Prognostic Factor In Acute Myeloid Leukemia Patients Research Articles

CSRP1 gene: a potential novel prognostic marker in acute myeloid leukemia with implications for immune response

BackgroundAcute myeloid leukemia, constituting a majority of leukemias, grapples with a 24% 5-year survival rate. Recent strides in research have unveiled fresh targets for drug therapies. LIM-only, a pivotal transcription factor within LIM proteins, oversees cell development and is implicated in tumor formation. Among these critical LIM proteins, CSRP1, a Cysteine-rich protein, emerges as a significant player in various diseases. Despite its recognition as a potential prognostic factor and therapeutic target in various cancers, the specific link between CSRP1 and acute myeloid leukemia remains unexplored. Our previous work, identifying CSRP1 in a prognostic model for AML patients, instigates a dedicated exploration into the nuanced role of CSRP1 in acute myeloid leukemia.MethodsR tool was conducted to analyze the public data. qPCR was applied to evaluate the expression of CSRP1 mRNA for clinical samples and cell line. Unpaired t test, Wilcoxon Rank Sum test, KM curves, spearman correlation test and Pearson correlation test were included in this study.ResultsCSRP1 displays notable expression variations between normal and tumor samples in acute myeloid leukemia (AML). It stands out as an independent prognostic factor for AML patients, showing correlations with clinical factors like age and cytogenetics risk. Additionally, CSRP1 correlates with immune-related pathways, immune cells, and immune checkpoints in AML. Furthermore, the alteration of CSRP1 mRNA levels is observed upon treatment with a DNMT1 inhibitor for THP1 cells.ConclusionThe CSRP1 has potential as a novel prognostic factor and appears to influence the immune response in acute myeloid leukemia. Additionally, there is an observed association between CSRP1 and DNA methylation in acute myeloid leukemia.

Metastasis Related Epithelial-Mesenchymal Transition Signature Predicts Prognosis and Response to Chemotherapy in Acute Myeloid Leukemia.

Acute myeloid leukemia (AML) is a highly heterogenous disease with varying clinical outcomes among patients. Epithelial-mesenchymal transition (EMT) is an important mechanism underlying cancer metastasis and chemotherapy resistance. However, few EMT-based signatures have been established to predict AML prognosis and treatment efficacy. By conducting comparative RNA-seq analysis, we discovered the differential expression of EMT genes between AML patients with relapse and those without relapse. Based on the prognostic analysis of the differentially expressed EMT genes, a metastasis-related EMT signature (MEMTs) was constructed. An analysis was conducted on both TARGET and TCGA cohorts to explore the possible association between MEMTs and prognosis in AML. Three separate chemotherapy treatment cohorts were utilized to assess the predictive efficacy of MEMTs for chemotherapy response. In addition, the potential correlation between MEMTs and the tumor microenvironment was also investigated. Finally, random forest analysis and functional experiments were conducted to verify the key MEMTs gene associated with AML metastasis. Based on expression and prognostic analysis, we constructed MEMTs that include three EMT genes (CDH2, LOX, and COL3A1). Our findings suggested that the MEMTs could act as a prognostic factor for AML patients, and furthermore, it proved to be a predictor of their response to chemotherapy. Specifically, high MEMTs was associated with worse prognosis and poor response to chemotherapy, while low MEMTs was linked to better prognosis and higher response rates. Random forest and functional experiments demonstrate that CDH2 is a key gene promoting leukemia cell metastasis among the three MEMTs genes. The identification of MEMTs could potentially act as a predictor for the prognosis and the response to chemotherapy in AML patients. Individual tumor evaluation based on MEMTs could provide personalized treatment options for AML patients in the future.

The prognostic value of RASGEF1A RNA expression and DNA methylation in cytogenetically normal acute myeloid leukemia.

Acute myeloid leukemia (AML) is a significantly heterogeneous malignancy of the blood. Cytogenetic abnormalities are crucial for the prognosis of AML. However, since more than half of patients with AML are cytogenetically normal AML (CN-AML), predictive prognostic indicators need to be further refined. In recent years, gene abnormalities are considered to be strong prognostic factors of CN-AML, already having clinical significance for treatment. In addition, the relationship of methylation in some genes and AML prognosis predicting has been discovered. RASGEF1A is a guanine nucleotide exchange factors of Ras and widely expressed in brain tissue, bone marrow and 17 other tissues. RASGEF1A has been reported to be associated with a variety of malignant tumors, examples include Hirschsprung disease, renal cell carcinoma, breast cancer, diffuse large B cell lymphoma, intrahepatic cholangiocarcinoma and so on [1, 2]. However, the relationship between the RASGEF1A gene and CN-AML has not been reported. By integrating the Cancer Genome Atlas (TCGA) database 75 patients with CN-AML and 240 Gene Expression Omnibus (GEO) database CN-AML samples, we examined the association between RASGEF1A's RNA expression level and DNA methylation of and AML patients' prognosis. Then, we investigated the RASGEF1A RNA expression and DNA methylation's prognostic value in 77 patients with AML after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) as well as 101 AML patients after chemotherapy respectively. We investigated the association between sensitivity to Crenolanib and expression level of RASGED1A in patients by integrating 191 CN-AML patients from BeatAML dadataset. We integrated the expression and methylation of RASGEF1A to predict the CN-AML patients' prognosis and investigated the relationship between prognostic of AML patients with different risk classification and expression levels or methylation levels of RASGEF1A. We found that RASGEF1A gene high expression group predicted poorer event-free survival (EFS) (P< 0.0001) as well as overall survival (OS) (P< 0.0001) in CN-AML samples, and the identical results were found in AML patients receiving chemotherapy (P< 0.0001) and Allo-HSCT (P< 0.0001). RASGEF1A RNA expression level is an CN-AML patients' independent prognostic factor (EFS: HR = 5.5534, 95% CI: 1.2982-23.756, P= 0.0208; OS: HR = 5.3615, 95% CI: 1.1014-26.099, P= 0.0376). The IC50 (half maximal inhibitory concentration) of Crenolanib of CN-AML samples with RASGEF1A high expression level is lower. In addition, patients with high RASGEF1A methylation level had significant favorable prognosis (EPS: P< 0.0001, OS: P< 0.0001). Furthermore, the integrative analysis of expression and methylation of RASGEF1A could classify CN-AML patients into subgroups with different prognosis (EFS: P= 0.034, OS: P= 0.0024). Expression levels or methylation levels of RASGEF1A help to improve risk classification of 2010 European Leukemia Net. Higher RASGEF1A RNA expression and lower DNA methylation predicts CN-AML patients' poorer prognosis. The RASGEF1A high expression level from patients with CN-AML have better sensitivity to Crenolanib. The integrative analysis of RASGEF1A RNA expression and DNA methylation can provide a more accurate classification for prognosis. Lower RASGEF1A expression is a favorable prognostic factor for AML patients receiving chemotherapy or Allo-HSCT. 2010 European Leukemia Net's risk classification can be improved by RASGEF1A expression levels or methylation levels.

The Expression of mi-RNAs Involved in the Hematopoietic Niche Microenvironment in Newly Diagnosed AML Patients

Introduction Acute myeloid leukemia (AML) is a genetically heterogeneous disease and the most common type of acute leukemia in adults. Despite the progress made in the last years in AML therapy, induction treatment failure and disease relapse still remain the main challenges in clinical practice. miRNAs are involved in the pathogenesis of AML as well as influence the differentiation and proliferation of normal hematopoietic cells. The expressions of specific miRNAs have been identified as potential biomarkers for predicting prognosis both in solid tumors and hematological malignancies. Methods The expression of 12 miRNAs involved in hematopoiesis and the microenvironment of the hematopoietic niche: miR-15a-5p, miR-34a-5p, miR-125b-5p, miR-139-5p, miR-146-5p, miR-181a-5p, miR-199b-5p, miR-204-5p, miR-218-5p, miR-299-5p, miR-424-3p, miR-452-5p was determined in duplicates using qPCR in bone marrow samples of newly diagnosed AML patients. The miRCURY LNA SYBR® Green PCR Kit (Qiagen) was used to carry out qPCR. miRNAs expression was normalized using the mean expression of all miRNAs in a given sample. The normalization was performed using the formula ∆Ct = Ct(reference) − Ct(miRNA of interest). This approach results in higher values for higher miRNA expression. Differential expression analysis was carried out using a t-test. Cox proportional hazard regression analyses of clinical factors and miRNAs for predicting overall survival in AML patients were performed. Results To the study group we enrolled 45 (23 women and 22 men) newly diagnosed AML adults with the mean age 45.1 ± 12.8 years. Among them, 13.9% patients (pts) had a low-risk, 44.2% intermediate-risk and 41.9% high-risk AML according to EuropeanLeukemiaNet (ELN) 2017 risk stratification (Döhner et al. 2017). In the study group, 16.7% of pts had NPM1 mutation, whereas 30.2% of pts had FLT3-ITD or FLT3-TKD mutation. The majority of patients (88.9%) received intensive treatment: DA-60 (daunorubicin 60mg/m2 + cytarabine 200mg/m2) regimen (50%), followed by DAC (DA-60 + cladribine 5mg/m2) (40%) and DA-90 (daunorubicin 90mg/m2 + cytarabine 100mg/m2) (10%). Azacitidine in standard dose was used as the low-intensity regimen. There were no statistically significant differences in miRNAs expression according to complete remission (CR) achievement. Patients with high-risk AML had higher expression of miR-125b-5p (fold change- FC=2.3, p=0.017) and lower expression of miR-34a-5p (FC=0.3, p=0.029), miR-199b-5p (FC=0.5, p=0.039) and miR-15a-5p (FC=0.6, p=0.043) as compared to low- and intermediate-risk pts. In patients with NPM1 mutation, miR-34a-5p, miR-146a-5p and miR-15a-5p were significantly upregulated, whereas miR-139-5p, miR-424-3p and miR-452-5p were downregulated. FLT-3-mutated patients had significantly lower expression of miR-424-3p (FC=0.2, p=0.019) comparing to FLT-3-unmutated controls. In patients with unfavorable karyotype, upregulation of miR-125b-5p (FC=2.8, p=0.002) and miR-139-5p (FC=3.0, p=0.027) was observed meanwhile lower expression of miR-34a-5p (FC=0.3, p=0.035) in these patients was detected. The median overall survival (OS) in the study cohort was 12.3 months. In univariate Cox regression analysis, hematopoietic cell transplantation-comorbidity index (HCTCI) score >0 (HR 2.86, 95%CI: 1.15-7.12, p=0.024), white blood cell (WBC) count at diagnosis (HR 1.01, 95%CI: 1.00-1.01, p=0.033), and higher expression (>median) of miR-199b-5p (HR 0.31, 95%CI: 0.12-0.81, p=0.027) (Fig.1) were factors influencing the outcome. In multivariate model, higher expression of miR-199-5p (HR 0.27, 95%CI: 0.08-0.91, p=0.036) retained their significant protective effect in the context of established prognostic factors. Conclusions Our preliminary results demonstrate that the expression of miRNAs involved in the microenvironment niche is deregulated in AML patients. The upregulation of miR-199-5p at the diagnosis is associated with longer overall survival of AML patients. Significantly lower expression of miR-199-5p was reported in ELN high-risk group. These data indicate that miR-199-5p may be a potential prognostic factor in AML patients. Larger studies are needed to confirm our observation. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Pan-cancer analysis of oncogenic TNFAIP2 identifying its prognostic value and immunological function in acute myeloid leukemia

BackgroundTumor necrosis factor alpha-induced protein 2 (TNFAIP2), a TNFα-inducible gene, appears to participate in inflammation, immune response, hematopoiesis, and carcinogenesis. However, the potential role of TNFAIP2 in the development of acute myeloid leukemia (AML) remains unknow yet. Therefore, we aimed to study the biological role of TNFAIP2 in leukemogenesis.MethodsTNFAIP2 mRNA level, prognostic value, co-expressed genes, differentially expressed genes, DNA methylation, and functional enrichment analysis in AML patients were explored via multiple public databases, including UALCAN, GTEx portal, Timer 2.0, LinkedOmics, SMART, MethSurv, Metascape, GSEA and String databases. Data from The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO) and Beat AML database were used to determine the associations between TNFAIP2 expression and various clinical or genetic parameters of AML patients. Moreover, the biological functions of TNFAIP2 in AML were investigated through in vitro experiments.ResultsBy large-scale data mining, our study indicated that TNFAIP2 was differentially expressed across different normal and tumor tissues. TNFAIP2 expression was significantly increased in AML, particularly in French–American–British (FAB) classification M4/M5 patients, compared with corresponding control tissues. Overexpression of TNFAIP2 was an independent poor prognostic factor of overall survival (OS) and was associated with unfavorable cytogenetic risk and gene mutations in AML patients. DNA hypermethylation of TNFAIP2 at gene body linked to upregulation of TNFAIP2 and inferior OS in AML. Functional enrichment analysis indicated immunomodulation function and inflammation response of TNFAIP2 in leukemogenesis. Finally, the suppression of TNFAIP resulted in inhibition of proliferation by altering cell-cycle progression and increase of cell death by promoting early and late apoptosis in THP-1 and U937AML cells.ConclusionCollectively, the oncogenic TNFAIP2 can function as a novel biomarker and prognostic factor in AML patients. The immunoregulation function of TNFAIP2 warrants further validation in AML.

IER3 (IEX-1) dysregulation serves as a potential prognostic factor in acute myeloid leukemia patients.

IntroductionImmediate early response 3 (IER3) has association with hematological malignancies’ risk and prognosis, such as myelodysplastic syndrome, while its relation to acute myeloid leukemia (AML) is not clear. This study aimed to explore the correlation of IER3 with AML risk, clinical characteristics, complete remission (CR), event‐free survival (EFS), and overall survival (OS).MethodsA total of 93 de novo AML patients were included in this study. In addition, 30 patients with non‐hyperplasia hematologic malignancies requiring bone marrow testing (as disease controls) and 30 health donors (as health controls) were also recruited. Bone morrow samples of AML patients (before treatment), disease controls (before treatment), and health controls (at donation) were collected. IER3 in bone marrow mononuclear cells was detected by reverse transcription‐quantitative polymerase chain reaction.ResultsIER3 was increased in AML patients compared with disease controls and health donors (both P < .001), and receiver operating characteristic (ROC) curve showed that IER3 had certain capability of distinguishing AML patients from disease controls (area under curve (AUC): 0.735, 95% confidence interval (CI): 0.650‐0.820), and health donors (AUC: 0.789, 95% CI: 0.712‐0.866). Meanwhile, IER3 was correlated with FLT3‐ITD mutation (P = .030) and poor NCCN risk stratification (P = .031) in AML patients. Moreover, IER3 had negative association with CR in AML patients (P = .022), and showed certain potential in discriminating CR patients from non‐CR patients (AUC: 0.655, 95% CI: 0.533‐0.777). Besides, IER3 was negatively associated with EFS (P = .033), but not OS (P = .083) in AML patients.ConclusionIER3 dysregulation serves as a potential prognostic factor in AML patients.

Effect of long non-coding RNA MBNL1-AS1 expression on prognosis of acute myeloid leukemia patients

Objective: To explore the prognosis effect of the expression of long-chain non-coding RNA (lncRNA) MBNL1-AS1 on acute myeloid leukemia (AML) patients. Methods: One hundred and twenty-five AML patients of the Cancer Genome Atlas (TCGA) from November 2001 to March 2010 were involved, including 70 patients who received chemotherapy only and other 55 patients treated with allogeneic hematopoietic stem cell transplantation (allo-HSCT) in addition to chemotherapy. According to the median expression of lncRNA MBNL1-AS1, patients of chemotherapy group were divided into high expression sub-group(n=35) and low expression sub-group (n=35), and patients of allo-HSCT group were also divided into high expression sub-group (n=28) and low expression sub-group (n=27) for prognosis analysis. Clinical characteristics at diagnosis, including peripheral white blood cell counts (WBC), blast percentages in peripheral blood and bone marrow (BM), French-American-British (FAB) subtypes and the frequencies of common genetic mutations in AML were described. The event-free survival (EFS) rate and overall survival (OS) rate of patients in different groups were analyzed, and the influence of the clinical characteristics of patients on the prognosis of AML was analyzed by COX multivariate analysis. Results: In the chemotherapy group, patients with low lncRNA-MBNL1-AS1 expression had significantly lower EFS and OS (60.0%, 8.6%) than patients with high lncRNA-MBNL1-AS1 expression (68.6%, 34.3%) (χ²=7.817, 10.880, all P<0.01). However, in the alloHSCT group, no significant differences were observed in EFS and OS of patients between high and low expression groups of lncRNA-MBNL1-AS1 (all P>0.05). COX multivariate analysis confirmed that age≥60 years old (EFS: HR (95%CI): 6.934 (1.918-25.075),P=0.003;OS: HR (95%CI): 4.119 (1.812-9.364), P=0.001), and low expression of lncRNA MBNL1-AS1 (EFS: HR (95%CI): 0.354 (0.126-0.941), P=0.038; OS: HR (95%CI): 0.424 (0.231-0.778), P=0.006)were independent risk factors for EFS and OS in the chemotherapy group. Conclusion: The long-chain non-coding RNA MBNL1-AS1 is related to the prognosis of AML, and its low expression is an independent poor prognostic factor in AML patients.

Prognostic Value of Interleukin-17 A Gene Polymorphism and Serum IL-17 Levels in Adult Acute Myeloid Leukaemia Patients

Background: T helper 17 (Th17) cells play a role in the pathogenesis and prognosis of acute myeloid leukemia (AML). Th17 cells produce interleukin (IL)-17A and IL-17F, that share strong surface expression and homology of the IL-23 receptor (IL-23R). Objective: To evaluate the expression of IL-17A gene polymorphism and IL-17 serum level in adult AML patients and to clarify its prognostic significance. Patients and methods: In this prospective study, 48 patients with de novo AML and 48 healthy matched controls were enrolled. Cases were diagnosed at clinical pathology and managed in Medical Oncology Departments. We evaluated both serum IL-17 levels, measured by ELISA, and IL-17A (rs2275913) gene polymorphism was assessed using realtime polymerase chain reaction (PCR) in all patients and control. All patients received “3+7” as induction, responders received 3-4 courses of high dose cytarabine as consolidation, while non-responders received salvage therapy. Results: Serum levels of IL-17 were higher in AML patients (median 27 pg/ml) vs 17 pg/ml in the control group (P < 0.001). Also, serum IL 17 level was higher in non-responders with median 37.5 (pg/ml) vs 23 (pg/ml) in responders (P < 0.001). while IL-17A mutant genotypes and alleles showed no significant relation between expression of IL-17 A gene polymorphism and response to treatment or outcome of studied AML cases. Conclusion: Serum IL-17 levels can be considered a useful diagnostic and prognostic factor in AML patients, unlike IL-17 A gene polymorphisms.

Prognostic Value of Interleukin-17 A Gene Polymorphism and Serum IL-17 Levels in Adult Acute Myeloid Leukaemia Patients

Background: T helper 17 (Th17) cells play a role in the pathogenesis and prognosis of acute myeloid leukemia (AML). Th17 cells produce interleukin (IL)-17A and IL-17F, that share strong surface expression and homology of the IL-23 receptor (IL-23R). Objective: To evaluate the expression of IL-17A gene polymorphism and IL-17 serum level in adult AML patients and to clarify its prognostic significance. Patients and methods: In this prospective study, 48 patients with de novo AML and 48 healthy matched controls were enrolled. Cases were diagnosed at clinical pathology and managed in Medical Oncology Departments. We evaluated both serum IL-17 levels, measured by ELISA, and IL-17A (rs2275913) gene polymorphism was assessed using realtime polymerase chain reaction (PCR) in all patients and control. All patients received “3+7” as induction, responders received 3-4 courses of high dose cytarabine as consolidation, while non-responders received salvage therapy. Results: Serum levels of IL-17 were higher in AML patients (median 27 pg/ml) vs 17 pg/ml in the control group (P < 0.001). Also, serum IL 17 level was higher in non-responders with median 37.5 (pg/ml) vs 23 (pg/ml) in responders (P < 0.001). while IL-17A mutant genotypes and alleles showed no significant relation between expression of IL-17 A gene polymorphism and response to treatment or outcome of studied AML cases. Conclusion: Serum IL-17 levels can be considered a useful diagnostic and prognostic factor in AML patients, unlike IL-17 A gene polymorphisms.

Role of CD25 expression on prognosis of acute myeloid leukemia: A literature review and meta-analysis.

The gene expression for interleukin-2 receptor subunit alpha (CD25/IL2RA) is frequently altered in adults with acute myeloid leukemia (AML). Increasing evidence indicates that the elevated expression of CD25 may be correlated with poor survival for AML patients. Thus, we performed this meta-analysis to further evaluate the prognostic value of elevated CD25 in AML. Eligible studies were gathered by searching on PubMed, Web of Science, and Embase. Using the R language 3.6.0 software, Pooled hazard ratios (HRs) with their corresponding 95% confidence intervals (CIs) of overall survival (OS) and disease-free survival (DFS)/relapse-free survival (RFS)/event-free survival (EFS) for total and subgroup analyses were calculated to investigate the association of elevated CD25 and outcomes of AML patients. Ten studies with a total of 1640 participants were enrolled in this meta-analysis. Pooled HRs suggested that overexpression of CD25 predicted poor outcomes on both OS (HR = 2.27, 95%CI 1.95–2.64) and DFS/RFS/EFS (HR = 1.77, 95%CI 1.44–2.17) in overall population. Subgroup analyses stratified by ethnicity, AML subtype, cut-off value, statistical methodologies and detection method draw similar results. Our meta-analysis indicates that elevated CD25 expression is a poor prognostic factor for AML patients. Considering limited number of samples, further relevant studies are warranted.

HLA Evolutionary Divergence as a Prognostic Marker for AML Patients Undergoing Allogeneic Stem Cell Transplantation.

The diversity of human leukocyte antigens (HLAs) enables the presentation of immense repertoires of peptides, including tumor-associated antigens (TAAs). As a surrogate for immunopeptidome diversity, the HLA evolutionary divergence (HED) between individual HLA alleles might directly define the ability to present TAAs, a prerequisite for graft-versus-leukemia effects. We therefore analyzed the impact of HED on survival within a cohort of 171 acute myeloid leukemia (AML) patients after matched donor allogeneic hematopoietic stem cell transplantation (HSCT). Low HED (<25th percentile) of HLA class I (HEDclass I) or HLA-DR antigens (HEDDR) was a strong determinant for adverse overall survival after allogeneic HSCT (OS), with a hazard ratio for death of 1.9 (95% CI 1.2–3.2) and 2.1 (95% CI 1.3–3.4), respectively. Defining a cutoff value for the combined HEDtotal (HEDclass I and HEDDR), the respective 5 year OS was 29.7% and 64.9% in patients with low and high HEDtotal (p < 0.001), respectively. Furthermore, the risk of relapse was significantly higher in patients with low HEDtotal (hazard ratio (HR) 2.2, 95% CI 1.3–3.6) and event-free survival (EFS) was significantly reduced (5 year EFS 25.7% versus 54.4%, p < 0.001). We here introduce HED, a fundamental metric of immunopeptidome diversity, as a novel prognostic factor for AML patients undergoing allogeneic HSCT.

FAM213A is linked to prognostic significance in acute myeloid leukemia through regulation of oxidative stress and myelopoiesis.

Accurate prediction of malignancies is important in choosing therapeutic strategies. Although there are many genetic and cytogenetic prognostic factors for acute myeloid leukemia (AML), prognosis is difficult to predict because of the heterogeneity of AML. Prognostic factors, including messenger RNA (mRNA) expression, have been determined for other malignancies, but not for AML. A total of 402 patients from The Cancer Genome Atlas, GSE12417 (GPL96, 97), and GSE12417 (GPL570) were included in this study. In Kaplan-Meier curve analyses, high expression of family with sequence similarity 213 member A (FAM213A), which activates antioxidant proteins, was associated with worse prognosis of AML. Similar to the results of the survival curve, C-indices and area under the curve values were high. Current prognostic factors of AML, unlike those of other cancers, do not take mRNA expression into consideration. Thus, the development of mRNA-based prognostic factors would be beneficial for accurate prediction of the survival of AML patients. Additionally, in vivo validation using zebrafish revealed that fam213a is important for myelopoiesis at the developmental stage and is a negative regulator of the p53 tumor suppressor gene. The findings implicate fam213a as a novel prognostic factor for AML patients.

Expression of WT1 gene and its prognostic value in patients with acute myeloid leukemia

To investigate the expression of Wilms'tumor 1 (WT1) gene in patients with acute myeloid leukemia (AML), and to explore its application in predicting prognosis of AML in patients with wild or mutated nucleophosmin 1(NPM1) and Fms-like tyrosine kinase 3-internal tandem duplication (FLT3-ITD). One hundred and sixty-seven newly diagnosed AML patients(exclued M3 type) were enrolled in this study. The survival of patients were analyzed with Kaplan-Meier method. The clinical data, laboratory findings and the survival of patients were analyzed and compared between patients with high WT1 expression (high-WT1 group) and those with low WT1 expression (low-WT1 group), as well as among the patients with NPM1 or FLT3-ITD wild type and mutants. The overall response rates (ORR) in high-WT1 and low-WT1 groups were 65.9% (83/126) and 95.1% (39/41), respectively (P<0.01). Compared with the low-WT1 group, the high-WT1 group had lower 2-y overall survival (OS) rate (46.1% vs. 75.2%, P<0.05) and 2-y disease free survival (DFS) rate (43.5% vs. 68.5%, P<0.05). After induction chemotherapy, the patients with decreased WT1 gene expression ≥ 1log was associated with higher ORR and 2-y OS rate (all P<0.05), but the advantage of 2-y DFS rate was not shown (P>0.05). In patients with NPM1 wild type, the high-WT1 group had inferior ORR and 2-y OS rate (all P<0.05), while in the patients with FLT3-ITD wild type, the high-WT1 group had inferior ORR, 2-y OS rate and 2-y DFS rate (all P<0.05). In patients with NPM1 or FLT3 -ITD mutations, the WT1 expression had no significantly predicting values in treatment efficacy and survival (all P>0.05). WT1 gene overexpression indicated poor prognosis of AML patients; the patients with decreased WT1 gene expression ≥ 1 log after the first induction therapy show better prognosis than those with<1 log. The WT1 gene expression level at diagnosis can be used as an unfavorable prognostic factor for AML patients with NPM1 or FLT3-ITD wild types.

MiR-592 functions as a tumor suppressor in acute myeloid leukemia by targeting ROCK1 and predicts patients' prognosis.

Dysregulation of miR-592 has been reported in several tumors. However, its role in acute myeloid leukemia (AML) remains unknown. The present study aimed at investigating the expression pattern and biological function of miR-592 in AML and to elucidate the mechanism involved. qRT-PCR was used to analyze the expression of miR-592 in bone marrow and serum obtained from AML patients and healthy controls. The associations between serum miR-592 expression and clinical features and prognosis of AML patients were statistically analyzed. Then we detected the effect of miR-592 on proliferation, metastasis and apoptosis by CCK-8 assay, Transwell assays and flow cytometry, respectively. Dual-luciferase reporter assays were performed to validate the regulation of a putative target of miR-592. Rescue experiments were performed to confirm whether ROCK1 was a direct and functional target of miR-592 in AML. We found that the expression level of miR-592 was significantly lower in AML patients and AML cell lines. Low expression of serum miR-592 was associated with advanced French-American-British classification, cytogenetics and poor prognosis. Multivariate analysis confirmed that serum miR-592 expression was an independent prognostic factor for AML patients. Functionally, overexpression of miR-592 suppressed AML cell growth and metastasis, and promoted apoptosis. Further mechanistic investigation showed ROCK1 was a direct target gene of miR-592. Finally, ROCK1 overexpression rescued the effect of miR-592-mediated AML cell proliferation and metastasis. These findings suggest that miR-592 acted as a tumor suppressor by targeting ROCK1 and may serve as a potential biomarker in AML.

FLT3-ITD compared with DNMT3A R882 mutation is a more powerful independent inferior prognostic factor in adult AML patients after Allogeneic Hematopoietic Stem Cell Transplantation: A retrospective cohort study

ObjectiveThis study aimed to evaluate DNMT3A exon 23 mutations and their prognostic impacts in the presence of NPM1 and FLT3 mutations in acute myeloid leukemia (AML) patients who underwent allogeneic hematopoietic stem cell transplantation (HSCT).Materials and MethodsThis study comprised 128 adult AML patients referred to the Hematology-Oncology and Stem Cell Research Center of Shariati Hospital. NPM1 and FLT3-ITD mutations were detected by fragment analysis. For DNMT3A exon 23 mutation analysis, we used Sanger sequencing. Overall survival (OS) and relapse-free survival (RFS) curves were estimated by the Kaplan-Meier method and the log-rank test was used to calculate differences between groups.ResultsThe prevalence of DNMT3A exon 23 mutations was 15.6% and hotspot region R882 mutations were prominent. RFS and OS were compared in patients with and without DNMT3A exon 23 mutations using univariate analysis and there was no significant difference between these groups of patients. On the contrary, the FLT3-ITD mutation significantly reduced the OS (p=0.009) and RFS (p=0.006) in AML patients after allogeneic HSCT. In the next step, patients with AML were divided into four groups regarding FLT3-ITD and DNMT3A mutations. Patients with DNMT3A R882mut/FLT3-ITDpos had the worst OS and RFS. These results indicate that DNMT3A mutations alone do not affect the clinical outcomes of AML patients undergoing allogeneic HSCT, but when accompanied by FLT3-ITD mutations, the OS was significantly reduced (5-year OS 0% for DNMT3A R882mut/FLT3-ITDpos patients vs. 62% DNMT3A R882wt/FLT3-ITDneg, p=0.025) and the relapse rate increased.ConclusionIt can be deduced that DNMT3A R882mut/FLT3-ITDpos is an unfavorable prognostic factor in AML patients even after allogeneic HSCT.

Reduced expression of chemerin is associated with poor clinical outcome in acute myeloid leukemia.

Chemerin is dysregulation in numerous solid cancers. However, only little is known about the role of chemerin in acute myeloid leukemia (AML). In this study, we aimed to investigate the expression and clinical significance of recently described chemerin in acute myeloid leukemia (AML). The expression of chemerin in 149 patients with de novo AML and 35 normal controls was quantified by Real-time quantitative PCR (RQ-PCR). Chemerin was down-expressed in AML compared with controls (P=0.042). A receiver operating characteristic (ROC) curve revealed that chemerin expression could differentiate patients with AML from control subjects (AUC=0.611, 95% CI: 0.490-0.732; P=0.042) respectively. The cohort of AML patients was divided into two groups according to the cut-off value of 0.0826 (79% sensitivity and 54% specificity, respectively). In addition, the AML patients with low chemerin expression had significantly shorter overall survival (OS) than those with high chemerin expression (P=0.049). Moreover, multivariate survival analysis confirmed that chemerin was an independent prognostic factor for AML patients. In conclusion, downregulation of chemerin might be a useful diagnostic and prognostic factor for AML patients.

High Expression of ARMCX1 Predicts Poor Survival in Intensively Treated Older Acute Myeloid Leukemia Patients (≥ 60 years)

The Armadillo repeat-containing X-linked protein 1 (ARMCX1)gene encodes a member of the ALEX family of proteins and is located on the X chromosome. We found this gene aberrantly expressed in acute myeloid leukemia (AML) cells.We analyzed ARMCX1 expression in a cohort of 508 AML patients who received intensive chemotherapy within AMLCG clinical trial. Expression levels of ARMCX1 were determined in pretreatment bone marrow or peripheral blood samples by using oligonucleotide microarrays (GSE37642). High transcript levels (dichotomized at the median expression) of ARMCX1 were predictors for inferior overall survival (OS) (p=0.0027, Log-Rank test) and a lower rate of complete remission (CR) (p=0.0017). Older patients (≥ 60 years) showed an elevated incidence of high ARMCX1 expression compared to younger patients (<60 years) (56% vs. 44%, p=0.008, Chi-Squared test). In the subgroup analyses we found that for younger AML patients (<60 years) ARMCX1 expression was not predictive for survival (OS, PFS, EFS) and CR. However, in older patients (≥ 60 years) OS (p=0.002, median survival 11 vs 8 months), event free survival (EFS) (p=0.0006, median 6 vs 2 months) and progression free survival (PFS) (p=0.013, median 10 vs 7 months) were significantly lower among patients with high ARMCX1 expression. CR rate of elderly AML patients with low ARMCX1 expression levels was 60% vs. 41% with high levels (P=0.003). Older AML patients with low ARMCX1 expression had a 5-year OS of 20% (vs. 7% for patients with high expression). Furthermore, prognostic significance of high ARMCX1 expression was confirmed in an independent AML patient cohort of 293 AML patients from the HOVON group (GSE6891). Both, OS (p<0.001) and EFS (p<0.001) were significantly inferior for patients with high ARMCX1expression in this cohort.Multivariate logistic regression did not show any association between ARMCX1 expression and sex, lactatdehydrogenase (LDH) or leukocyte count (WBC). Interestingly, high expression of ARMCX1 was strongly associated with high United Kingdom Medical Research Council (MRC) risk classification (p<0.0001).Evaluation of ARMCX1 expression against a clinical multivariate Cox-Regression model (age, gender) showed that dichotomized ARMCX1 added a significant amount of information to the model and was a risk factor for OS (p=0.0020, HR= 1.377 (95%-CI 1.124, 1.686)). Strikingly, including MRC risk classification into the multivariate Cox-Regression model high ARMCX1 expression remained a significant risk factor for OS (p=0.0413, HR=1.243 (95%-CI 1.009, 1.531)).Taken together, we analyzed ARMCX1 expression levels in a large AML patient cohort. In this study, high ARMCX1 expression levels were identified as a negative prognostic factor for AML patients with respect to survival and remission. DisclosuresHiddemann:Janssen: Membership on an entity's Board of Directors or advisory committees, Research Funding; Roche: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Gilead: Research Funding. Stelljes:Pfizer: Consultancy.

Association of WT1 rs16754 polymorphism with clinical features and prognosis in patients with acute myeloid leukemia

目的探讨WT1基因rs16754多态性与急性髓系白血病(AML)患者临床特征及预后的关系。方法收集2010年1月至2013年12月山西医科大学第二临床医学院115例AML患者初诊时骨髓及177名健康体检者外周血样本，采用高分辨熔解曲线法(HRM)进行WT1基因rs16754多态性位点分型，并通过直接测序法验证，回顾性分析该位点单核苷酸多态性(SNP)与AML发病风险、患者临床特征、缓解及生存情况的相关性。结果WT1基因rs16754多态性与AML发病风险无关(P= 0.296)，GA/AA组与GG组在患者性别、年龄及临床各指标间及化疗缓解率上差异均无统计学意义(P值均>0.05)，GA/AA组骨髓原始细胞比例高于GG组(P=0.025)。随访99例患者生存情况，单因素分析发现GA/AA组总体生存率高于GG组(P=0.035)。多因素分析显示A等位基因是AML患者的独立预后因素，GA/AA型患者的死亡风险高于GG型患者(HR=1.681,95% CI 1.046~2.700，P=0.032)。结论WT1 rs16754多态性与AML预后有关，可作为AML患者预后判断的指标之一。

The prognostic implication of SRSF2 mutations in Chinese patients with acute myeloid leukemia.

Recently, somatic mutations in SRSF2 gene have been discovered in a proportion of hematologic malignancies including acute myeloid leukemia (AML). This study was aimed to investigate SRSF2 mutations in Chinese AML patients. High-resolution melting analysis (HRMA) was developed to screen SRSF2 mutations in 249 cases with AML, and then direct DNA sequencing was used to verify the results of HRMA. In this study, 3.6% (9/249) of Chinese AML patients were found with heterozygous SRSF2 mutations. Patients with SRSF2 mutations were older than those with wild-type SRSF2 (P = 0.014). No differences in the sex, blood parameters, French-American-British classification (FAB) subtypes, and karyotypes were observed between AML patients with and without SRSF2 mutations. Although the overall survival (OS) of SRSF2-mutated patients was inferior to those without mutations in both whole AML patients (median 4 vs. 11months, respectively; P = 0.006) and cytogenetically normal patients (median 2 vs. 12months, respectively; P = 0.008), multiple analysis disclosed that SRSF2 mutation was not an independent prognostic factor in AML patients. These results suggest that SRSF2 mutation occurs at a low frequency in aged AML patients and might not be associated with adverse prognosis in Chinese AML patients.

Examination of the FLT3 and NPM1 mutational status in patients with acute myeloid leukemia from southeastern Poland.

IntroductionAcute myeloid leukemia (AML) is a genetically heterogeneous disease at both the cytogenetic and molecular levels. In AML cells many chromosomal aberrations are observed, some of them being characteristic of a particular subtype of patients, and others being less significant. Besides chromosomal abnormalities, the leukemic cells can have a variety of mutations involving individual genes. The aim of this work was to investigate the frequencies of molecular alterations with the focus on FLT3-ITD and NPM1 mutations in AML patients of different age groups living in a southeastern region of Poland.Material and methodsThe study group comprised 50 consecutive AML patients. We analyzed bone marrow samples by conventional cytogenetics. Cytogenetic evaluation in selected cases was complemented by the FISH technique. The internal tandem mutation in the FLT3 gene was identified using polymerase chain reaction (PCR), and the NPM1 mutation was assessed by direct nucleotide sequencing.ResultsThe studies using classical cytogenetics showed chromosomal aberrations in 32 (64%) patients. In 18 cases no changes in the karyotype were found by conventional karyotyping. FLT3-ITD mutation was detected in 4 (8%) patients and mutation of NPM1 in 3 patients with AML (6%).ConclusionsThe incidence of both mutations in our study group was lower than described elsewhere. We have confirmed that FLT3-ITD occurred more commonly in older patients and it was associated with shorter overall survival. By contrast, mutation of exon 12 of the NPM1 gene seems to be a good prognostic factor in AML patients with normal karyotype.