosis, and fatty acid metabolism (FAM) plays a crucial role in the tumor microenvironment (TME). However, the relationship between FAM, TME, and prognosis of acute myeloid leukemia (AML) patients remains elusive. We extracted the single-cell RNA sequencing (scRNA-Seq) and bulk transcriptome data of AML patients from the TCGA and GEO databases and assessed the relationship between FAM, TME, and AML patient prognosis. We also performed functional enrichment (FE) assay to evaluate the significance of FAM in anti-AML immunosurveillance. Our scRNA-Seq analysis revealed that the leukemic stem cell (LSC)-enriched population exhibited elevated levels of FAM-related genes. Using these FAM-related genes, we developed a prognostic model that accurately estimated AML patient outcome. FE analysis showed that FAM was strongly related to alterations of TME-based immunosurveillance in AML patients. More importantly, we demonstrated that FAM inhibition via pharmaceutical targeting of PLA2G4A, a highly expressed FAM gene in AML patients with poor prognosis, enhanced the NK cell-mediated immunosurveillance in leukemia cells. Leukemic stem cell (LSC)-enriched population exhibited elevated levels of FAM-related genes. We have successfully established the FAM formula that predicts AML patient prognosis and alterations in the TME-based immunosurveillance. We also found that PLA2G4A was a highly expressed FAM gene in AML patients with poor prognoses. Pharmaceutical targeting of PLA2G4A increased the expression of NKG2DL in leukemia cells in vitro and thus enhanced the NK cell-mediated immunosurveillance.

Clinical Significance of EZH2 in Acute Myeloid Leukemia.

In this paper, we have focused on the investigation of the expression level of the enhancer of zeste homolog 2 (EZH2) gene in bone marrow mononuclear cells of acute myeloid leukemia (AML) patients and analyze the relationship between EZH2 gene expression and EMI. The expression of EZH2mRNA in bone marrow mononuclear cells of 26 patients with incipient AML was detected by qRT-PCR, and the relationship between EZH2mRNA expression and clinical characteristics was analyzed. EZH2 mRNA expression was increased in 26 AML patients. EZH2 gene expression in male patients was significantly higher than that in female patients. The expression of EZH2 in the group with extramedullary infiltration (EMI) was significantly higher than that in the group without EMI. The patients were divided into different groups according to the chromosomal karyotype and prognosis. Statistical analysis showed that the expression level of the medium-risk group was significantly higher than that of the low-risk group, while there was no statistical difference in other groups (P > 0.05). The expression of EZH2 gene in AML patients was closely related to EMI, and the expression of EZH2 in AML cells was closely related to cell migration ability. EZH2 is expected to be one of the indicators of disease recurrence.

The signature based on seven genomic instability-related genes could predict the prognosis of acute myeloid leukemia patients

ABSTRACT Background Acute myeloid leukemia (AML) is the most common acute blood malignancy in adults. The complicated and dynamic genomic instability (GI) is the most prominent feature of AML. Our study aimed to explore the prognostic value of GI-related genes in AML patients. Methods The mRNA data and mutation data were downloaded from the TCGA and GEO databases. Differential expression analyses were completed in limma package. GO and KEGG functional enrichment was conducted using clusterProfiler function of R. Univariate Cox and LASSO Cox regression analyses were performed to screen key genes for Risk score model construction. Nomogram was built with rms package. Results We identified 114 DEGs between high TMB patients and low TMB AML patients, which were significantly enriched in 429 GO terms and 13 KEGG pathways. Based on the univariate Cox and LASSO Cox regression analyses, seven optimal genes were finally applied for Risk score model construction, including SELE, LGALS1, ITGAX, TMEM200A, SLC25A21, S100A4 and CRIP1. The Risk score could reliably predict the prognosis of AML patients. Age and Risk score were both independent prognostic indicators for AML, and the Nomogram based on them could also reliably predict the OS of AML patients. Conclusions A prognostic signature based on seven GI-related genes and a predictive Nomogram for AML patients are finally successfully constructed.

The Prognostic Value of FOSB Gene in Acute Myeloid Leukemia

The genome, transcriptome, gene chip and clinical information from multiple public databases were statistical analyzed. The expression of FOSB gene in AML patients was significantly higher than that in normal people. The prognostic analysis of the 163 patients showed that the patients with high FOSB expression showed longer OS and EFS than those with FOSB low expression. The patients were further divided into chemotherapy group and allogeneic hematopoietic stem cell transplantation (allo-HSCT) group according to the treatment method, and then each group was divided into two subgroups (FOSBhigh, FOSBlow) according to the median expression level of FOSB. In the allo-HSCT group, the patients with FOSB high expression was longer event-free survival (EFS: P=0.017) and overall survival (OS: P=0029). At the same time, allo-HSCT in patients with high FOSB expression could improve the prognosis of the patients (Chemotherapy vs Allo-HSCT, OS: P<0.001, EFS: P=0.007). Multivariate analysis showed that the high expression of FOSB was an independent favorable prognostic factor for EFS and OS (EFS: HR=0.501， P=0.019; OS: HR=0.461， P=0.009) of the patients. The high expression of FOSB indicated a good prognosis for acute myeloid leukemia.

Altered expression of NEAT1 variants and P53, PTEN, and BCL-2 genes in patients with acute myeloid leukemia

BackgroundNewly discovered evidence showed that long non-coding RNAs (lncRNAs) can play crucial roles in the development of cancer therapies. Nuclear Enriched Abundant Transcripts 1 (NEAT1) is one of the lncRNAs the expression of which changes in different cancers. The role of NEAT1 in apoptosis is discussed in various malignancies. This study aimed to determine the NEAT1 expression and its correlation with P53, PTEN, and BCL-2 genes expression in acute myeloid leukemia (AML) patients. MethodIn this study, using quantitative real-time polymerase chain reaction (qRT-PCR), we analyzed the expression of NEAT1, P53, PTEN, and BCL-2 genes in 21 AML patients. Moreover, relative quantification analysis was performed by delta-delta CT method (2 −ΔΔCT) method. ResultsOur results showed that NEAT1 expression was significantly lower in AML patients compared to healthy controls (P-value < 0.05). While a significant correlation was observed between the expression levels of NEAT1 and PTEN genes in AML patients (P-value < 0.05), there was no correlation between the expression levels of NEAT1 with P53 and BCL-2 (P-value > 0.05). ConclusionAccording to the results, increased expression of NEAT1 gene may play a role in the apoptosis of AML cells, despite the oncogenic role in most solid tumors.

Identification and Validation of a Prognostic Risk-Scoring Model Based on Ferroptosis-Associated Cluster in Acute Myeloid Leukemia.

Background: Emerging evidence has proven that ferroptosis plays an important role in the development of acute myeloid leukemia (AML), whereas the exact role of ferroptosis-associated genes in AML patients’ prognosis remained unclear. Materials and Methods: Gene expression profiles and corresponding clinical information of AML cases were obtained from the TCGA (TCGA-LAML), GEO (GSE71014), and TARGET databases (TARGET-AML). Patients in the TCGA cohort were well-grouped into two clusters based on ferroptosis-related genes, and differentially expressed genes were screened between the two clusters. Univariate Cox and LASSO regression analyses were applied to select prognosis-related genes for the construction of a prognostic risk-scoring model. Survival analysis was analyzed by Kaplan–Meier and receiver operator characteristic curves. Furthermore, we explored the correlation of the prognostic risk-scoring model with immune infiltration and chemotherapy response. Risk gene expression level was detected by quantitative reverse transcription polymerase chain reaction. Results: Eighteen signature genes, including ZSCAN4, ASTN1, CCL23, DLL3, EFNB3, FAM155B, FOXL1, HMX2, HRASLS, LGALS1, LHX6, MXRA5, PCDHB12, PRINS, TMEM56, TWIST1, ZFPM2, and ZNF560, were developed to construct a prognostic risk-scoring model. AML patients could be grouped into high- and low-risk groups, and low-risk patients showed better survival than high-risk patients. Area under the curve values of 1, 3, and 5 years were 0.81, 0.827, and 0.786 in the training set, respectively, indicating a good predictive efficacy. In addition, age and risk score were the independent prognostic factors after univariate and multivariate Cox regression analyses. A nomogram containing clinical factors and prognostic risk-scoring model was constructed to better estimate individual survival. Further analyses demonstrated that risk score was associated with the immune infiltration and response to chemotherapy. Our experiment data revealed that LGALS1 and TMEM56 showed notably decreased expression in AML samples than that of the normal samples. Conclusion: Our study shows that the prognostic risk-scoring model and key risk gene may provide potential prognostic biomarkers and therapeutic option for AML patients.

Identification of Mutations in GATA2 Gene in Adult Bengali Bangladeshi Acute Myeloid Leukemia (AML) Patients

AML is a malignant disease of the bone marrow in which hematopoietic precursors are arrested in an early stage of development due to genetic alterations in normal hematopoietic stem cells. Several genes are identified for the disease and GATA2 gene is one of the important genes among them because mutations of GATA2 gene suppress leukemogenesis. In a country like Bangladesh, genetic services are at an early stage of development or even not yet developed. There is no adequate reporting on GATA2 gene mutations in adult Bengali Bangladeshi AML patients. The aim of the study was to determine the mutations of GATA2 gene in AML patients in the adult Bengali Bangladeshi population. The present study was a cross-sectional type of descriptive study. The study was carried out in the Genetic Research Center (GRC) of the Department of Anatomy, Bangabandhu Sheikh Mujib Medical University (BSMMU), Bangladesh during the period of January 2018 to December 2018. The study includes 50 adult Bengali Bangladeshi AML patients. Informed written consent was obtained from the participants. For identifying the mutations peripheral venous blood was obtained from each patient and sequencing was done using ABI Sanger Sequencer 3730. Research shows the mean age is 25±14.2 and average age group of adult AML patients of Bengali Bangladeshis is 18-29 years (54%), among them 31 (62%) and 19 (38%) were females and males respectively. GATA2 gene mutations were identified in 1 of the 50 AML patients and shows synonymous, frameshift and missense variant compared with the database of dbSNP. Identification of different variant with this small sample size emphasizes the importance of exploring the genetic makeup of adult Bengali Bangladeshi AML patients to develop a database for proper screening and genetic counseling of the disease.

ARHGAP6 EXPRESSION IS ASSOCIATED WITH MOLECULAR RISK AND IMPACTS CLINICAL OUTCOMES IN ACUTE MYELOID LEUKEMIA

Despite the advances in the large-scale generation of genomic data from acute myeloid leukemia (AML) patients, much effort is still needed to convert these data into biological functions, which may pave avenues for the discovery of new prognostic markers and therapeutic targets. In a previous study, four cytoskeleton-related genes (ARHGAP6, EZR, MAP3K11 and PAK1) were identified as a prognostic factor in AML. In the present study, we detail the prognostic impact, as well as its functional genomic associations, of the ARHGAP6 gene in AML patients. Clinical and genomic data were obtained from the TCGA AML cohort (n = 173). The clinical impact (overall survival [OS] and disease-free survival [DFS]) for ARHGAP6 expression was investigated by the proportional hazards Cox regression model. Functional genomic analyzes were performed by gene set enrichment analysis (GSEA) and Gene Ontology (GO) from differentially expressed (DEG) genes obtained from the Galaxy tool. All p values were two sided with a significance level of 5%. ARGHAP6 expression was higher in the intermediate and high molecular risk groups in AML patients (all p < 0.01). High ARGHAP6 expression was identified as an independent risk factor for OS (HR = 1.92; p = 0.001) and DFS (HR = 1.96; p = 0.01) when analyzed with confounders, age, gender, white blood cell count, and molecular risk. In the GSEA, a total of 30 cellular and molecular events were found to be positively enriched in high ARHGAP6-expressing AML patients, which we highlighted the signaling pathways: IL6/JAK2/STAT3 (NES: 1.62; p < 0.001), NOTCH (NES: 1.60; p < 0.001), KRAS (NES: 1.59; p < 0.001), IL2/STAT5 (NES: 1.51; p < 0.001), TGF BETA (NES: 1.39; p = 0.03) and BETA CATENIN SIGNALING (NES: 1.36; p = 0.04). In GO analyses, high ARHGAP6 expression was associated with important processes for cancer development and progression such as cell signaling mediated by surface receptors, signal transduction, linkage to growth factors, and regulation of cell population proliferation (all p < 0.001). ARHGAP6 is a RhoGAP that acts converting RhoGTPases in its inactive GDP-bound state, which may regulate the interactions of signaling molecules with the actin cytoskeleton. It promotes continuous elongation of cytoplasmic processes during cell motility and simultaneous retraction of the cell body changing the cell morphology. The role of ARHGAP6 is still poorly established in cancer, but it is known to modulate pathways of RhoGTPases. In this sense, our study is a pioneer in the identification of this gene in processes related to neoplastic hematopoiesis, which will guide future investigations. ARHGAP6 gene expression was identified as an independent marker of poor prognosis in AML patients. The GSEA suggested that high ARHGAP6 expression is associated with activation of relevant processes to the malignant phenotype of AML. FAPESP, CNPq and CAPES.

Transcription analysis of a histones modifiers panel coupled with critical tumor suppressor genes displayed frequent changes in patients with AML.: mRNA levels of histones modifiers and TSGs in AML

Epigenetic alterations could cause leukemia through the activation of normally silent loci or silencing of normally active loci. We herein aimed to compare the expression patterns of a histone modifiers panel consisted of SUV39H1, PRDM16, UHRF2, KDM2B, and KDM3C between acute myeloid leukemia(AML) cells and normal cells and to assess the correlation of these genes with the expression of vital tumor suppressor genes, including p16INK4A and p53.Bone marrow or peripheral blood samples of 50 AML patients at diagnosis and also 18 subjects with a normal hematopoietic system as a control group were obtained after informed consent. Then, qRT-PCR was performed to determine the expression levels of the aforementioned genes. We found a broad alteration in the expression signature of five out of seven studied genes in AML patients as compared with the control group. UHRF2 and p53 were remarkably downregulated in AML patients (P<0.001), while SUV39H1, PRDM16, and KDM3C were significantly overexpressed (P<0.01). Based on the Spearman rank correlation, SUV39H1 and KDM2B negatively regulated both p16INK4A and p53 expression. Taken together, our findings provided preliminary evidence regarding the pervasive mRNA perturbation of histone modifiers and their plausible influences on critical tumor suppressor genes. Future studies in this area would be required to assist in establishing these results in the clinical practice of AML patients.

Comprehensive Expression and Prognosis Analyses of ITGB1 in AML

Background: Acute myeloid leukemia (AML) is a dangerous type of leukemia. The emergence of multidrug resistance (MDR) and recurrence limits the prognosis and survival of patients. In recent studies, we have known that the bone marrow microenvironment was closely related to the poor prognosis of AML. However, the underlying mechanisms are still far from fully understood. By utilizing the bioinformatics analysis, we screened out integrin B1 (ITGB1) as the hub gene, which is associated with the bone marrow microenvironment mediated changes of AML cells, with expression profile GSE73157 downloaded from the National Center for Biotechnology Information-Gene Expression Omnibus (NCBI-GEO) database. Methods: R studio software was used to screen out candidate hub genes and further visualize the differential expression. R package ‘limma’ was to find out differentially expressed genes (DEGs). Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were conducted by R package “clusterProfiler”. Furthermore, the protein-protein interaction (PPI) network was also performed by the online tool STRING and software Cytoscape. Last but not least, online tools PrognoScan and GEPIA were utilized for the evaluation of the clinical significance of the selected hub gene. P and Cox p-value &lt;0.05 was considered to be statistically significant. Results: ITGB1 was filtrated as the only hub gene in this profile. We found that patients with high expression of ITGB1 had significantly longer overall survival (OS) than those with low expression (COX p-value= 0.016730). Besides, the expression of the ITGB1 gene in AML patients is lower than that in normal people significantly (p-value&lt;0.01). Conclusion: We identified ITGB1 as a key gene in the bone marrow microenvironment mediated poor prognosis in AML. The down-regulated expression of ITGB1 was related to AML patients’ poor outcomes. ITGB1 may be a potential marker for predicting and guiding AML treatment.

A Three-Genes Signature of Anti-Apoptotic Genes Showed Different Mechanisms in Preventing Apoptosis in Acute Myeloid Leukemia Cells and Could Provide an Useful Model to Establish the Proper BH3-Mimetich Drug or Combination

BACKGROUNDSeveral biological mechanisms sustain leukemic development, but the inhibition of apoptotic death, which should be induced by the genetic instability of the disease, is surely one the key neoplastic alterations. Apoptotic regulation involves complex pathways of positive and negative regulators which may and do represent a target for personalized therapy. Furthermore, the overexpression of specific anti-apoptotic genes likely represent a useful prognostic instrument. Nowadays, BCL-2 inhibitors have successfully passed preliminary phases of clinical experimentation, however the involvement of other essential genes, such as MCL-1 and BCL2L1 (coding for bcl-xL), has been proposed as resistance mechanism to BCL-2 inhibitors and as novel therapeutic target in Acute Myeloid Leukemia (AML).Our study aims to investigate the expression of anti-apoptotic genes in AML patients to cluster them in different categories and study their impact on prognosis, in order to reveal possible mechanism of resistance to BCL-2 inhibitors and stratify patients that are more likely to take advantage of administration or co-administration of one between MCL-1 and BCL-2/BCL2L2 inhibitors. METHODSWe performed Human Transcriptome Array 2.0 (Affymetrix) in a cohort of 59 new diagnosed AML. Gene Expression Profiling (GEP) analysis has been conducted with Transcriptome Array V3.0 Software (Affymetrix). K-means clustering has been used to categorize different cluster of patients basing on BCL-2, MCL-1 and BCL2L1 gene expression. A pool of 7 healthy donor was used to normalize expression data. Survival analysis was conducted with Kaplan-Meyer method and differences in survival were assessed with Log-Rank test. RESULTSAnalyzing BCL-2, MCL-1 and BCL2L1 gene expression values in 59 AML patients at diagnosis we established 5 categories where patients are almost uniformly assigned (Figure 1a): 11/59 patients in Cluster 1, characterized by the only one overexpression of MCL1; 27/59 patients in Cluster 2, which presents low expression of all analyzed genes; 2/59 patients in Cluster 3, where BCL2L1 is strongly overexpressed; 6/59 patients in Cluster 4, distinguished by BCL-2 overexpression and 13/59 patients in Cluster 5, which high values of expression of all 3 genes. Thus, Clusters 2 and 5, where no gene is overexpressed in comparison with the others, include the majority of patients (40/59, approximately 68%), suggesting that apoptotic death control is realized thanks to an intricate debate between these 3 considered proteins and BCL-2 should not be considered the unique oncogenic anti-apoptotic mechanism. Furthermore, the 3-D Scatter Plot (Figure 1A) allows us to observe how the patients out of the central cloud (Clusters 2-5) always present one of the three anti-apoptotic genes overexpressed, and when BCL-2 level is progressively low (Cluster 1-2-3), MCL-1 and BCL2L1 values are high, respectively. Therefore, this documented behavior may indicate the presence of many ways used by leukemic cell to prevent apoptosis, with one of them almost always activated.In term of Overall Survival (OS), unsurprisingly no statistically significant difference arised between Cluster 1-3-4 where one different anti-apoptotic gene is overexpressed (Figure 1b), underling the probable equal effect of each mechanism in inhibiting apoptotic death and in sustaining leukemic progression. Finally, analyzing the impact only of BCL-2 expression on OS in a cohort of 36 young patients treated with chemotherapy it is possible to observe a trend close to statistically significance (p=0.07)(Figure 1c), unreached probably because of the relative limitation of the sample, confirming the BCL-2 induced chemoresistance. CONCLUSIONSApoptotic death regulation is precisely controlled equally by BCL-2, MCL-1 and BCL2L2, their expression is quite balanced in AML patients evaluated. One out of these three genes is almost always overexpressed, reaffirming the key role of anti-apoptotic oncogenes in leukemogenesis. Even though BCL-2 expression influence patients prognosis, MCL-1 and BCL2L2 overexpression has been documented when BCL-2 is normally or underexpressed. In conclusion, BCL-2 inhibitors resistance may be caused by MCL-1 and BCL2L2 overexpression detected in patients BCL-2 independents. Moreover, a synthetic lethality approach should be realized to switch-off leukemic anti-apoptotic multiple mechanisms. [Display omitted] DisclosuresMartinelli:Celgene: Consultancy; Amgen: Consultancy; Johnson&Johnson: Consultancy; Ariad/Incyte: Consultancy; Pfizer: Consultancy; Roche: Consultancy.

Identification of RHOBTB2 aberration as an independent prognostic indicator in acute myeloid leukemia.

Rho-related BTB domain (RhoBTB) proteins belong to Rho guanosine triphosphatases (GTPases). Their putative role implicated in carcinogenesis has been supported by accumulating evidence. However, their expression pattern and potential role in acute myeloid leukemia (AML) remain unclear.We profiled RHOBTB mRNA expression via the Gene Expression Profiling Interactive Analysis 2 (GEPIA2) database. Survival analysis was conducted with GEPIA2 and UALCAN. Univariate and multivariate Cox regression analyses were performed to validate RHOBTB genes as independent prognostic indicators in the LAML cohort from The Cancer Genome Atlas (TCGA). Data regarding expression in different subtypes and relationships with common disease-related genes were retrieved from UALCAN. Co-expressed genes were screened out and subsequently subjected to functional enrichment analysis.We observed aberrant transcription levels of RHOBTB genes in AML patients. RHOBTB2 was identified as a prognostic candidate for overall survival (OS), independent of prognosis-related clinical factors and genetic abnormalities. Moreover, RHOBTB2 expression was increased in non-acute promyelocytic leukemia (APL) subtypes, patients without FLT3 mutation and PML/RAR fusion, and imparted a positive correlation with the expression of FLT3, FHL1, and RUNXs. Co-expressed genes of RHOBTB2 were enriched in functional pathways in AML.Our findings suggest that RHOBTB2 might be a novel biomarker and independent prognostic indicator in AML and provide insights into the leukemogenesis and molecular network of AML.

The Expression of WTAP Gene in Acute Myeloid Leukemia and Its Clinical Significance

To investigate the expression of WTAP gene in acute myeloid leukemia (AML) and its clinical significance. 74 acute myeloid leukemia patients with non-M3 type and 19 normal donors were selected, and real-time quantitative polymerase chain reaction was used to detect the mRNA expression level of WTAP gene in their bone marrow cells. The relationship between the mRNA expression level of WTAP gene and the clinical characteristics was analyzed. The relative mRNA expression of WTAP gene in the non-M3 AML group was significantly higher than that in the healthy control group, and the difference showed statistically significant (P<0.01). There showed no statistically significant difference in WTAP gene expression among each subtypes (all P>0.05) according to the classification of FAB. The mRNA expression level of WTAP gene in FLT3-ITD mutated AML patients was higher than that in FLT3-ITD unmutated group (P=0.016), and the mRNA expression level of WTAP gene in AML patients with CEBPα mutation was lower than that in CEBPα unmutated group (P=0.016). The expression level of WTAP mRNA was positively correlated with WT1 expression (r=0.6866, P<0.01). There was no relationship between WTAP mRNA expression level and other clinical parameters, such as age, gender, white blood cell count, hemoglobin level, platelet count, bone marrow original proportion of immature cells, chromosome karyotype, and NPM1, DNMT3A, ASXL1, NRAS, TET2 genes mutation status (P>0.05). The expression level of WTAP mRNA showed no obvious effect on the complete remission of patients after first treatment. The different expression level of WTAP gene at initial diagnosis showed also no effect on the overall survival time of patients. The expression level of WTAP gene is increasing in new diagnosed non-M3 acute myeloid leukemia. There is a positive correlation between the expression level of WTAP gene and the expression level of WT1 fusion gene. WTAP mRNA always shows higher expression in patients with FLT3-ITD mutation than that in patients without FLT3-ITD mutation, and shows lower expression in patients with CEBPα mutation than that in unmutated group.