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Rao, GuoLi, Zhuxuanzi Wang, and Jiayu Liang. "Reinforcement Learning for Pattern Recognition in Cross-Border Financial Transaction Anomalies: A Behavioral Economics Approach to AML." Applied and Computational Engineering 142, no. 1 (2025): 116–27. https://doi.org/10.54254/2755-2721/2025.kl22287.
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This paper presents a novel approach to anti-money laundering (AML) in cross-border financial transactions by integrating reinforcement learning (RL) with behavioral economics principles. The research addresses critical limitations in traditional AML systems by conceptualizing money laundering detection as a sequential decision-making problem where detection policies adapt to evolving criminal strategies. We develop a specialized methodology that incorporates multi-level data representations, behavioral feature extraction algorithms, and a composite reward function designed to balance detection accuracy with investigation efficiency. The framework leverages behavioral economics principles to distinguish between legitimate financial anomalies and suspicious patterns indicative of money laundering activities. Experimental evaluation across three datasets demonstrates that the proposed approach achieves a 27.4% improvement in money laundering detection rate while reducing false alerts by 18.6% compared to state-of-the-art methods. Behavioral pattern recognition components prove particularly effective for identifying sophisticated laundering schemes characterized by strategic transaction structuring and temporal spacing designed to evade traditional detection systems. Case studies of cross-border money laundering operations validate the approach's effectiveness in operational environments. The research contributes a unified theoretical framework that enhances AML capabilities while providing practical implementation guidance for financial institutions and regulatory bodies engaged in combating cross-border financial crime.
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Kramarzova, Karolina, Andre Willasch, Bernd Gruhn, et al. "Expression Pattern of WT1 Isoforms in Patients with Acute Myeloid Leukemia (AML), Myelodysplastic Syndrome (MDS) and Severe Aplastic Anemia (SAA)." Blood 118, no. 21 (2011): 2502. http://dx.doi.org/10.1182/blood.v118.21.2502.2502.
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Abstract Abstract 2502 Introduction: The Wilms' tumor gene 1 (WT1) is highly expressed in a large proportion of human acute leukemias and other hematological malignancies. It has been demonstrated that WT1 protein is produced in more than 36 different isoforms. These variants have distinct, partially overlapping functions and their ratio is supposed to influence the final effect of WT1. However, only limited information on WT1 isoforms has been published so far and the relevance of their expression pattern remains unclear. Aims: Our main aim was to determine the expression pattern of four WT1 isoforms characterized by the presence or absence of exon 5 and KTS insert (A[−/−], B[+/−], C[−/+], D[+/+]) in patients with AML, MDS and SAA. Materials and Methods: We designed a unique qPCR system for detection and quantification of 4 major WT1 isoforms. Using this method we analyzed the ratio of WT1 isoforms in 8 leukemic cell lines (Kasumi-1, NB-4, K562, MV4;11, REH, NALM6, UOCB6, RS4;11) and diagnostic BM samples of 73 childhood AML, 30 adult AML, 20 childhood MDS, 9 childhood SAA as well as 23 healthy controls. Results: Median expression level of total WT1 in healthy donors was 29 WT1/ABL×104 NCN. Childhood and adult AML expressed WT1 on significantly higher level compared to healthy controls (2058 and 3446 WT1/ABL×104 NCN respectively; p<0.0001). The expression level of total WT1 in patients with MDS was 196 WT1/ABL×104 NCN, whereas children with SAA had total WT1 comparable or lower than control samples (4 WT1/ABL×104 NCN). We found an excellent correlation between the total WT1 expression and the sum of WT1 isoforms (rho=0.916, p<0.0001). However, very low levels of total WT1 precluded detection of WT1 isoforms in majority of healthy donors and SAA samples, since we reached the limit of the sensitivity of qPCR method (as defined by limiting dilution experiments). For the same reason (very low total WT1 levels), 18 patients with AML (mostly AML M5) as well as 8 MDS samples were excluded from further analyses. The analysis of WT1 isoforms ratio showed a diverse expression patterns of WT1 variants in the particular cell lines (p<0.0001). Interestingly, a similar pattern of WT1 isoforms was present in cell lines with MLL/AF4 rearrangement, independent of the lineage (myeloid - MV4;11 and lymphoid - RS4;11). We could successfully determine the expression profile of WT1 isoforms in 1 healthy BM, which showed a substantial overexpression of isoform D with the ratio of 1.2: 1.8: 1.7: 5.3 for A, B, C and D variants, respectively. Rather surprisingly, we found a uniform expression pattern of WT1 isoforms (D>B>C∼A) in patients with childhood AML as well as adult AML (0.9: 2.5: 1.1: 5.5 and 1.3: 3.1: 1.3: 4.5 for isoforms A, B, C and D respectively). In contrast to AML samples, a different and more variable ratio of WT1 variants was found in children with MDS with predominance of variant D and similar levels of isoforms A, B and C (1.6: 1.5: 1.7: 4.6 for A, B, C and D respectively). Moreover, the unsupervised hierarchical cluster analysis according to the expression pattern of WT1 isoforms indicated that children with MDS belonged to a distinct cluster compared to childhood and adult AML. Conclusion: This is the first report of the analysis of WT1 isoforms expression pattern in AML and MDS using a unique qPCR method for the detection of WT1 variants. Our data suggest that WT1 isoforms expression pattern is surprisingly uniform in pediatric and adult AML with predominant expression of Exon5[+] isoforms. To the contrary, the ratio of WT1 isoforms in childhood MDS significantly differ from AML indicating that the expression pattern of WT1 variants is related to the type of malignant cells. Careful pre-analytical testing of the detection system parameters suggests the technical limitations in detection of WT1 isoforms in samples with very low total WT1 levels. Therefore, the previously reported differences in WT1 isoforms ratio between normal bone marrow and AML samples should be interpreted with caution. Supported by MSM0021620813, GAUK 81709, IGA NS10488–3/2009 Disclosures: No relevant conflicts of interest to declare.
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Reichle, Albrecht, Birgit Panzer, Gerlinde Goetz, et al. "Genetic Instability in Remission Hematopoiesis Is Different from That in AML Blasts: A New Diagnostic Tool to Study Biology of AML." Blood 104, no. 11 (2004): 1069. http://dx.doi.org/10.1182/blood.v104.11.1069.1069.
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Abstract Recently we have shown that aberrations, ie microsatellite instability (MSI) and allelic imbalance (AI) in positively selected CD34+ cells from leukapheresis products collected during first complete remission of de novo AML following double induction or consolidation according to AMLCG protocol may be predictive for relapse-free survival (RFS). Here we present data of an extended patient population treated on the same protocol. We compared MSI/AI pattern in CD34+ cells (n=60) with the corresponding pattern in AML blasts at diagnosis (n=60) or at relapse (n=12), and compared the AI/MSI pattern in CD34+ cells with those observed in the corresponding unselected bone marrow aspirates in first CR (n=50). The following loci were tested: D7S486, D7S525, D8S559, TP53, D11S1356, D2S123, APC, MfD5. Minimal residual disease (MRD) in the tested remission specimen (FACS analysis) was &lt;0.1%. MSI and/or AI at diagnosis could be detected in few cases (10 of 60, 17%), whereas CD34+ cells were tested positive in 25 of 60 cases (42%), and in 5 of 12 relapses studied (42%). Analysis of AI and MSI in DNA of bone marrow aspirates could not adequately confirm MSI and/or AI detectable in the corresponding CD34+ cells (n=3 cases, 12%). In 15 cases MSI and/or AI was observed in CD34+ cells, but not in AML blasts at diagnosis, in 5 cases in blasts at diagnosis, but not in CD34+ cells, in 4 cases in AML blasts at diagnosis and in CD34+ cells. Identical AI/MSI pattern were found in CD34+ cells and in AML blasts at relapse (n=2), as well as in AML blasts at diagnosis and relapse (n=1). According to the definition of genetic instability (GIN: MSI or &gt;2 AI at any locus, AI at APC locus), GIN in CD34+ cells was predictive for unfavorable RFS (p=0.001). In conclusion, detection of AI/MSI in CD34+ cells of remission hematopoiesis represents a new important diagnostic tool to study biology of AML besides the detection of MRD. For the first time we could demonstrate, that the same AIs or MSIs detectable in CD34+ cells, but not in AML blasts at diagnosis may occur in AML blasts at relapse.
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Chitra, P., M. R. Ebenezer Jebarani, P. Kavipriya, K. Srilatha, M. Sumathi, and S. Lakshmi. "Detection of AML in Blood Microscopic Images using Local Binary Pattern and Supervised Classifier." Research Journal of Pharmacy and Technology 12, no. 4 (2019): 1717. http://dx.doi.org/10.5958/0974-360x.2019.00286.5.
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Lingamfelter, Daniel, Na Yu, Tim Quinn, et al. "Detection and Production of Haptoglobin within Acute Myeloid Leukemic Blasts." Blood 110, no. 11 (2007): 4305. http://dx.doi.org/10.1182/blood.v110.11.4305.4305.
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Abstract While a directly proportional relationship between serum haptoglobin (Hp) levels and acute myeloid leukemia (AML) has been documented, the exact source of this elevated haptoglobin remains in dispute. Furthermore, no reported studies thus far have analyzed haptoglobin-targeted immunohistochemistry as a potential diagnostic tool for AML. In this study, we examined Hp expression in patients with AML. Previously collected blood samples were obtained, with informed consent, during routine diagnostic blood studies from twenty-one patients with AML (13 males, 8 females; age range 4–73 years). Cytospin samples were prepared, and immunohistochemistry was performed using a polyclonal anti-haptoglobin antibody. Total RNA was extracted from 19 AML patient samples as well as from a K562 leukemic cell line for the RT-PCR assay. Cytospin specimens derived from all twenty-one patients with confirmed AML showed consistent anti-haptoglobin staining of blast cells in a granular, cytoplasmic pattern. The K562 cell line, used as a positive control, stained in a similar fashion to the patient samples. Immunohistochemical co-labeling of blasts was achieved with the AML anti-podocalyxin marker in conjunction with the anti-haptoglobin antibody. All of the AML blasts as well as the K562 cells expressed Hp by the RT-PCR assay. The ratio index (RI) of Hp band density against β-actin serving as an internal control was used to compare the expression level among the AML cases. The RIs ranged from 0.89 to 0.02, indicating different Hp transcription levels among the AMLs. We conclude that haptoglobin is produced by AML blasts and stored within the cytoplasm. This may partly explain the increased serum level of haptoglobin observed in AML patients. Furthermore, immunohistochemistry consistently detects these intra-blastic haptoglobin stores, suggesting a potential for a novel diagnostic tool for AML.
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Haque, Salina, Zulfia Zinat Chowdhury, Tamanna Bahar, et al. "Immunophenotypic Characterisation Along with Aberrant Expression of CD markers in Morphologically Diagnosed Cases of Acute Myeloid Leukemia." Journal of Medicine 23, no. 2 (2022): 106–11. http://dx.doi.org/10.3329/jom.v23i2.60626.
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Immunophenotyping of leukemia cells is useful for detecting leukemia cell line, determining maturation stage and identifying aberrant antigens which act for individual treatment monitoring and detection of residual disease. A total of 104 newly diagnosed cases of acute myeloid leukemia were identified at hematology department in National Institute of Cancer Research and Hospital from January 2020 to December 2021.We detect Immunophenotypic pattern in newly diagnosed cases of acute myeloid leukemia. We also determine the frequency and pattern of aberrant expression of CD markers in acute myeloid leukemia patients. Mean age of patients was 35 years (SD±16 years) with male to female ratio was 1.53:1. Most frequent morphologic subtype was AMLM2 constituting 33.6% of all AML cases. Lineage specific markers HLADR, CD13, CD33, MPO, CD117 and CD34 were expressed in 80%, 89%, 95%, 77%, 74% and 62% cases of all AML cases respectively. Among 104 AML patient, aberrant CD expression was observed in 36% cases. The most frequently observed aberrant markers were CD7 and CD19 lymphoid markers, that were expressed in 15.38% and 14.42% cases respectively. Less frequent aberrant cCD3, CD10, CD5 and cCD79a antigens were expressed in 2.88%, 1.92%, 0.96% and 0.96% cases respectively. Immunophenotyping is essential in diagnosis and sub-classification of AML and expression of aberrant CD antigens is common in acute myeloid leukemia. These findings suggest the necessity for a more extensive study to evaluate the prognostic significance of aberrant CD marker expression in AML and to improve the accuracy of diagnosis and classification of AML. J MEDICINE 2022; 23: 106-111
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Wu, Kai, Qianyi Ma, Darren King, Jun Li, and Sami Malek. "Paired Analyses of AML at Diagnosis and Relapse By Single-Cell RNA Sequencing Identifies Two Distinct Relapse Patterns." Blood 134, Supplement_1 (2019): 183. http://dx.doi.org/10.1182/blood-2019-124170.
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Introduction: Despite achievement of complete remission (CR) following chemotherapy, Acute Myelogenous Leukemia (AML) relapses in the majority of adult patients. While relapsed AML is almost always clonally related to the disease at diagnosis, the actual molecular and cellular contributors to chemotherapy resistance and to AML relapse remain incompletely understood. Some molecular determinants of relapse have been identified in genomic, epigenetic and proteomic aberrations, while cellular relapse reservoirs have been identified in leukemia stem cells as well as in more mature leukemic cell compartments. Here, we set out to determine the cellular composition, gene mutation status and gene expression of paired AML specimens procured at diagnosis and at relapse aiming at a better understanding of the AML relapse process. Methods: We employed the drop-seq 3' single cell RNA sequencing (scRNA-seq) method (Macosko 2015) with minor modifications to analyze the mRNA expression in single cells derived from 12 paired AML specimens procured at diagnosis and at relapse from prior CR. We obtained scRNA-seq data on 1000-2000 single cells per sample detecting approximately 2000-3000 unique molecular identifiers (UMIs) and 800-1500 genes per cell. Using WES or panel-based sequencing we determined mutations in known driver genes. Subsequently, we optimized novel methods for detection and mapping of mutated driver genes to individual cells using mutation specific PCR conditions and novel bioinformatics approaches. We annotated scRNA-seq expression profiles of the diagnosis and relapsed AML specimens individually using publicly available data for cell type-specific RNA markers derived from sorted normal cell populations and further compared the scRNA-seq data to scRNA-seq data of 5 pooled normal human bone marrows generated for this study. Results: Through analyses of scRNA-seq data of paired diagnosis and relapse AML specimens via principle components analyses (PCA) or t-distributed stochastic neighbor embedding (t-SNE) we detected varying degrees of separation of cell clusters in all cases analyzed indicative of substantial changes in single cell gene expression between AML diagnosis and relapse. A few of these observed cluster shifts were paralleled by gain or loss of mutated genes (e.g. FLT3-ITD) at relapse while most others lacked obvious clonal genomic markers. Through subsequent comparison of the expression similarities of single AML cells to sorted normal human bone marrow cells we detected two distinct AML relapse patterns: i) a pattern of relapse suggesting simple leukemia regrowth as evidenced by similar proportions of leukemia cells mapping onto discrete normal bone marrow cells (e.g. monocyte-like or GMPs or CMPs), and, ii) a pattern of relapse whereby the gene expression of relapsed cells (but not diagnosis cells) had similarity to normal hematopoietic cells that are conventionally placed more apical in the classical hematopoiesis differentiation cascade (HSCs, MPPs, CMPs; a phenotypic shift to immaturity). In addition, no leukemia sample mapped to just one classically defined bone marrow cell type but instead to multiple cell types, suggesting that most AML leukemia cells harbor aberrant hybrid cell gene expression patterns. Finally, we detected quantitative shifts in T cells and NK cells in some samples at relapse, which will be analyzed in greater detail. Conclusions: The comparative analysis of scRNA-seq data of paired AML specimens procured at diagnosis and relapse, identifies frequent and previously unrecognized changes in gene expression in leukemia cells at relapse. Through a comparison of gene mutation and gene expression at single cell resolution we identify two distinct AML relapse patterns in adult AML. Disclosures No relevant conflicts of interest to declare.
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Orsmark-Pietras, Christina, Henrik Lilljebjörn, Marianne Rissler, et al. "Comprehensive Prospective Next Generation Sequencing of Acute Myeloid Leukemia." Blood 126, no. 23 (2015): 3830. http://dx.doi.org/10.1182/blood.v126.23.3830.3830.
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Abstract Acute Myeloid Leukemia (AML) is a heterogeneous disease with poor overall five-year survival of less than 30%. Current risk stratification is largely based on cytogenetics, combined with information of the most commonly mutated genes in AML (e.g. NPM1, FLT3, DNMT3A). To improve clinical decision making and to increase our understanding of the mechanisms underlying AML it is essential to gain additional information about the mutational landscape of AML. In this prospective study we perform comprehensive Next Generation Sequencing (NGS) to determine the mutational landscape of AML. Starting from September 2014, bone marrow samples, with matched skin biopsies, were collected from all newly diagnosed samples of AML at Skåne University Hospital, Sweden. So far, almost 40 AML samples have undergone whole-exome sequencing (WES) (100X coverage), targeted AML-gene panel sequencing (>100 genes with recurrent mutations in the TCGA AML data set) (400X), RNA-seq and low pass Whole Genome Sequencing (WGS) (1.5X). Additionally, clinical data such as age, treatment response and survival outcome are collected and samples are also cryopreserved for functional follow-up studies. The targeted AML-panel sequencing allows for high coverage data enabling identification of not only common but also rare variants present in subclones, while WES might reveal genes and pathways not previously associated with AML. Low pass WGS enables the detection of cytogenetic alterations, ranging from larger structural rearrangements to fusion gene detection. RNA-seq also makes the detection of fusion genes possible as well as providing global gene expression data. So far our prospective study has identified 22 recurrently mutated genes (with mutations present in >5% of the reads). Out of these, DNMT3A (34%), NPM1 (29%), TET2 (21%), FLT3 (18%) and RUNX1 (18%) were the most commonly mutated genes. The corresponding mutation frequencies in TCGA AML data set are DNMT3A (26%), NPM1 (27%), TET2 (9%), FLT3 (28%) and RUNX1 (10%). More than 70% of the cases carry combinations of mutations in two up to seven of the genes included in our AML panel. Each patient also carries a private combination of unique exomic variants. RNA-seq data confirmed all clinically known fusion genes and principal component analysis revealed that cases with e.g. NPM1 mutations have a uniform gene expression pattern. Although diagnostics has improved over the last years, information of the most commonly mutated genes has not largely improved risk stratification. A plausible explanation is the clonal complexity in AML and the joint risk combination of common and rare variants. NGS-based methods have greatly improved our possibility to detect genetic alterations and comprehensive NGS of AML has the potential to identify mutational patterns that can further improve diagnostics and risk stratification. Disclosures No relevant conflicts of interest to declare.
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Usman, Atif, Nasir Naveed, and Saima Munawar. "Intelligent Anti-Money Laundering Fraud Control Using Graph-Based Machine Learning Model for the Financial Domain." Journal of Cases on Information Technology 25, no. 1 (2023): 1–20. http://dx.doi.org/10.4018/jcit.316665.
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Financial domains are suffering from organized fraudulent activities that are inflicting the world on a larger scale. Basel Anti-Money Laundering (AML) index enlists 146 countries, which are impacted by criminal acts like money laundering, and represents the country's risk level with a notable deteriorating trend over the last five years. Despite AML being a substantially focused area, only a fraction of such activities has been prevented. Because financial data related to this field is concealed, access is limited and protected by regulatory authorities. This paper aims to study a graph-based machine-learning model to identify fraudulent transactions using the financial domain's synthetic dataset (100K nodes, 5.3M edges). Graph-based machine learning with financial datasets resulted in promising 77-79% accuracy with a limited feature set. Even better results can be achieved by enriching the feature vector. This exploration further leads to pattern detection in the graph, which is a step toward AML detection.
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Ramilyeva, I. R., Zh K. Burkitbaev, S. A. Abdrakhmanova, A. A. Turganbekova, D. K. Baimukasheva, and E. B. Zhiburt. "DISTRIBUTION PATTERN FOR HLA SPECIFICITIES IN THE PATIENTS WITH ACUTE MYELOID LEUKEMIA." Medical Immunology (Russia) 21, no. 5 (2019): 965–72. http://dx.doi.org/10.15789/1563-0625-2019-5-965-972.
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The article presents a study on the distribution of gene polymorphisms in the histocompatibility antigens among the patients diagnosed with AML, and healthy donors in the Republic of Kazakhstan, as well as features of the HLA-A*, *B, Cw*, DRB1*, DQB1* distribution among the patients with acute myeloid leukemia (AML). HLA typing and data processing were performed at the Research and Production Center of Transfusiology, Nur-Sultan. A total of 3808 people were examined, including 3621 healthy blood donors and 187 patients diagnosed with AML. Genomic DNA for HLA typing was isolated from peripheral blood leukocytes by proteinase method using columns with silica membrane and using a set of reagents PROTRANS DNA BOX (Protrans, Germany). Typing of HLA-A, B, C, DRB1, DQB1 in the patients and blood donors was performed by polymerase chain reaction using commercial reagent kits from Protrans (PROTRANS HLA- A*/B*/DRB1* Cyclerplate System, PROTRANS HLA-C* Cyclerplate System, PROTRANS HLA-DQB1* Cyclerplate System).HLA-A*31 (OR = 1.8; CI 1.16-2.79; p < 0.01) proved to be more common in the group of patients compared to the control group, which suggesting an association between AML and presence of this antigen. The control group showed an increased frequency of HLA-A*02 antigen (OR = 0.55; CI 0.41-0.75; p < 0.01). This antigen may be, therefore, exert a protective effect in AML development.The studies of major histocompatibility complex which include HLA genes, did significantly expanded the understanding of HLA antigens which may have strong associative links with distinct diseases, and moderately or poorly expressed links in other disorders. Analysis of the literature data showed that myeloid leukemia is characterized by decreased frequency of HLA-B13, B14, B40 antigens, most often determined by antigens B16, Bw 22, B27. In this study, HLA-A*31, B*37 were associated with AML. Phenotypes with antigens HLA-A*02, B*27, C*02, DRB1*01, *04, DQB1*06 have a probable protective effect on the development of this pathology.The study has determined some features of histocompatibility gene distribution in AML patients, detection of HLA-markers that determine the risk or resistance to the occurrence of this disease. We have established characteristic specific markers of HLA system among AML patients in Kazakhstan, which may be associated with higher risk of the disease.
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Zhong, Ling, Yong Qian Jia, Wen Tong Meng, and Xun Ni. "FMS-Like Tyrosine Kinase 3 Internal Tandem Duplication and the Patterns of Its Gene Sequence in 207 Chinese Patients With De Novo Acute Myeloid Leukemia." Archives of Pathology & Laboratory Medicine 136, no. 1 (2012): 84–89. http://dx.doi.org/10.5858/arpa.2010-0700-oa.
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Context.—Constitutive activation of the FMS-like tyrosine kinase 3 (FLT3) receptor tyrosine kinase by internal tandem duplication (ITD) has been researched in patients with de novo acute myeloid leukemia (AML). Objective.—To study the patterns of FLT3-ITD in Chinese patients with AML. Design.—A total of 207 patients with de novo AML were enrolled in the study. Genomic DNA was extracted from peripheral blood and polymerase chain reaction was performed. GeneScan was used to analyze the mutant to wild-type ratio. The sequencing of mutated genes was performed to confirm the mutation types and exclude false positives. Results.—A total of 42 cases (20.3%) were associated with mutations. FLT3-ITD was found equally in AML subtypes M1 to M6. The level of the ITD allele was heterogeneous. GeneScan showed that the mutant to wild-type ratio ranged from 0.03 to 3.78 (median, 0.43). Patients with a high ratio had significantly lower cancer remission rates and shorter survival. They also showed distinct clinical features including higher white blood cell counts and higher CD7 and CD56 expression. The length of the duplicated fragment was 26 to 57 bp (median, 43 bp). Twenty-two cases (52%) had simple tandem duplications, while 20 other cases (48%) had an extra interval of 12 to 30 bp before the tandem duplications. A hexanucleotide consisting of GAAAAG was found exclusively in the intervals. Patients with this GAAAAG interval showed better survival. The ITD to wild-type ratio, gene pattern, and CD7 expression status appear to be independent prognostic indices for patients with AML. Conclusion.—Detection of FLT3 mutation is fast, easy, and inexpensive. The mutant to wild-type ratio is helpful for performing detailed risk stratification. DNA sequence analysis is more precise for confirming and evaluating the mutation pattern.
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Xu, Haosen, Keke Yu, Ming Wei, and Yida Zhu. "Intelligent Anti-Money Laundering Transaction Pattern Recognition System Based on Graph Neural Networks." Journal of AI-Powered Medical Innovations (International online ISSN 3078-1930) 2, no. 1 (2024): 93–108. https://doi.org/10.60087/vol2iisue1.p007.
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This paper presents a new Intelligent Anti-Money Laundering Transaction Pattern Recognition System based on Graph Neural Networks (GNNs). The proposed system addresses the limitations of traditional anti-money laundering (AML) by leveraging the power of image representation and deep learning techniques. We introduce general methods for creating financial networks based on different shapes, including structural and physical. A custom GNN architecture is designed, featuring heterogeneous graph convolution, listening mechanisms, and physical models to capture the exchange patterns. The system uses advanced engineering techniques to extract both local and global features of financial performance. The analysis of the world's big data shows that the best performance of our method, achieved 35.2% Money Laundering Detection Rate (MLDR) in the top 1% of business flag, do better way. The model interpretation is improved by analyzing the SHAP value, providing insight into the decision-making process. Case studies show the system's ability to uncover financial transactions, including deposits from cryptocurrency exchanges and smurfing operations. This research contributes to the advancement of AML practices by introducing more accurate, flexible, and effective solutions for investigating financial crimes in complex financial systems more.
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Haferlach, Torsten, Claudia Haferlach, Alexander Kohlmann, et al. "Microarray Analysis Detects Unique Expression Pattern for NPM1-Mutated AML with Normal Karyotype and Reveals Pathobiological Insights." Blood 110, no. 11 (2007): 2382. http://dx.doi.org/10.1182/blood.v110.11.2382.2382.
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Abstract Recent data indicate that mutations in exon 12 of the nucleophosmin (NPM1) gene characterize a distinct subgroup of adult and pediatric acute myeloid leukemia (AML). AML carrying NPM1 mutations account for about one-third of all adult AML, exhibit distinctive biological and clinical features and show a strong association to AML with normal karyotype (55% mutated). However, the role of NPM1 in leukemogenesis still remains elusive. Here we present data on a cohort of n=66 AML cases with normal karyotype analyzed by high-density whole genome expression microarrays (Affymetrix HG-U133 Plus 2.0). In parallel melting curve analysis was used to assess NPM1 mutational status: 41 cases were characterized as mutated (NPM1+) and 25 cases were unmutated (NPM1−). We first investigated the gene signature that discriminated NPM1+ from NPM1− cases. Genes that were significantly overexpressed comparing NPM1+ against NPM1– cases included a strong homeobox genes signature (HOXA1, HOXA5, HOXA7, HOXA9, HOXA10, HOXA11, HOXB2, HOXB4, HOXB5, HOXB6, HOXB7, MEIS1, and PBX3). A functional analysis (Gene Ontology) revealed a clear association of the group of overexpressed genes with the cell components nucleosome, chromatin, and the nuclear envelope-endoplasmatic reticulum network as well as involvement in the biological processes of nucleosome and chromatin assembly, establishment of protein transport and localization, and Notch signaling pathway. In contrast, the cellular processes completely differed when genes were investigated that were significantly underexpressed in NPM1+ cases compared to NPM1− cases. This group of genes encoded membrane-related proteins (gap junction, intercellular junction, signalosome complex) and proteins involved in cellular morphogenesis and cell communication. The differences in gene expression signatures between NPM1+ and NPM1− cases permit a robust classification approach by gene expression profiling. Support Vector Machine analysis resulted in &gt;92% prediction accuracy of NPM1 mutation status (10-fold cross-validation). The sensitivity was very high for the positive detection of NPM1+ cases (&gt;97%). Using a 100-fold re-sampling approach and splitting the complete data set into a training set (n=44) and testing set (n=22) the following genes were most frequently selected as top discriminatory genes: HOXA5, HOXB4, HOXB5, HOXB6, MEIS1, PBX3, FGFR1, ADAM17, PRICKLE1, and TMPO. Interestingly, the classification was less accurate when also FLT3 internal tandem duplication mutation status was taken into account. The study cohort (n=66) then was distributed as follows: 19 NPM1+/FLT3+, 22 NPM1+/FLT3−, 4 NPM1−/FLT3+, and 21 NPM1−/FLT3− negative cases. Only 14 of 22 (64%) NPM1+/FLT3– cases were correctly predicted, with miscalls falling both into the group of NPM1+/FLT3+ and NPM1−/FLT3− cases. In conclusion, NPM1 mutations are the most frequent mutations in adult AML to date and their central prognostic role is increasingly recognized. Given the fact that they are nearly mutually exclusive with major recurrent genetic abnormalities and that they can be characterized by a distinctive gene expression program these data especially for of NPM1+/FLT3− AML with better outcome may support to classify this as a separate biological subgroup of AML with normal karyotype.
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Martinez, Clara M., Elena Bussaglia, Carmen Hernandez, et al. "Abnormal CD34 Positive Cells Are Present in NPM+ Acute Myeloid Leukemia (AML) Samples at Diagnosis." Blood 110, no. 11 (2007): 4289. http://dx.doi.org/10.1182/blood.v110.11.4289.4289.
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Abstract Nucleophosmin (NPM) gene mutations are the molecular hallmark of a large primary AML subgroup of patients. NPM mutated cases are commonly associated with FLT3-internal tandem duplications which seem to confer an adverse prognostic meaning. Blasts from NPM+ leukemias have been described as CD34 negative. Objective: To assess the presence of CD34 positive populations and their phenotypic profile in NPM mutated samples at diagnosis. To investigate whether there are differences between NPM/FLT3- and those with an associated FLT3-ITD. Patients and methods: Cells from forty-three bone marrow de novo NPM+ AML (26 females/ 17 males) patients enrolled in AML-CETLAM protocols were analyzed at diagnosis by means of multiparameter flow cytometry (MFC). In 18 cases an associated FLT3-ITD was detected (NPM+/FLT3+ group). Immature antigenic markers were investigated by triple combinations of direct conjugated MoAbs to CD15, CD34 and HLA-Dr ; CD34, CD123 and HLA-Dr; CD34, CD33 and CD19; CD15, CD117 and CD45. Minimal residual disease studies (7 cases) were performed in samples in complete remission using flow cytometry and real-time PCR (ABIPRISM 7700). Results: In all the patients we were able to detect an abnormal CD34 population. CD15 coexpression, abnormal light scatter patterns, CD117 clusterization, antigenic overexpression and abnormal CD123 expression were all detected in these cases. NPM+/FLT3+ patients had 2,24% CD34+ cells(0,12%–42%) whereas for NPM+/FLT3- cases the mean CD34+ value was 1,05% (0,1–92%)(n.s.). Sequential samples to test in parallel the abnormal immunophenotype and NPM transcripts by real-time PCR were available in 7 cases (5 NPM+/FLT3+ and 2 NPM+/FLT3-). Relapses were observed in two NPM+/FLT3+ cases (4% and 42% CD34+ cells at diagnosis) and 1 NPM+/FLT3- patient (0.1% CD34+ cells at diagnosis). No definite pattern of relapse could be established given that in some cases NPM rises preceded the detection of phenotypically abnormal cells. Conclusions: NPM mutated AML patients may be mostly categorized as CD34 negative following standard flow-cytometry positivity thresholds. Nevertheless, the constant detection of immature abnormal cells (CD34+CD123+CD117++) may help to identify the leukemic stem-cell pool in NPM+ and FLT3+ leukemias. It remains to be investigated whether the size of this compartment has prognostic relevance in some groups of normal karyotype AML.
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Simonetti, Giorgia, Antonella Padella, Ítalo Faria do Valle, et al. "A Specific Pattern of Somatic Mutations Associates with Poor Prognosis Aneuploid Acute Myeloid Leukemia: Results from the European NGS-PTL Consortium." Blood 126, no. 23 (2015): 3840. http://dx.doi.org/10.1182/blood.v126.23.3840.3840.
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Abstract Aneuploidy causes a proliferative disadvantage, mitotic and proteotoxic stress in non-malignant cells ( Torres et al. Science 2007). Chromosome gain or loss, which is the hallmark of aneuploidy, is a relatively common event in Acute Myeloid Leukemia (AML). About 10% of adult AML display isolated trisomy 8, 11, 13, 21 (Farag et al. IJO 2002), or either an isolated autosomal monosomy or monosomal karyotype (Breems et al. JCO 2008). This evidence suggests that tumor-specific mechanisms cooperate to overcome the unfitness barrier and maintain aneuploidy. However, the molecular bases of aneuploid AML are incompletely understood. We analyzed a cohort of 166 cytogenetically-characterized AML patients (80 aneuploid (A-) and 86 euploid (E-)) treated at Seràgnoli Institute (Bologna). Aneuploidy was significantly associated with poor overall survival (median survival: 13 and 26 months in A-AML and E-AML respectively; p=.006, Fig.1). To identify AML-specific alterations having a causative and/or tolerogenic role towards aneuploidy, we integrated high-throughput genomic and transcriptomic analyses. We performed 100 bp paired-end whole exome sequencing (WES, Illumina Hiseq2000) of 70 samples from our A-AML and E-AML cohort of 166 patients. Variants where called with MuTect or GATK for single nucleotide variant and indels detection, respectively. AML samples were genotyped by CytoScan HD Array (Affymetrix). Gene expression profiling (GEP) was also conducted on bone marrow cells from 24 A-AML, 33 E-AML (≥80% blasts) and 7 healthy controls (HTA 2.0, Affymetrix). We detected a significantly higher mutation load in A-AML compared with E-AML (median number of variants: 31 and 15, p=.04) which was interestingly unrelated to patients' age (median age: 63.5 years in A-AML and 62 years in E-AML, Xie et al, Nat. Med. 2014). C>A and C>T substitutions, which are likely mediated by endogenous 5mCdeamination, were the most frequent alterations (Alexandrov et al. Nat. 2013). However, aneuploidy associated with an increased variability in terms of mutational signatures, with the majority of A-AML displaying 3 or more signatures compared to few E-AML cases (p=.04). WES analysis also revealed a specific pattern of somatic mutations in A-AML. A-AML had a lower number of mutations in signaling genes (p=.04), while being enriched for alterations in cell cycle genes (p=.01) compared with E-AML. The mutated genes were involved in different cell cycle phases, including DNA replication (MCM6, PURB, SSRP1), centrosome dynamics (CEP250, SAC3D1, HEPACAM2, CCP110), chromosome segregation (NUSAP1, ESPL1, TRIOBP), mitotic checkpoint (ANAPC7, FAM64A) and regulation (CDK9, MELK, ZBTB17, FOXN3, PPM1D, USP2). Moreover, genomic deletion of cell cycle-related genes was frequently detected in A-AML. Notably, ESPL1 which associated with aneuploidy, chromosome instability and DNA damage in mammary tumors (Mukherjee et al. Oncogene 2014) was mutated and also upregulated in A-AML compared with E-AML (p=.01), the latter showing expression levels comparable to controls. Among the top-ranked genes differentially expressed between A-AML and E-AML, we identified a specific signature characterized by increased CDC20 and UBE2C and reduced RAD50 and ATR in A-AML (p<.001), which has been previously linked to defects in chromosome number. Additional mutations targeting DNA damage and repair pathways were identified in A-AML, including TP53 mutations, which account for 15% of cases. Moreover, A-AML showed a significant upregulation of a KRAS transcriptional signature and downregulation of FANCL- and TP53-related signatures, irrespective of TP53 mutational status. Our data show a link between aneuploidy and genomic instability in AML. Deregulation of the cell cycle machinery, DNA damage and repair checkpoints either through mutations, copy number and transcriptomic alterations is a hallmark of A-AML. The results define specific genomic and transcriptomic signatures that cooperate with leukemogenic pathways, as KRAS signaling, to the development of the aggressive phenotype of A-AML and suggest that a number of A-AML patients may benefit frompharmacological reactivation of TP53pathway (e.g. MDM2 inhibitor, clinical trial NP28679). Supported by: FP7 NGS-PTL project, ELN, AIL, AIRC, PRIN, progetto Regione-Università 2010-12 GS & AP: equal contribution Disclosures Soverini: Novartis, Briston-Myers Squibb, ARIAD: Consultancy. Cavo:JANSSEN, CELGENE, AMGEN: Consultancy. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Martinelli:MSD: Consultancy; BMS: Speakers Bureau; Roche: Consultancy; ARIAD: Consultancy; Novartis: Speakers Bureau; Pfizer: Consultancy.
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Fauser, A. A., L. Kraut, G. Trebeljahr, B. Thiele, N. Gulzad, and E. Roemer. "Complete remission after induction therapy of donor cell derived secondary AML in a CML transplant patient." Journal of Clinical Oncology 24, no. 18_suppl (2006): 16523. http://dx.doi.org/10.1200/jco.2006.24.18_suppl.16523.
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16523 Background: Malignant diseases arising from donor derived cells are an exceptionally rare condition with approximately 20 cases only reported in the literature. We report on diagnosis and salvage chemotherapy of a CML transplant patient relapsing with donor cell derived secondary MDS resp. AML. Methods: Chimerism analysis were performed by quantitative short tandem repeat (STR) DNA detection in bone marrow as well as in peripheral blood, detection limit 0.5%. Standard procedures were used for all other methods. Results: In a female patient transplanted with a HLA-DRB1 mismatched female unrelated donor in July 1998 for Ph+ CML, secondary MDS was diagnosed in March 2005 transforming into secondary AML in June 2005. The results from chimerism analysis after diagnosis of sMDS resp. sAML in this transplant patient, i.e. no detection of the STR pattern of the recipient, clearly confirmed a donor cell derived sMDS resp. sAML almost 7 years after allogeneic BMT. Cytogenetics showed a normal female karyotype, and no molecular aberrations were detected by FISH resp. PCR. After diagnosis of sAML the patient received 2 induction courses consisting of FLAG-Ida and MTC regimen, respectively. Bone marrow analysis after the first and second course revealed no evidence of blasts whereas the MDS continued to be detectable. Cytogenetics during and after reinduction therapy were normal. Repeated chimerism analysis showed complete donor chimerism (> 99.5%). Thus, a complete remission of donor cell derived sAML using conventional induction chemotherapy was achieved. This remission is now lasting 6 months without intensification. Medical evaluation of the donor did not reveal any disturbed or malignant hematological disorder. Conclusions: This is an interesting case of rare donor cell derived leukemia and its successful treatment. The question arises with regard to altered stromal factors in the bone marrow of the patient as potential cause of the disease. No significant financial relationships to disclose.
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Beyá, Marina Díaz, Alfons Navarro, Tania Díaz, et al. "A Distinctive MicroRNA Signature Characterizes Acute Myeloid Leukemia with Translocation (8;16)(p11;p13) and MYST3-CREBBP Rearrangement." Blood 116, no. 21 (2010): 230. http://dx.doi.org/10.1182/blood.v116.21.230.230.
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Abstract Abstract 230 Acute myeloid leukemia (AML) with t(8;16)(p11;p13) and MYST3-CREBBP rearrangement [t(8;16) AML] is an infrequent leukemia subtype with specific clinical and biological characteristics. Although a characteristic gene expression pattern has been identified with t(8;16) AML, to date, the microRNA (miRNA) expression pattern of this subtype has not been described. The main objective of this study was to analyze the expression pattern of mature miRNAs in patients with t(8;16) AML and compare it with other well defined AML subtypes. We have analyzed samples from 117 AML patients and three CD34+ bone marrow specimens from healthy donors. In addition to five cases of t(8;16) AML, samples from nine other AML subtypes were included.The expression of 670 mature miRNAs was analyzed using TaqMan Human MicroRNA Arrays v2.0 (Applied Biosystems) in an ABI 7900 HT sequence detection system. miRNA expression data was analyzed by the 2−ΔΔCt method, using RNU48 as endogenous control. Statistical analyses were performed with TiGR MultiExperiment Viewer and R software. In order to identify miRNA targets, we used the RmiR package (Bioconductor) to cross-correlate the miRNA expression data from the present study with our previous findings on gene expression in the same patient samples (Camos M et al, Cancer Res 2006), based on the predicted targets from TargetScan and Pictar databases. The unsupervised hierarchical cluster analysis showed well-differentiated groups correlating with cytogenetic and molecular categories. Interestingly, all t(8;16) AML patients were grouped in an independent cluster. Supervised analysis by means of t-test and SAM analysis revealed a distinctive 108-miRNA signature in t(8;16) AML patients. All 108 miRNAs were downregulated in t(8;16) AML in comparison to the other subtypes, including all the miRNAs of cluster miR-17-92 and its paralogs miR-106b-25 and 106a–363, as well as miR-29b.Since 14% of the 108 miRNAs in the signature, including miR17-92 and its paralogs, are transcriptionally activated by c-Myc, we then examined c-Myc mRNA analysis by RT-QPCR. Expression of c-Myc in our t(8;16) AML patients was significantly downregulated (p=0.02).miR-29b, also downregulated in our t(8;16) AML patients, is known to target DNA methyltransferase (DNMT) genes, which play a key role in the regulation of DNA methylation through CpG methylation. Since 8% of the 108 miRNAs in our signature are located in CpG islands and 29% are intronic miRNAs contained in genes with a CpG island in their promoter region, we speculated that DNMT activity could also be a factor in the overall downregulation of the miRNAs in the signature.When miRNA data from the present study were cross-correlated with our previous findings on mRNA expression in the same patients, we found an inverse correlation of miR-130a and miR-130b with HOXA3 (r2= -0.5; -0.8; respectively), of miR-1 and miR-23b with MEIS1 (r2= -0.62; -0.62; respectively) and of miR-15b, miR-195 and miR-218 with RET (r2= -0.92;-0.58;-0.87; respectively). In summary, t(8;16) AML exhibits a distinctive 108-miRNA signature, with an overall downregulated profile, which may be partially explained by c-Myc downregulation. Moreover, we have identified potential target genes of these miRNAs which had previously been shown to be characteristic of this AML subtype. Our findings thus contribute some insight into the biological profile of t(8;16) AML and can provide a greater understanding of genetic and epigenetic abnormalities in t(8;16) AML. Disclosures: No relevant conflicts of interest to declare.
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Nafisat Temilade Popoola and Felix Adebayo Bakare. "Advanced computational forecasting techniques to strengthen risk prediction, pattern recognition, and compliance strategies." International Journal of Science and Research Archive 12, no. 2 (2024): 3033–54. https://doi.org/10.30574/ijsra.2024.12.2.1412.
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In an era defined by data-driven decision-making, advanced computational forecasting techniques have emerged as powerful tools for strengthening risk prediction, pattern recognition, and compliance strategies. These techniques leverage artificial intelligence (AI), machine learning (ML), and big data analytics to enhance accuracy, efficiency, and reliability in risk assessment across diverse industries. Traditional risk prediction models often rely on historical data and statistical methods, which, while effective, struggle to capture complex, non-linear patterns in evolving datasets. Advanced computational techniques, such as deep learning, ensemble learning, and reinforcement learning, have significantly improved predictive capabilities by identifying intricate correlations and anomalies in vast datasets. Pattern recognition plays a crucial role in cybersecurity, fraud detection, and financial risk management, where real-time anomaly detection enables organizations to preemptively mitigate threats. Predictive analytics models integrated with neural networks and natural language processing (NLP) have further revolutionized compliance strategies, ensuring adherence to regulatory frameworks and minimizing operational risks. In financial institutions, computational forecasting optimizes credit risk assessment and anti-money laundering (AML) monitoring, while in healthcare, it enhances disease outbreak predictions and patient care strategies. Despite these advancements, challenges such as algorithmic biases, data privacy concerns, and interpretability issues remain. Regulatory bodies are increasingly scrutinizing AI-driven decision systems to ensure transparency, fairness, and accountability. This study provides a comprehensive analysis of the latest computational forecasting techniques, their applications in risk management, and the evolving regulatory landscape. By addressing existing challenges and optimizing these techniques, industries can leverage AI-driven forecasting to enhance resilience, mitigate risks, and maintain regulatory compliance in an increasingly complex digital ecosystem.
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Huh, Jungwon, Chul Won Jung, Hyeoung-Joon Kim, et al. "Different Characteristics Identified by Single Nucleotide Polymorphism Array Analsysis in Leukemia Suggest the Need for Different Application Strategics Depending On Disease Category." Blood 120, no. 21 (2012): 2490. http://dx.doi.org/10.1182/blood.v120.21.2490.2490.
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Abstract Abstract 2490 Background: High resolution SNP-A (single nucleotide polymorphism array) based karyotyping has become one of the most useful tools to investigate complex genomic lesions in leukemic cells. SNP-A has the potential to be applied as a diagnostic tool and as a surrogate for prognostic stratification in leukemias. As such, SNP-A should be a valuable asset in the diagnostic flow in clinical cytogenetic setting. However, due to limited laboratory and health care resources, its application needs to be decided carefully with a reasonable guideline. The present study investigated the value of SNP-A as an array based karyotyping tool in 469 cases with leukemias/MDS. The detection rate of SNP-A based karyotyping in combination with metaphase cytogenetics (MC) was assessed, with the goal of proposing a practical approach for clinical applications of SNP-A in leukemias/MDS. Material and Methods: Four hundred sixty nine patients with diverse subtypes of leukemia were included; 133 AML with normal karyotype (AML-NK), 98 AML with core binding factor rearrangement (CBF-AML), 61 MDS including chronic myelomonocytic leukemia, 118 CML in chronic phase (CML-CP), and 59 ALL (B-lineage). The Genome-Wide Human SNP 6.0 Array (Affymetrix, Santa Clara, CA, USA) was performed. Results: In the 469 patients, the detection rate was 20%(96/469) by MC alone, 33%(156/469) by SNP-A alone, and 39%(184/469) by combined use of SNP-A and MC. According to subtype of leukemias/MDS, the detection rate identified by SNP-A was different and the incidence of CN-LOH varied. In addition, each leukemia subtype showed a different pattern in terms of their types, sizes, and locations of copy number alterations and CN-LOH lesions. SNP-A detected chromosomal lesions not identified by MC, especially in AML-NK with a detection rate of 32%(43/133). In CBF-AML, the detection rates by MC alone and combined use of MC and SNP-A were 29%(28/98) and 40%(39/98), respectively. AML-NK showed more frequent CN-LOH (23%,31/133) than CBF-AML (3%,3/98). In AML-NK, loss of lesions occurred predominantly on 17p, 17q, or 7q. In AML-CBF, 8q or 9q was the common site of loss lesions. Trisomy 8 and CN-LOH of 13q were common lesions in both AML-NK and AML-CBF. As for MDS, t he detection was improved to 54%(33/61) by combined SNP-A and MC, compared with 39%(24/61) using MC alone. MDS showed frequent CN-LOH (25%,15/61) on 11q or 14q. Consistent with MC, loss lesions were frequently found on 5q, 7q, or 20q. In one patient with del(9q) as a sole abnormality, CN-LOH of 7q (26 Mb) was found by SNP-A. In ALL, detection was improved to 88%(51/59) by using combined SNP-A and MC, compared with 63%(37/59) by MC alone and CN-LOH was found in 20%(12/59) of patients. Concerning the size of abnormal lesions, the median size of loss in ALL was smaller than those of other leukemia subtypes. The size of focal deletions of IKZF 1, ETV 6, and CDKN 2A was 106–282 kb, 370 kb-4.5 Mb, and 115 kb-2.1 Mb, respectively. Furthermore, in ALL, aneuploidy abnormalities were accurately identified by SNP-A. ALL case with normal karyotype by MC showed gain or loss of several whole chromosomes by SNP-A, suggesting presence of hyperdiploidy clones or hypodiploidy clones. In particular, in a case showing chromosome numbers of 46–60 by MC, SNP-A revealed two kinds of abnormal lesions; CN-LOH and gain (tetrasomy) on several whole chromosomes, consistent with doubling of a near-haploid clone. In CML, t he detection rates of MC alone and combined use of MC and SNP-A was 3%(4/118) and 24%(28/118), respectively. Among abnormal lesions identified by SNP-A, 90% (35/39) were losses; 83%(29/35) were submicroscopic 9q34 or/and 22q11.2 deletions adjacent to the t(9;22) breakpoint. CN-LOH was found only in 2%(2/118) in CML-CP. Conclusion: The present study shows that SNP-A is useful to improve the detection rate of genomic lesions in diverse leukemia subtypes. The decision to perform SNP-A in addition to MC and/or FISH should be made according to the disease category of each leukemia subtype. In AML, it would be better that SNP-A is used depending on the karyotypic results by MC, while routine SNP-A appears to be an efficient screening method in MDS and ALL and the benefit is minimal in CML A consensus needs to be reached between laboratories as to the practical strategies for the clinical application of SNP-A. Disclosures: No relevant conflicts of interest to declare.
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Lahortiga, Idoya, Iria Vazquez, Nerea Marcotegui, et al. "GATA2 Is Overexpressed in 46% of Patients with AML and Normal Karyotype. The Mutational Pattern FLT3-ITD/GATA2/WT1 Could Define a Group of Patients with Normal Karyotype and AML-M1 Subtype." Blood 106, no. 11 (2005): 2378. http://dx.doi.org/10.1182/blood.v106.11.2378.2378.
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Abstract The presence of recurrent cytogenetic aberrations in AML has diagnostic and prognostic value; however, more than 40% of patients have a normal karyotype. Although the presence of FLT3 mutations, or overexpression (OE) of WT1, EVI1 or BAALC, have been reported to discriminate in this group those with a worse prognosis, more molecular markers are necessary to achieve treatment stratification in this heterogeneous group. The mice homolog of EVI1 (3q26) has been reported to have a role in the HSC proliferation through Gata2 expression, a downstream target gene of the Evi1 oncogene. Therefore, GATA2 (3q21), a transcription factor with a relevant role in hematopoiesis, could be a candidate gene in the leukemogenic transformation of AML. Our aim was to determinate the incidence of GATA2 OE in AML, and its association with other well-defined prognostic factors. We analyzed 7 cell lines and 192 samples of AML patients at diagnosis: 61 were in the favorable prognosis group, 82 in the intermediate (68 with normal karyotype) and 56 in the unfavorable (27 with 3q rearrangements). Expression of GATA2, MDS1/EVI1, EVI1 and WT1 were measured by a RTQ-PCR Taqman assay (Applied Biosystems). Mutations of FLT3 (ITD and D835) were also analyzed. Gene expression arrays were performed with the Human 19K Oligo Array (Center for Applied Genomics, University of New Jersey). We detected a different expression profile among the 3 prognosis groups: 42% and 12% of all AML cases had GATA2 and EVI1 OE respectively, and the frequency increased in the poorer prognosis groups (Table 1). GATA2 OE was detected in 46% of cases with normal karyotype, and was more frequent among samples with FLT3-ITD (68% vs 34%; p=0.0021). Among AML-M1 cases, the number of samples with FLT3-ITD and GATA2 OE was twice than in samples without FLT3 mutations (78% vs 39%). All patients with AML-M1, FLT3-ITD and GATA2 OE had WT1 OE. Gene expression array technology was performed to compare 3 groups of AML-M1: I) FLT3 no mutated and GATA2 OE; II) FLT3 no mutated and GATA2 no OE; and III) FLT3-ITD and GATA2 OE. Unsupervised analysis clearly classified these 3 groups, suggesting they could be prognostically relevant AML subgroups, and allowing the detection of 38 candidate genes that were differentially expressed. Our results show that GATA2 OE is a common event in AML cases with normal karyotype (46%). The mutational pattern FLT3-ITD/GATA2/WT1 could define a subgroup of patients with normal karyotype and AML-M1, with a different expression pattern. Incidence of the OE of GATA2 and EVI1 in 199 patients with AML GATA2 EVI1 Frequency p value Frequency p value FG: favorable group IG: intermediate group UG: unfavorable group ns: no significative. FG vs IG 26% (16/61) vs 40% (33/82) ns 0% (0/61) vs 4% (3/82) ns FG vs UG 26% (16/61) vs 57% (32/56) 0.01 0% (0/61) vs 38% (21/56) 0.001 IG vs UG 40 (33/82) vs 57% (32/56) ns 4% (3/82) vs 38% (21/56) 0.001
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Kolomietz, Elena, Jaudah Al-Maghrabi, Shawn Brennan, et al. "Primary chromosomal rearrangements of leukemia are frequently accompanied by extensive submicroscopic deletions and may lead to altered prognosis." Blood 97, no. 11 (2001): 3581–88. http://dx.doi.org/10.1182/blood.v97.11.3581.
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BCR/ABL fluorescent in situ hybridization study of chronic myeloid leukemia (CML) and Philadelphia+(Ph+) acute lymphoid leukemia (ALL) indicated that approximately 9% of patients exhibited an atypical hybridization pattern consistent with a submicroscopic deletion of the 5′ region ofABL and the 3′ region of the BCR genes on the 9q+ chromosome. The CML patients with deletions had a shorter survival time and a high relapse rate following bone marrow transplant. Since deletions are associated with both Ph+CML and ALL, it seemed probable that other leukemia-associated genomic rearrangements may also have submicroscopic deletions. This hypothesis was confirmed by the detection of deletions of the 3′ regions of theCBFB and the MLL genes in AML M4 patients with inv(16) and in patients with ALL and AML associated withMLL gene translocations, respectively. In contrast, analysis of the AML M3 group of patients and AML M2 showed that similar large deletions were not frequently associated with the t(15;17) or t(8;21) translocations. Analysis of sequence data from each of the breakpoint regions suggested that large submicroscopic deletions occur in regions with a high overall density of Alu sequence repeats. These findings are the first to show that the process of deletion formation is not disease specific in leukemia and also implicate that the presence of repetitive DNA in the vicinity of breakpoint regions may facilitate the generation of submicroscopic deletions. Such deletions could lead to the loss of one or more genes, and the associated haploinsufficiency may result in the observed differences in clinical behavior.
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Ahlgren, Louise, Mattias Pilheden, Helena Sturesson, et al. "The Relapse Mechanisms and Genomic Landscape Differ in KMT2A-r Pediatric Leukemia in Relation to Relapse Time." Blood 142, Supplement 1 (2023): 2978. http://dx.doi.org/10.1182/blood-2023-185044.
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The genomic landscape and mechanisms driving relapse in KMT2A-rearranged ( KMT2A-r) infant and childhood acute lymphoblastic (ALL) and acute myeloid leukemia (AML) are not completely understood. We therefore studied 36 KMT2A-r ALL (n=19) and AML (n=17) patients of which 25 relapsed and 11 remained in remission. Twenty diagnose-relapse-germline trios and 5 multiple relapse samples were analyzed by whole genome (WGS) and whole exome sequencing (WES) and 30 patients longitudinally by using patient-specific mutations identified by WGS/WES, including the KMT2A-r (average coverage 3300X). The mutational burden increased from diagnosis to relapse and relapse evolved through branching evolution. Relapse was seeded by multiple diagnostic clones in 56%, by a single sweeping clone detected at diagnosis in 22%, and by a single sweeping clone not detected at diagnosis in 22%. Notably, the evolutionary patterns correlated to relapse time, where multiple diagnostic clones seeding relapse were connected to an earlier relapse with all very early relapse ALL (3/3, relapse &lt;9 months from diagnosis) and half of the early AML relapse showing this pattern (2/4, relapse &lt;1 year in complete remission, CR1). By contrast, later relapse was connected to a sweeping clone at relapse with 67% of early relapse ALL (&gt;9 months from diagnosis) and 40% of late relapse AML (&gt;1 year in CR1) showing this pattern. Pathway analysis showed that cell cycle genes, glucocorticoid signalling, purine metabolism, mismatch repair, and B-cell differentiation, were enriched in early relapse ALL (83%, 5/6) and included TP53, CREBBP, NT5C2 PMS2, PRPS2, NR3C1, IKZF1, with none of the very early relapse infant ALL harboring such alterations (n=4). Further, TP53 and IKZF1alterationsco-occurred (n=4/4). These results were validated in public data sets of 98 KMT2A-r ALL infants (n=84) and children (n=14) at diagnosis and relapse (n=24) and showed that 50% of early relapse ALL, and none of the 8 very early relapse ALL, had such alterations. Ultra-deep sequencing did not detect the CREBBP, NT5C2, PRPS2or TP53 mutations at diagnosis and manual inspection of the WGS reads failed to detect the PMS2 and NR3C1 deletions. In AML, TP53 and CCND3 alterations were maintained, and gain of WT1 was seen in late relapse AML. Signalling mutations were the most common type of mutations at diagnosis (64%) and relapse (56%) and the frequency was similar in patients that remained in remission and in those that relapsed (55% versus 60%). One infant ALL and four AML patients had multiple relapses, allowing us to study how the genetic landscape evolved across consecutive relapses. This showed a stepwise replacement of clones during treatment in agreement with a fitter clone that evolves under chemotherapeutic selective pressure. Longitudinal analysis allowed sensitive detection of residual leukemia cells and showed that the relapse clone could be detected at diagnosis in 64% of patients. Further, infants with &gt;10% of molecularly detectable leukemia cells after induction therapy, had a high risk of a very early relapse. Ultra-deep sequencing allowed detection of the relapse clone up to 4 months before relapse. In 11 of the 30 patients (3 remission and 8 relapse), low-frequency KMT2A-fusion positive leukemic cells were found at remission outside of the MRD time points. Our longitudinal data also provided unique insights into clonal response to treatment by showing that 1) a change in therapy can favour the eradication of one clone and expansion of another, 2) a clone that initially was the most sensitive clone to therapy, was the one that eventually caused relapse, and 3), a diagnostic clone can be undetectable for a long time before expanding to cause relapse, suggesting that molecular monitoring with personal mutations is a powerful tool to follow response to therapy. These results provide new biological insights into the relapse mechanisms in KMT2A-r leukemia. The data shows different clonal evolution patters depending on when in time the patient relapsed, with very early relapse ALL being seeded by multiple diagnostic subclones and a paucity of acquired genetic alterations at relapse. By contrast, early relapse ALL was characterized by a single diagnostic clone seeding relapse by a clonal sweep along with acquired mutations in chemoresistance-associated genes. To validate and extend these findings, we are currently analyzing 11 additional infant relapse samples with WGS.
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Calabrese, Chiara, Gian Matteo Pica, Enrico Bracco, et al. "Detection of an Unbalanced Ratio Between WT1 Isoforms in Acute Myeloid Leukemia and Its Correlation with Molecular Abnormalities." Blood 120, no. 21 (2012): 2500. http://dx.doi.org/10.1182/blood.v120.21.2500.2500.
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Abstract Abstract 2500 Wilms' tumor gene 1 (WT1) is a tumor suppressor gene coding for a zinc finger transcriptional factor which was found overexpressed in acute myeloid leukemias (AML). In AML WT1 functions as an oncogene rather than a tumor suppressor. This may depend on the unbalanced expression of its different isoforms derived from alternative splicing in exon 5 (EX5+/−) or the presence of KTS in exon 9 ( KTS+/KTS-). In normal hematopoietic cells the physiological ratio between KTS+/KTS- isoforms is 1:1. KTS are inserted between the third and fourth zinc fingers thus influencing the DNA binding and therefore the transcriptional activity. The aim of the study was to investigate the possibility of an unbalanced ratio between the isoforms thus leading to the disruption of WT1 function. In addition we correlated KTS+ isoform with clinical and biological features. Methods: we analyzed KTS+/KTS- isoforms in 120 AML patients ( 102 BM samples, 18 PB), 63 chronic phase CML patients (48 BM and 15 PB). In addition we evaluated 5 samples of selected blast cells from AML patients. In a subset of 38 adult acute myeloid leukemia (AML) patients we measured all WT1 isoforms (A[EX5 -/KTS -], B[+/−], C[-/+] and D[+/+]. Quantitative PCR was used for the WT1 isoforms detection and measurement. Results: KTS+ isoform is significantly overexpressed with a ratio KTS+/KTS− 2:1 in both AML and CML patients and it is confirmed in blast cell samples. In the subgroup of patients analyzed for the expression pattern of all WT1 isoforms we detected a significant overexpression of isoform D (EX5+/KTS+) (median value 821) while the isoform A (EX5−/KTS−) is the less represented ( median value 303). In this subgroup the expression of KTS+ isoforms (B+D) is higher than KTS- ( A+C) with a median value of 642 compared to 421. The ratio of WT1 isoforms was not significantly different among FAB subgroups or according to cytogenetic risk, FLT3 (fms-like tyrosine kinase receptor-3) internal tandem duplication or exon 17 mutation, NPM (nucleophosmin) gene mutations and BAALC (brain and acute leukemia, cytoplasmic) gene expression. Conclusion: Although WT1 is overexpressed in AML and CML patients it lacks its typical tumor suppressor function. In this study we have demonstrated the overexpression of WT1 KTS+ isoforms. In addition we have previously demonstrated that KTS+ isoform does not have transcriptional activity since it is mainly localized within the cytoplasm. In conclusion, WT1 is overexpressed in AML but the unbalanced ratio between the isoforms may probably influence the transcriptional activity. Any possible correlation between the unbalanced ratio of WT1 isoforms and clinical outcome requires further prospective studies to be established. Disclosures: No relevant conflicts of interest to declare.
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Elizabeth Kuukua Woode Amoako, Victor Boateng, Ola Ajay, Tobias Kwame Adukpo, and Nicholas Mensah. "Exploring the role of Machine Learning and Deep Learning in Anti-Money Laundering (AML) strategies within U.S. Financial Industry: A systematic review of implementation, effectiveness, and challenges." Finance & Accounting Research Journal 7, no. 1 (2025): 22–36. https://doi.org/10.51594/farj.v7i1.1808.
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As the U.S. financial sector confronts evolving threats from financial crimes, the integration of Machine Learning (ML) and Deep Learning (DL) into Anti-Money Laundering (AML) strategies has become imperative. This paper explores the role of ML and DL technologies in enhancing AML frameworks to identify, mitigate, and prevent money laundering activities. The paper begins by analyzing prevalent money laundering schemes and the methods used by criminals to bypass traditional AML controls. The study underscores the importance of educating and training financial institution personnel to ensure the effective implementation of AML strategies powered by ML and DL. The findings revealed that a culture of awareness and accountability is vital for managing risks associated with financial crimes. Furthermore, the paper highlights the value of collaboration and information-sharing between financial institutions, regulatory bodies, and technology providers. Industry partnerships, public-private initiatives, and shared threat intelligence are identified as key components in strengthening AML defenses. This research also examines the transformative potential of ML and DL in AML. It shows how these technologies enhance pattern recognition, anomaly detection, and decision-making processes, allowing financial institutions to stay ahead of evolving money laundering tactics. Moreover, the dynamic and self-learning capabilities of ML and DL models enable continuous adaptation to new risks. Through adaptation of a vigilant, collaborative, and technology-driven approach, U.S. financial institutions can leverage ML and DL to enhance AML frameworks, safeguard consumer trust, and protect the integrity of the financial system. Keywords: Money Laundering, Financial Industry, Deep Learning, USA
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Watson, Caroline J., Sophia Apostolidou, Usha Menon, and Jamie R. Blundell. "Tracing the Evolution of Clonal Hematopoiesis to AML Using Longitudinal Pre-Diagnosis Blood Samples." Blood 138, Supplement 1 (2021): 599. http://dx.doi.org/10.1182/blood-2021-147503.
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Abstract The acquisition of somatic mutations in hematopoietic stem and progenitor cells (HSPCs) is increasingly common with age (`clonal hematopoiesis'). If sequential acquisition and clonal expansion of mutations occurs, progression to Acute Myeloid Leukemia (AML) can occur. While the mutational landscape of clonal hematopoiesis antecedent to AML development has been well-defined (Abelson et al. 2018, Desai et al. 2018), the timing of acquisition and growth dynamics of these high-risk mutations remain largely unknown. At what age are these mutations acquired? Are the fitness effects (growth rates) conferred by specific mutations predictable from person-to-person and how do fitness effects change with additive mutations? Are the clonal dynamics that precede AML development characterised by strong competition between clones (clonal interference)? To answer these questions, we identified 220 women from the United Kingdom Collaborative Trial of Ovarian Cancer Screening (UKCTOCS) who were cancer-free at enrolment but subsequently developed AML during the &gt;12 years follow-up. 50 of these women had annual blood samples collected at multiple time-points pre-AML diagnosis (mean: 5 time-points, range: 2-11). Deep error-corrected duplex sequencing, with a variant allele frequency (VAF) detection limit of 0.1%, was performed on peripheral blood DNA from these women, as well as from age- and timepoint-matched controls who remained blood cancer free. A custom designed next-generation sequencing (NGS) panel was used to enable detection of mutations in 34 clonal hematopoiesis/AML-associated genes, genome-wide mosaic chromosomal alterations (mCAs) and AML-associated translocations. Having samples from multiple timepoints enabled the fitness effects (growth rates) of mutations to be calculated, as well as the additive effect of further mutations. These growth rates, in combination with insights from evolutionary theory, allowed the acquisition time of many mutations to be estimated, with initiating driver mutations often arising in the first 2 decades of life in the pre-AML cases. Growth trajectory dynamics of co-occurring mutations enabled the clonal composition to be inferred in many instances and revealed linear evolution of successive mutations in some pre-AML cases, but a branching pattern with clear evidence of clonal interference in others. Specific variants, which we have previously identified as 'highly fit' in clonal hematopoiesis (Watson et al. 2020), were significantly enriched in pre-AML cases compared to controls and were often detectable at VAFs &gt;10% more than 5 years pre-diagnosis. NPM1 mutations, which characteristically occur `late' in AML development, could be detected as early as 2 years pre-diagnosis, highlighting the benefit afforded by error-corrected low VAF variant calling, particularly in high-risk individuals. Our findings, exploiting longitudinal blood samples collected pre-AML combined with an integrated assessment of multiple types of genetic changes, reveal key insights into the evolutionary dynamics of mutations in the years preceding AML development. Understanding which features distinguish pre-malignant from benign clonal evolution is key for risk stratification of individuals with clonal hematopoiesis to allow rational monitoring and identification of individuals that may benefit from early intervention studies. Figure 1 Figure 1. Disclosures Watson: Johnson & Johnson: Consultancy; Inivata: Consultancy. Menon: Abcodia Ltd: Current holder of individual stocks in a privately-held company. Blundell: Johnson & Johnson: Consultancy; Inivata: Consultancy.
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Imamura, Toshihiko, Kenichi Sakamoto, Norio Shiba, et al. "Negative CD19 Expression Is Associated with Inferior Relapse-Free Survival in RUNX1-RUNX1T1-Positive Acute Myeloid Leukemia; The Japanese Pediatric Leukemia/Lymphoma Study Group Experience from the AML-05 Study." Blood 132, Supplement 1 (2018): 2810. http://dx.doi.org/10.1182/blood-2018-99-115553.
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Abstract Background: Acute myeloid leukemia (AML) with RUNX1-RUNX1T1 and CBFB-MYH11 have been recognized as core-binding factor (CBF) AML accounting for 25% of the pediatric AML patients. CBF-AML patients have been considered to have a good prognosis, but 30 - 50 % of the RUNX1-RUNX1T1-positive AML patients experience relapse. This finding suggests that some population of them have risk factors associated with poor outcome. Previous studies revealed that KIT activating mutation as a predictor of poor outcome. In addition, previous studies of relatively small number of patients also revealed that CD56 positivity or CD19 negativity were also poor prognostic factors. However, the relationship between KIT activating mutation and specific pattern of cell surface antigens has not been fully investigated. Aim and methods: We performed a retrospective analysis of RUNX1-RUNX1T1-positive AML patients treated in the Japanese Pediatric Leukemia/Lymphoma Study Group (JPLSG) AML-05 protocol to determine risk factors of relapse using the integration of data including pattern of cell surface markers on leukemic cell at diagnosis, genetic abnormalities, and clinical characteristics. Flow cytometric analysis of immunophenotyping was performed in the central laboratories using same panel of antibodies. Conventional cytogenetic analysis using G-banding was performed as part of the routine work-up. Molecular study using quantitative RT-PCR for the detection of RUNX1-RUNX1T1 and PCR for the detection of FLT3-ITD was also performed as part of the routine work-up. Screening of mutation of 8 genes, such as NRAS, KRAS, KIT, WT1, C/EBPA, ASXL1, ASXL2, and CSF3R, was performed by genomic PCR and Sanger sequencing. A total of 106 AML patients enrolled in the JPLSG AML-05 study were RUNX1-RUNX1T1-positive AML, but we could not obtain the data of cell surface marker in 6 of them. Thus, we analyzed 100/106 (94.3%) patients with RUNX1-RUNX1T1-positive AML. Statistical analysis was performed by Kaplan-Meier method with log-rank test. A Cox proportional hazards model was used to determine risk factors for survival and relapse. Results are reported as adjusted odds ratios with 95% confidence intervals. Statistical significance was set at p < 0.05. Results: In entire study population, 8 of the 100 patients died and 24 of the 100 patients experienced relapse, respectively. The 3-year overall survival (OS) and relapse-free survival (RFS) rates were 91.7 % (95 % CI; 83.2 - 96.0) and 69.5 % (95 % CI; 59.0 - 77.9), respectively. In terms of genetic analysis, 21 / 100 (21.0%) patients had KIT exon 17 mutation, 11/100 (11.0%) had KIT exon 8 mutation, 6/100 (6%) had KRAS mutation, 16/100 (16%) had NRAS mutation, 2/100 (2%) had C/EBPA mutation, 2/100 (2%) had WT1 mutation, 6/100 (6%) had ASXL1 mutation, 9/100 (9%) had ASXL2 mutation, and 6/100 (6%) had CSF3R mutation. FLT3-ITD was also identified in 3 of the 100 (3%) patients. In terms of cell surface marker expression pattern, CD19 expression was negative in 59 / 100 (59.0%) patients, and CD56 expression was positive in 43 / 100 (43.0%) patients. Patients with KIT exon 17 mutation were significantly accumulated in the CD19 negative (CD19 (-)) population (18 / 59 vs. 3 / 41, p < 0.001). Survival analysis revealed that KIT exon 17 mutation and CD19 (-) were associated with inferior 3y-RFS (KIT exon 17 mutation: negative vs positive, 74.6 vs 50%, p<0.01, CD19 expression: positive vs negative, 83.1 vs 59.8 %, p<0.01). On the contrary, CD56 (+) was not associated with poor RFS (positive vs negative, 66.1 vs 74.7%, p=0.15). In addition, CD19 (-) was associated with poor RFS even in the patients without KIT exon 17 mutation (positive vs negative, 85.4 vs 65.2%, p=0.04). Finally, CD19 (-) was the sole significant risk factor of relapse (hazard ratio; 3.09, 95% CI; 1.26 - 7.59, p < 0.01) by multivariate analysis. Discussions: This study revealed that CD19 negativity might be a distinct character of poor prognostic subgroup of RUNX1-RUNX1T1-positive AML. On the contrary, CD56 positive was not poor prognostic factor of RUNX1-RUNX1T1-positive AML. Although majority of the patients with KIT exon 17 activating mutation belonged to CD19(-) patients, CD19 (-) patients without KIT exon 17 mutation still showed inferior RFS, suggesting biological difference between CD19 (+) and CD19 (-) RUNX1-RUNX1T1 AML should be investigated. Disclosures No relevant conflicts of interest to declare.
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Klever, Marius-Konstantin, Eric Sträng, Julius Jungnitsch, et al. "Integration of Hi-C and Nanopore Sequencing for Structural Variant Analysis in AML with a Complex Karyotype: (Chromothripsis)²." Blood 136, Supplement 1 (2020): 28. http://dx.doi.org/10.1182/blood-2020-133787.
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Background. Acute myeloid leukemia (AML) with a complex karyotype (CK-AML) is an AML subtype with a still dismal outcome despite recent therapeutic advances. The prognosis is even worse when the underlying structural variants (SVs) lead to an extremely complex pattern of rearrangements, called chromothripsis, with a median overall survival of only 120 days. Except for the presence of inactivating TP53 aberrations in about 70% of all AML-CK cases, the pathogenesis is poorly understood. To gain novel insights into the molecular mechanisms underlying CK-AML reliable high precision SV delineation is needed, which so far has been a major limitation in cancer research. Aim. We developed a SV detection pipeline by integrating Oxford Nanopore Technology (ONT) based whole genome sequencing (WGS) and Hi-C sequencing. This pipeline generated precise characterization of SVs for which the impact on gene expression and the emergence of novel fusion genes was studied by RNA-seq and ONT transcriptome sequencing. Patients and Methods. We applied our WGS and Hi-C SV detection pipeline to a cohort of 11 AML-CK cases. Nanopore DNA Sequencing was performed until a genomic coverage &gt;10x per patient was reached. The samples of 9 patients were also subjected to Nanopore cDNA sequencing for fusion gene analysis and Illumina based RNA-seq for transcript quantification. As controls for Hi-C and Illumina RNA sequencing, CD34+ hematopoietic stem cell enriched samples from five healthy donors were used. Results. Our SV detection pipeline enabled us to fully reconstruct the derivate chromosome structure even of very complex, chromothriptic rearrangements in CK-AML. This enabled us to identify features of chromothripsis, that could previously not be detected using conventual technologies. We found local clustering of breakpoints in three of the patients with up to 31 Inversions and Translocations located in a genomic region of just 2.7 kb. These breakpoints were present in the Hi-C as well as in our Nanopore SV dataset. Our SV pipeline also showed that in these highly clustered regions, the very small rejoined fragments (in many cases less than 1 kb in size) often showed an elevated copy number (CN) state, i.e. small amplifications. We termed this newly discovered phenomenon chromothripsis-in-chromothripsis or (chromothripsis)². The precise knowledge about these breakpoints, which were validated by two different technologies, enabled us to study the pathogenesis of CK-AML at a so far unprecedented resolution. Fusion transcripts could be very precisely mapped and the impact of the breakpoints and CN changes on gene expression levels could be validated, thereby indicating functional relevance of the respective aberrations. Conclusions. The combination of Hi-C and long-read sequencing for SV detection proved to be a powerful tool for precise SV detection. Our SV pipeline allowed us to discover a new level of complexity in chromothripsis. Application of this pipeline to leukemias as well as other types of cancer can improve the precision of SV detection, thereby raising new opportunities for functional interpretation of complex genomic aberrations of pathogenic relevance. Disclosures Döhner: Sunesis Pharmaceuticals: Research Funding; Astex Pharmaceuticals: Consultancy; Pfizer: Research Funding; Bristol-Myers Squibb: Research Funding; Arog: Research Funding; Roche: Consultancy; Novartis: Honoraria, Research Funding; Jazz Pharmaceuticals: Consultancy, Honoraria, Research Funding; Daiichi Sankyo: Honoraria; Abbvie: Consultancy; Agios: Consultancy; Janssen: Consultancy, Honoraria; Amgen: Consultancy, Research Funding; Astellas Pharma: Consultancy; Celgene: Consultancy, Honoraria. Schrezenmeier:Alexion Pharmaceuticals Inc.: Honoraria, Research Funding. Bullinger:Amgen: Membership on an entity's Board of Directors or advisory committees; Astellas: Membership on an entity's Board of Directors or advisory committees; Bristol-Myers Squibb: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees; Seattle Genetics: Membership on an entity's Board of Directors or advisory committees; Janssen: Membership on an entity's Board of Directors or advisory committees; Jazz Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees; Pfizer: Membership on an entity's Board of Directors or advisory committees; Hexal: Membership on an entity's Board of Directors or advisory committees; Gilead: Membership on an entity's Board of Directors or advisory committees; Daiichi Sankyo: Membership on an entity's Board of Directors or advisory committees; Abbvie: Membership on an entity's Board of Directors or advisory committees; Novartis: Membership on an entity's Board of Directors or advisory committees; Sanofi: Membership on an entity's Board of Directors or advisory committees; Menarini: Membership on an entity's Board of Directors or advisory committees.
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Del Principe, Maria Ilaria, Francesco Buccisano, Stefano Soddu, et al. "Pattern of Central Nervous System (CNS) Involvement in Adult Acute Myeloid Leukemia (AML) and Its Impact on Outcome." Blood 128, no. 22 (2016): 2789. http://dx.doi.org/10.1182/blood.v128.22.2789.2789.
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Abstract Background Routine diagnostic lumbar puncture is not recommended in adult patients with acute myeloid leukemia (AML) without Central Nervous System (CNS) symptoms and little is known about the incidence of CNS involvement and its impact on survival in these patients. Furthermore, several studies have demonstrated that flow cytometry (FCM) is superior to conventional cytology (CC) for detection of CNS involvement in lymphoproliferative disorders but the role of this approach for the investigation of cerebrospinal fluid (CSF) in AML is unknown. Design and Methods The aims of our study were 1) to determine the incidence of occult/manifest CNS disease in a homogenous series of AML patients; 2) to correlate CNS disease with clinico-biologic parameters; 3) to examine the impact of CNS involvement on outcome. CSF samples were collected from 98 newly diagnosed AML patients, 62 males and 36 females, median age 53 years (range 18-75). Sixty-five and 33 patients aged <60 and>60 years, respectively.Seventy-one patients received standard and 22 high-dose-ARA-C-based regimens, 5 supportive care. All of 98 CSF samples were examined by CC whereas 90 (91%) also by FCM. CC positivity was defined as unequivocal morphologic evidence of leukemic blast in CSF and/or white blood cells count (WBCc) ≥ 5/µl with less than 10 erythrocytes/µl. A cluster of at least 10 phenotypically abnormal events was regarded as a proof of FCM positivity. Results Sixty-seven patients were CNS negative (CNS-) while thirty-one (31%) were CNS positive (CNS+). Among the last, 10 (10%) were positive on both CC and FCM (manifest CNS+) and 21 (21%) only on FCM (occult CNS+). There was an equal male/female distribution among CNS- and CNS+ patients, as well as median age (52 years, range, 20-71, vs 56 years, range, 18-75, p=NS) and WBCc(27.5 x109/L, range, 1,20-223 x109/L, vs. 11,6 x109/L, range, 0,70-315 x109/L, p= NS) were similar in both groups. Instead, higher levels of lactate dehydrogenase (LDH) were observed among CNS+ than CNS- patients (p=. 01). Forty-seven patients (48%) had monoblastic/monocytic or myelomonocytic AML and belonging to one of these categories was significantly associated with a condition of CNS positivity (55% vs 45%, P = 002). Cytogenetic/genetic data were available in 82/98 (84%). Twenty-for patients (29%), 33 (39%), 12 (14%) and 12 (14%), belonged to the category of favorable, intermediate-I, intermediate-II, and adverse karyotype, respectively. Cytogenetic/genetic characteristics did not differed significantly between CNS+ and CNS- patients. Overall, response rate was 70%, with complete remission rate being not statistically different between CNS+ and CNS- patients (69% vs 81% p= NS). Five-year DFS and OS were found to be significantly shorter in occult or manifest CNS+ patients than in those CNS- (23% vs 50% p= .03 and 19% vs 46%, p=.02, respectively)(Figure 1A and 1B). The prognostic variables achieving a statistical significance in univariate analysis (CNS status, age , WBCc, favorable vs adverse karyotype) were challenged in a multivariate model to determine to what extent they affected treatment outcome. In multivariate analysis, CNS positivity was found to be independently and significantly associated with a shorter duration of DFS.(p=.03 HR= 0.46). Age >50 years was found to be the only independent prognostic factor affecting OS (p=.01 HR= 2.26). Conclusion Our data suggest that incidence of CNS involvement in newly diagnosed AML pts is higher than expected. Regardless of neurologic symptoms, manifest and occult CNS positivity should always be sought at diagnosis since it may affect outcome and influence therapeutic decision. Further prospective studies on larger series are warranted to confirm this data. Figure 1 DFS and OS based on CNS status Figure 1. DFS and OS based on CNS status Disclosures Lo Coco: Teva: Consultancy, Honoraria, Speakers Bureau; Lundbeck: Honoraria, Speakers Bureau; Novartis: Consultancy; Baxalta: Consultancy; Pfizer: Consultancy.
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Cilloni, Daniela, Francesca Arruga, Francesca Messa, et al. "Variable but Consistent Pattern of Meningioma 1 Gene (MN1) Expression in Different Genetic Subsets of Acute Myelogenous Leukaemia and Its Potential Use as Marker for Minimal Residual Disease Detection." Blood 110, no. 11 (2007): 3494. http://dx.doi.org/10.1182/blood.v110.11.3494.3494.
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Abstract Meningioma 1 (MN1) gene overexpression has been reported in acute myeloid leukaemia (AML) patients and identified as a negative prognostic factor. In order to characterize the patients presenting the gene overexpression and to verify if MN1 transcript could be a useful marker for minimal residual disease detection, MN1 has been quantified by RQ-PCR in 136 AML patients of different cytogenetic groups and in 50 normal controls. In 20 patients bearing a fusion gene transcript (FG) suitable for MRD assessment, we performed a simultaneous analysis of the MN1 and of the FG transcript during follow-up. Sequential MN1 and WT1 analysis was also performed in 10 AML patients lacking other molecular markers. The MN1 levels were extremely low in normal samples: the median of 2−ΔΔCt is 4,6 ±2,9 (range 3–10) in PB and 16 ±19,6 (range 6–50) in BM and 12,9 ±4,8 (range 11–19) in normal CD34+ cells. Conversely, about 50% of the AML samples with normal karyotype (NK) showed high expression of the MN1 gene with a median value of 2−ΔΔCt =111±590 (range 52–2352) in BM and 101± 399 (range 12–1136) in PB. All samples carrying the CBFβ-MYH11 FG expressed a significantly higher amount of MN1 transcript as compared to controls (p<0,0001 in both BM and PB): median =1176±1180 (range 362–2272) in BM and 588±401 (range 17–1060) in PB. About 50% of the samples with AML1-ETO FG abnormally expressed MN1: median of 89±58 (range 55–181) in BM and 54±29 (range 21–81) in PB. Finally, the APL samples expressed MN1 values comparable to those of healthy subjects in both BM (p= 0,05) and PB (p=0,08). Interestingly, the paired analysis established a remarkable correlation between MN1 expression in PB and BM with a r value of 0,9627. Stratification of patients according to the presence of FLT3 mutation or ITD demonstrated no significant association between the two abnormalities. In contrast, MN1 overexpression is typically present in patients with mutations in NPM1. 36 out of 47 patients presenting NPM1 mutations were characterized by abnormal expression of MN1. Finally, we were unable to find any significant correlation between EVI-1 and MN1 expression (r= 0,06). To assess the significance of MN1 as a marker for MRD detection in AML, the MN1 transcript was quantified during follow-up of 20 AML patients characterized by the presence of FG (15 CBFβ-MYH11 and 5 AML1-ETO) and 10 patients lacking additional markers monitored by WT1 quantitative assessment, In all cases characterized by FG transcript, the longitudinal pattern of MN1 expression always paralleled that of the FG. Furthermore, MN1 strictly paralleled WT1 in patients without any FG. In all the cases MN1 rose at least two months before relapse. In conclusion, the data obtained show that high levels of MN1 expression are present in 47% of patients with NK primarily in those with wild type NPM1, and in all cases with inv(16). The MN1 levels during follow-up were found to follow the pattern of the other molecular markers (fusion gene transcripts and WT1). Increased MN1 expression in the BM during follow up was always found to be predictive of an impending hematological relapse.
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Biggerstaff, Julie A. Sanford, Weihui Liu, Caron D. Glotzbach, and Lisa G. Shaffer. "Development and Clinical Validation of a “Home-Brew” Dual-Color FISH Probe Assay To Differentiate between ELL and ENL Gene Rearrangements in Leukemia." Blood 106, no. 11 (2005): 3268. http://dx.doi.org/10.1182/blood.v106.11.3268.3268.
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Abstract Patients with t(11;19) leukemias have one of two different translocations resulting in gene rearrangement with the MLL gene on 11q23. The ELL (eleven-nineteen lysine-rich leukemia) gene at 19p13.1, is rearranged in individuals with t(11;19) and acute myeloid leukemia, while the ENL (eleven-nineteen leukemia) gene at 19p13.3, is rearranged most frequently in those patients with t(11;19) and B-cell acute lymphoid leukemia. However, AML and rare T-cell ALL patients have been reported with the ENL gene rearrangement. At the cytogenetic level of resolution, it is not always possible to distinguish which of the two genes is involved in these translocations and precise molecular classification may be necessary for appropriate chemotherapy regimens to be devised and administered. We therefore developed a dual-color FISH (fluorescent in situ hybridization) assay using BACs (bacterial artificial chromosomes) for ELL and ENL. ELL is directly labeled in spectrum orange and ENL is directly labeled in spectrum green and the probes are co-hybridized. The strategy for the assay is to assess the interphase nuclei of patients for gain of an orange signal, indicating an ELL gene rearrangement (3O2G signal pattern), or gain of a green signal, indicating an ENL gene rearrangement (3G2O signal pattern). The gain of signal is due to splitting of the respective locus during the translocation of distal 19p to 11q23. As constructed, the probes showed a 100% specificity for the respective translocations, based on metaphase and interphase analysis of ELL and ENL t(11;19) positive patients; all of the patients were diagnosed with myeloid leukemia. In addition, further comparative validation was performed using the commercially available dual-color breakapart MLL probe (Vysis). Analysis of the ELL positive AML patients with the MLL probe showed the expected abnormal interphase signal pattern (1O1G1Fusion), confirming MLL gene rearrangement. However, the MLL pattern for the single patient with an ENL gene rearrangement and AML showed an alternate interphase pattern, indicating an alternate MLL breakpoint. Analysis of additional patients is required to determine if those with AML and ENL gene rearrangements routinely have a different MLL breakpoint; this difference could potentially contribute to a myeloid disease presentation. In order to establish clinically valid cut-offs for residual disease analysis, interphase validation was performed. Probe specificity was 100%, with a sensitivity of greater than 97% for both loci. This results in a normal cut-off (plus 2 S.D.) of 0.9/200 abnormal cells for ENL gene rearrangement and 2.18/200 abnormal cells for ELL gene rearrangement. In this laboratory’s use, the probe set is slightly more sensitive for disease detection than the commercially available MLL gene probe (Vysis), which has a sensitivity of 96% and a normal cut-off of 1.84/200 cells. This probe set allows quick distinction between ELL and ENL gene rearrangements in patients with acute leukemia.
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Wong, Cynthie, Vincent Funari, and Maher Albitar. "Immunophenotyping Using Targeted RNA NGS Recapitulates Traditional AML and CLL FLOW Fingerprints." Blood 132, Supplement 1 (2018): 5256. http://dx.doi.org/10.1182/blood-2018-99-119085.
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Abstract Introduction Flow cytometry is the gold standard for diagnosing hematologic cancers based on morphologic detection and analysis of a few expensive and delicate immunological markers. On the other hand, targeted RNA sequencing panels are not sample or marker limited; in fact, 50ng of RNA stored for up to 6 months could yield results for thousands of markers. We hypothesized that an RNA-Seq-based targeted immuno-oncology gene expression panel could recapitulate the FLOW diagnostic patterns of AML and CLL routinely used in a clinical laboratory. Methods A custom panel of 2207 genes was constructed including 58 typical FLOW markers and well-referenced immune and oncology markers. Housekeeping genes were added to normalize between batches. A total of 52 CLL, 15 AML, and 20 normal clinical samples were tested in parallel with a clinically validated leukemia/lymphoma flow cytometry panel and targeted RNA-Seq. Paired-end 76 x 76 cycles sequencing was performed using Illumina NextSeq. Bowtie analysis suite was performed to determine gene expression. Unsupervised analysis was first performed to identify patterns associated with clinical diagnosis or sequencing artifacts. Two-way hierarchical clustering of genes having a median expression of >1 fpkm and at least 2 fold differential expression than the median in 10% of the samples revealed a strong CLL profile and a less pervasive AML profile without any supervised analysis. To determine which genes in the profiles were significantly associated with AML or CLL, genes with >5 fold differential expression were assessed after Benjamini-Hochberg correction with single tailed T-tests. Further, each FLOW marker was individually tested using a 1-way ANOVA. Pathway analysis was performed on GO terms using the Fischer exact test. All corrected p-values <0.05 were considered significant. Results In general, the FLOW marker gene expression data highly correlated with protein marker expression and was adequate for rendering proper diagnosis. Overall, CLL had a strong immune-oncology pattern with 10+ flow markers including CD19, CD5, CD2, CD200, CD22, CD79, FCER2, IL3RA, IL2RA, PDCD1, and MS4A1 significantly associated with CLL. In addition, 295 other genes including immune targets like CD74, CD33, CD34, CD48, CD40, and gene targets like PAX5, BCL2, PARP3 further help classify CLL. Forty-three of these genes are involved in immune response pathway (p<1.9x10-17). In contrast, two markers used for FLOW (CD34 and CD52) could classify AML with RNA, and 218 other genes including immune (CD3E/G, CD23, CD48, CD6, CD33) and molecular markers (HOXA10, HOXA9, and TGFBR2) could be used to further classify AML. Interestingly, these genes were significantly enriched for T cell co-stimulation (p<2.0x10-18) and other T cell receptor signaling pathways. To determine whether other immune genes may be used to differentiate CLL or AML, hierarchical clustering of the top 200 genes significantly expressed in either CLL or AML was performed. We could clearly identify two clusters of genes which characterize CLL from other disease types: 1) 110 genes which were highly expressed in CLL, but expressed at low levels in both normal and AML samples, 2) 28 genes with low expression in CLL, but highly expressed in both normal and AML samples. Conversely, few genes were able to characterize AML from normal or CLL samples, including BMP1, NPM2, and FLT3, which were highly expressed in AML samples, but were expressed at low levels in both normal and CLL samples. Conclusion Based on our preliminary study, we have shown that protein marker expression determined by flow are reproduced by our RNA expression panel. Importantly, we are able to classify and diagnose CLL and AML samples based on their RNA expression profiles. Disclosures Wong: NeoGenomics: Employment. Funari:NeoGenomics: Employment.
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Tsaur, Grigory, Olga Plekhanova, Alexander Popov, et al. "Molecular Genetic Characterization of 3'-Deletion of MLL Gene in Infant Acute Leukemia." Blood 120, no. 21 (2012): 2498. http://dx.doi.org/10.1182/blood.v120.21.2498.2498.
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Abstract Abstract 2498 Background. MLL gene rearrangements are associated with unfavorable outcome in infant acute lymphoblastic leukemia (ALL) and have intermediate prognosis in infant acute myeloid leukemia (AML). Application of fluorescence in-situ hybridization (FISH) allows detecting not only conventional MLL rearrangements, but also concurrent 3'-deletion of MLL gene. However, detailed characteristics of infant leukemia carrying 3' MLL deletion remain unclear. Aim. To investigate molecular genetic features of MLL-rearranged infant acute leukemia with concurrent 3' MLL deletion. Methods. 64 patients (27 boys and 37 girls) aged from 1 day to 11 months (median 6.6 months) including 44 ALL patients, 18 AML patients, 1 patient with acute bilineage leukemia and 1 patient with acute undifferentiated leukemia were enrolled in the current study. Chromosome banding analysis was done according to standard procedure. FISH analysis using LSI MLL Dual Color, Break Apart Rearrangement Probe (Abbott Molecular, USA) was performed on at least 200 interphase nuclei and on all available metaphases. Presence of MLL rearrangements was detected by FISH, reverse-transcriptase PCR. In 29 cases long-distance inverse PCR was additionally performed. In case of MLL rearrangement presence standard FISH pattern was defined as simultaneous detection of 3 different fluorescent signals: 1 fused (orange) signal, 1 green signal derived from 3' part of MLL gene, 1 red signal from 5' end of MLL (1F1G1R). MLL rearrangements with concurrent 3' MLL deletion led to 1F1R FISH pattern formation due to lack of green signal. Results. FISH revealed MLL rearrangements in 73% of ALL cases that was higher than frequency of 11q23 translocations detected by conventional cytogenetics — 55%. In MLL-positive cases we found 38 patients (81%) with standard FISH pattern, 7 ones (15%) with concurrent 3'-deletion of MLL gene and 2 (4%) with complex MLL rearrangements. Among patients with 3' MLL deletions there were 1 case with 5' MLL duplication (1F2R) and 1 case with 5' MLL triplication (1F3R). Frequency of 3'-deletions were similar in ALL and AML patients (13% and 15%, respectively). We did not find more than one FISH pattern in bone marrow blast cells of each patient with 3' MLL deletion. In this cohort of patients all blast cells carried concurrent 3'-deletion of MLL gene. Moreover, percentage of blast cells carrying MLL rearrangements did not differ significantly between patients with standard FISH pattern (median 97%, range 22–100%) and 3'-deletion (median 83%, range 13–99%) (p=0.206). 3'-deletion of MLL was not associated with breakpoint position in MLL gene and type of translocation partner gene. MLL translocation partner genes detected in patients with 3' deletions were as follows AF4(n=2), MLLT3(n= 3), MLLT10(n=2). None of the patients with 3'-deletions had reciprocal fusion gene. Initial patients' characteristics (age, sex, WBC count, immunophenotype, CNS-status, type of MLL partner gene) and treatment response parameters (day 8 peripheral blood blast cell count, day 15 bone marrow status, day 36 remission achievement, minimal residual disease status at time point 4) did not differ significantly between 2 groups. Although cumulative incidence of relapse was lower in patients with 3'-deletion as compared to patients with standard FISH pattern (0.31±0.04 and 0.55±0.01, respectively), difference between these two groups was not statistically significant (p=0.359). Conclusion. In our work we characterized rare subgroup of infant MLL-rearranged acute leukemia carrying concurrent 3' MLL deletion. Our data provide additional information of molecular genetic features of acute leukemia in children younger than one year. Disclosures: No relevant conflicts of interest to declare.
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Schoch, Claudia, Mirjam Klaus, Susanne Schnittger, Wolfgang Hiddemann, Wolfgang Kern, and Torsten Haferlach. "Interphase FISH and Comparative Genomic Hybridization Performed in Addition to Chromosome Banding Analysis in AML with Normal Karyotype Detect Prognostically Relevant Chromosome Abnormalities." Blood 104, no. 11 (2004): 2016. http://dx.doi.org/10.1182/blood.v104.11.2016.2016.
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Abstract In AML karyotype abnormalities are not detected in 40 to 45% of cases using classical chromosome banding analysis. For several reasons false negative results might occur in chromosome banding analysis: 1. no proliferation of the aberrant clone in vitro, 2. low resolution due to technical problems or limitations of the method itself, 3. real cryptic rearrangements. In order to determine the proportion of “false negative” karyotypes by chromosome banding analysis we conducted a study using interphase-FISH and comparative genomic hybridization in addition to chromosome banding analysis. In total, chromosome banding analysis have been performed in 3849 AML at diagnosis. Of these 1748 showed a normal karyotype (45.4%). Out of these in 3 cases cytomorphology revealed an APL and in 2 cases an AML M4eo. Using interphase FISH with a PML-RARA or CBFB probe we detected cryptic PML-RARA or CBFB-rearrangements, respectively, in all 5 cases, which were cytogenetically invisible due to submicroscopic insertions. 480 cases of AML with normal karyotype were analyzed for MLL gene rearrangements using FISH with an MLL-probe. 11 cases with a cryptic MLL-rearrangement were detected (FAB-subtypes: M5a: 7, M2: 2, M0: 2). In 273 patients interphase-FISH screening with probes for ETO, ABL, ETV6, RB, P53, AML1 and BCR was performed. In 6 out of 273 (2.2%) pts an abnormality was detectable. In two cases the aberrant clone did not proliferate in vitro: 1 case each with monosomy and trisomy 13. Due to limitations of resolution in chromosome banding analysis translocations or deletions of very small chromosome fragments were only detected with FISH in n=4 cases (ETV6 rearrangements: t(11;12)(q24;p13), t(12;22)(p13;q12), ETV6 deletions: del(12)(p13), n=2). Like interphase-FISH comparative genomic hybridization (CGH) does not rely on proliferating tumor cells but in contrast to interphase-FISH allows the detection of all genomic imbalances and not only of selected genomic regions. Therefore, we selected 48 cases with normal karyotype and low in vitro proliferation (less than 15 analyzable metaphases in chromosome banding analysis). In 8 of 48 cases (16.7%) an aberrant CGH-pattern was identified which was verified using interphase-FISH with suitable probes. In 3 cases a typical pattern of chromosomal gains and losses observed in complex aberrant karyotypes was detected. In one case each a trisomy 4 and 13 was observed, respectively. In one case trisomy 13 was accompanied by gain of material of the long arm of chromosome 11 (11q11 to 11q23). One case each showed loss of chromosome 19 and gain of the long arm of chromosome 10, respectively. In conclusion, CGH in combination with interphase-FISH using probes for the detection of balanced rearrangements is a powerful technique for identifying prognostically relevant karyotype abnormalities in AML assigned to normal karyotype by chromosome banding analysis. Especially this is true in cases with a low yield of metaphases and in AML with a high probability of carrying a specific, cytogenetically cryptic fusion-gene. Thus, in these cases interphase-FISH and CGH should be performed in a diagnostic setting to classify and stratify patients best.
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Sonneck, Karoline, Richard Fritz, Leonhard Muellauer, et al. "Detection of Activated STAT5 in the Cytoplasm of Neoplastic Cells in Patients with AML, CML, and Systemic Mastocytosis." Blood 108, no. 11 (2006): 2305. http://dx.doi.org/10.1182/blood.v108.11.2305.2305.
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Abstract The signal transducer and activator of transcription 5 (STAT5) has recently been implicated as essential pro-oncogenic factor in the pathogenesis of myeloid leukemias in mice (Cancer Cell2005;7:87–99). More recently, STAT5 activation has also been described to occur in human leukemias. However, so far, little is known about the expression of activated/tyrosine phosphorylated STAT5 (pSTAT5) in various myeloid neoplasms and about the distribution of pSTAT5 in the cellular compartments of the normal and leukemic bone marrow (bm). We have examined the expression of pSTAT5 in the bm in patients with acute myeloid leukemia (AML, FAB M0, n=3, M1, n=6, M2, n=4, M3, n=5, M4, n=5, M5, n=4, M6, n=5, M7, n=4), chronic myeloid leukemia (CML, chronic phase, n=4, accelerated phase, n=5, blast phase, n=5), and systemic mastocytosis (SM, n=30), as well as in the normal bm (n=5). Expression of pSTAT5 was determined on paraffin-embedded bm sections by immunohistochemistry using the pSTAT5-specific antibody AX1. In the normal bm, the antibody AX1 was found to react with megakaryocytes and immature myeloid progenitor cells, whereas erythroid cells and mature granulocytic cells did not stain positive for AX1. In patients with AML and CML, the distribution of pSTAT5 showed a similar pattern. In fact, pSTAT5 was found to be expressed in leukemic blast cells without differences among FAB types as well as megakaryocytic cells, but not in erythroid cells. In patients with SM, neoplastic mast cells were found to be immunoreactive for pSTAT5. Interestingly, in all patients and all cells examined, pSTAT5 was found to be localized in the cytoplasm rather than in the nucleus. The cytoplasmic distribution of pSTAT5 in neoplastic cells was confirmed by immunocytochemical staining experiments performed on primary isolated neoplastic cells (AML, CML, mastocytosis) and respective cell lines (U937, KG1, K562, KU812, HMC-1). In each case, the reactivity of neoplastic cells with the AX1 antibody was abrogated by preincubation of the antibody with a pSTAT5-specific blocking peptide. Moreover, the expression of cytoplasmic pSTAT5 in the leukemic cell lines was demonstrable by flow cytometry. To study the molecular mechanisms underlying STAT5-activation in neoplastic cells, Ba/F3 cells with doxycycline-inducible expression of disease-specific oncoproteins, namely BCR/ABL (CML) and KIT-D816V (SM) were employed. Induction of these oncoproteins in Ba/F3 cells resulted in massive activation of pSTAT5 and DNA binding activity as shown by EMSA and supershift assays. In summary, our data show that neoplastic cells in AML, CML, and SM express cytoplasmic pSTAT5, and that disease-related oncoproteins contribute to STAT5-activation. The particular cytoplasmic localization of pSTAT5 in neoplastic cells suggests that apart from its function as a transcription factor, pSTAT5 may have an additional role as a cytoplasmic regulator in these malignancies.
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Varma, Neelam, Chetna Agarwal, Subhash Varma, and Pankaj Malhotra. "How Fast Can Acute Promyelocytic Leukemia Be Diagnosed? - Evaluation of PML Immunofluorescence, Flowcytometric Immunophenotypic Analysis and RT-PCR for PML/RARa." Blood 114, no. 22 (2009): 4674. http://dx.doi.org/10.1182/blood.v114.22.4674.4674.
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Abstract Abstract 4674 Introduction Acute promyelocytic leukemia (APL) constitutes nearly 5-8% of all leukemias, however its frequency is higher in some populations. It is essential to diagnose APL rapidly and accurately as it often presents as a devastating coagulopathy and shows unique sensitivity to all-trans retinoic acid (ATRA). A great deal of morphological, immunophenotypic and cytogenetic heterogeneity of APL adversely affects efforts for providing an accurate and rapid diagnosis. APL is associated with t(15;17)(q22;q12) with generation of a novel PML/RARa fusion protein in 95% cases. Cases positive for t(15;17)/ PML/RARa are ATRA sensitive, while some of those without it [eg t(11;17)(q23:q21)] are not. Several techniques such as karyotyping, fluorescent in situ hybridization (FISH), and reverse –transcriptase polymerase chain reaction (RT-PCR), used for its detection are time consuming, laborious, costly and require specialized laboratories. Lately, immunnostaining methods have been described using PML antibodies for faster diagnosis of APL. The distinction between APL (AML-M3) and non AML-M3 AML is based upon microgranular versus speckled pattern observed in the nuclei of leukemic cells. Only few studies have described use of PML monoclonal antibody (Moab) – PG-M3. Typical flowcytometric (FCM) immunophenotype of APL cells reveals positivity for CD33 and CD13, with negativity for HLA-DR and CD34. RT-PCR (gold standard method) is used to detect classical APL genetic abnormality PML/RARa hybrid transcripts resulting from t(15;17)(q22;q21) and also the alternate translocations associated with APL. We undertook this study to assess the role of PML immunoflourescence (IF), flowcytometry and RT-PCR for quick diagnosis of APL. Patients and methods During last 17 months, peripheral blood and/or bone marrow samples from 93 consecutive acute non-lymphocytic leukemia (ANLL) cases were obtained after informed consent. All the cases were classified using standard morphological criteria (FAB classification). RT-PCR for PML/RARa, FCM immunophenotyping (IP) (Moab panel comprising of CD13, CD33, CD34, HLA-DR, CD56, CD2, CD19, CD14 and CD64) and PML IF (using anti-PML Moab PG-M3 clone) were performed. In the PML IF study, staining pattern of leukemic cells was noted within 2 hours of staining, using Leica DM LB2 epifluorescence microscope equipped with chilled digital color camera and Leica FW 4000 software. In our experience PML-IF could be completed in 2-4 hrs, FCM-IP in 3-5 hrs and RT-PCR in 12-24 hrs. Results 27/93 (29%) cases belonged to AML-M3 category and rest to different categories of AML (M0:1, M1:9, M2: 39, M4:10, M5:6, M6:1). RT–PCR for PML/RARa was positive in 24/27 (88.88%) cases with morphological diagnosis of AML-M3. bcr1 transcripts were detected in 2 (8.33%), bcr2 transcripts in 8 (33.33%) and bcr3 transcripts in 14 (58.33%) among 24 cases. 22/24 cases positive for PML/RARa by RT-PCR showed typical microgranular pattern of nuclear staining on PML IF (91.66% concordance). PML IF did not give any signal in one case and another one showed speckled pattern. In non AML-M3 cases, RT-PCR for PML-RARa was negative in all and PML immunoflourescence staining revealed speckled pattern in 59/66 (89.39%) cases. Typical FCM signature for APL was seen in 86.66% cases positive for PML/RARa by RT-PCR. Two cases were HLA DR positive and one out of these was positive for PML-RARa RT-PCR. Conclusions Immunostaining with PML-antibody was found to be a rapid, simple, cost effective & less time consuming technique to detect PML-RARA with high rate of concordance with the gold standard RT-PCR. This could be routinely applied as an upfront investigation to pick up the APL cases. Disclosures: No relevant conflicts of interest to declare.
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Flaig, Thomas W., Catherine M. Tangen, Maha H. A. Hussain, et al. "Randomization Reveals Unexpected Acute Leukemias in Southwest Oncology Group Prostate Cancer Trial." Journal of Clinical Oncology 26, no. 9 (2008): 1532–36. http://dx.doi.org/10.1200/jco.2007.13.4197.
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Purpose Southwest Oncology Group (SWOG) study 9921 is a randomized, phase III, intergroup study to define the role of adjuvant chemotherapy in patients with high-risk prostate cancer. Patients and Methods We allocated 983 patients with prostate cancer with high-risk features to receive 2 years of androgen-deprivation therapy (ADT) with or without six cycles of mitoxantrone (12 mg/m2) after prostatectomy. Results In January 2007, SWOG 9921 was closed to further accrual after three cases of acute myelogenous leukemia (AML) were reported of a total of 487 patients in the mitoxantrone treatment arm. The key cytogenetic features of these cases included inv(16) in the first case, t(15;17) in the second, and del(5) in the third case. Time from the start of mitoxantrone to the detection of AML was 13, 48, and 72 months, respectively. Before SWOG 9921, there were no cases of mitoxantrone-induced AML reported in patients treated for prostate cancer. Conclusion The emergence of this possible pattern of secondary malignancy emphasizes the importance of randomized controlled trials in defining safety and efficacy of new approaches for patients in the adjuvant setting.
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S, Shukitha. "An Overview on usage of Artificial Intelligence in Banking Industry." Shanlax International Journal of Arts, Science and Humanities 6, S1 (2018): 84–87. https://doi.org/10.5281/zenodo.1403593.
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Banking industry is the backbone of the whole economy. Banking definition reveals the main objective of banking industry that is acceptance of deposits and lending of loans to customers. According to world bank report, in India there are 3.5 ATM’s and 7 branches for 1,00,000 people government aims to include maximum number of people under the banking umbrella and hence, the government has introduced Jandhan Yojana (zero balance account) type of financial inclusion schemes. Nowadays, banks apart from fulfilling their basic objective of acceptance of deposits and lending of loans, they are into providing certain value added services to the customers such as merchant banking, loan syndication, factoring, smart cards, micro finance, leasing, foreign exchange services, portfolio management and fund transferring services. Online banking system has turned as a boon to the banking industry, where latest technologies with Artificial Intelligence have been implemented successfully.”Artificial Intelligence is the theory and development of computer systems able to perform tasks normally requiring human intelligence“. With the introduction of AI in banking sector risk assessment, financial analysis and portfolio management functions are becoming more effective. Chatbots, AML pattern detection techniques, Algorithmic trading, etc. type of technologies are being used in the above areas. This paper wants to reveal the areas where AI has been invested successfully and the future areas where it can be implemented.
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Martelli, Maria Paola, Valentina Pettirossi, Christian Thiede, et al. "Dissecting the Hierarchical Level of Hematopoietic Progenitors' Involvement in AML with NPM1 Gene Mutation and Their Engraftment Potential in Immunocompromised Mice." Blood 114, no. 22 (2009): 480. http://dx.doi.org/10.1182/blood.v114.22.480.480.
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Abstract Abstract 480 Acute myeloid leukemia with mutated NPM1 gene and cytoplasmic nucleophosmin (NPMc+ AML) [Falini B et al, NEJM 2005;352:254-266] is a new entity of WHO classification that shows distinctive biological and clinical features [Falini B et al, Blood 2007;109:874-885] which include negativity for CD34 antigen expression at both immunohistochemistry and gene expression profiling. Flow cytometric analysis shows that, in most NPM1-mutated AML, percentages of CD34+ cells are in the low range (< 5-10%). Detection of NPM1 mutations by molecular techniques and/or immunohistochemistry and Western Blot analysis with specific antibodies provides an important tool for tracking the genetic lesion in leukemic cells at different hierarchical stage. We previously reported involvement by NPM1 gene mutation of the CD34+ cell fraction isolated from patients with NPM1-mutated AML, and, in one case, the involvement, in particular, of the early progenitor CD34+/CD38- [Martelli MP et al, Blood (ASH Annual Meeting Abstracts) 2008;112:307]. Here we expand and confirm our previous observation in 5 cases of CD34-negative NPM1-mutated AML. CD34+/CD38- cells were isolated by either FACS (3 cases, purity >98%) or MACS-sorting (2 cases, purity >92%) and analyzed by molecular analysis and Western Blot with a specific anti-NPM1 mutant antibody, respectively. The presence of either NPM1 gene mutation or mutant protein was demonstrated in all samples analyzed proving the CD34+/CD38- cells belong to the leukemic clone. This cell subpopulation displayed also immunophenotypic features classically associated to leukemic stem cells (LSCs) (CD123+/CD33+/CD90-) in all (16/16) samples analyzed, suggesting they might actually represent the LSCs in NPM1-mutated AML. Indeed, CD34+ cell fraction isolated from NPM1-mutated AML was able to generate leukemia in immunocompromised mice resembling the original patient's disease. However, there is experimental evidence that, at least in some CD34-negative AML, also the CD34- population may contain LSCs. Whether the CD34- cell compartment in NPM1-mutated AML is also able to engraft and outgrow into leukemia in mice remains to be clarified. For this purpose, we assessed the engraftment ability of CD34- cells from 5 NPM1-mutated AML patients. No engraftment was observed in one case. Interestingly, in three patients with myelomonocytic (M4, 2 cases) and myelocytic (M2, 1 case) AML, the CD34- fraction resulted into marrow engraftment by human CD45+/CD33+ myeloid cells that, at morphological and immunohistological grounds, consisted of a mixed population of macrophage cells expressing the CD68 (PG-M1) antigen and mature looking myeloperoxidase (MPO)-positive cells. This pattern possibly reflects short-term engraftment by leukemic cells devoid of self-renewal potential that differentiated into mature elements. However, the neoplastic nature of engrafted cells could be established with certainty only in one case by western blotting detection of NPM1 mutant protein. Immunohistochemistry could not help in these cases to establish the leukemic nature of human cells since terminally differentiated leukemic cells in NPM1-mutated AML show nucleus-restricted NPM1 positivity. In contrast, the pure CD34+ fraction (availabel for comparison in one of these three cases) engrafted as AML with clear blastic morphology and cytoplasmic dislocation of nucleophosmin. In a fourth patient, the highly purified CD34- fraction from relapsed NPM1-mutated AML engrafted in mice with a typical AML picture. These preliminary findings suggest that in general the CD34- fraction from NPM1-mutated AML may have more limited engraftment potential than the CD34+ fraction. Further studies are ongoing to address this issue. Disclosures: Falini: Xenomics: Patents & Royalties.
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Weissinger, Eva M., Meike Hillmann, Hans-Jochem Kolb, et al. "Prospective Evaluation of Graft-Versus-Host Disease-Specific Proteomics Pattern in Patients after Allogeneic Stem Cell Transplantation." Blood 106, no. 11 (2005): 3093. http://dx.doi.org/10.1182/blood.v106.11.3093.3093.
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Abstract We have recently described and published a proteomic pattern specific for the early diagnosis of acute graft versus host disease (aGvHD), based on the application of capillary electrophoresis (CE) and mass spectrometry (MS). Here we report the application of proteome based GvHD-patterns to prospectively collected samples from 86 patients (45 AML, 14 ALL, 8 MM, 5 CLL, 5 SAA, 3 MDS, 3CML, 3 NHL). Fifty three patients were transplanted from matched unrelated donors (MUD), 29 received stem cells from matched related donors (MRD), 3 from haplo-identical donors and 1 was transplanted from a syngeneic sibling. GvHD prophylaxis was methotrexat or mycophenolate and cyclosporin A in the majority of the patients. The follow up is currently between 60 and 549 days after allogeneic HSCT. Urine samples were collected on ice prior conditioning, once a week and monthly after day +60. To avoid degradation of the proteins/peptides the samples were frozen as soon as possible. After thawing and removal of confounding substances, like salts, lipids and of all molecules larger than 30 kDa, the samples were loaded onto the CE, separated according to their charge and, after ionization, directly analyzed in an electrospray ionization time of flight (ESI-TOF) -MS. Between 500 and 2500 peptides and proteins were detected in individual samples. All data generated are stored in a Microsoft MS database. Reproducibility of these analyses is greater 98%. The polypeptide patterns specific for the early detection of acute GvHD were applied to the data generated prospectively and the outcome of these analyses was compared to the clinical diagnosis of aGvHD, sepsis and CMV-reactivation. Forty patients were diagnosed with aGvHD in the current evaluation period, 18 had aGvHD ≥ II. From 461 samples, 68 gave a score with the aGvHD pattern, 15 were false positive, whereas 5 scored false negative. The sensitivity of the aGvHD pattern is more than 90%, the specificity is about 96%. Thus the application of the aGvHD pattern for early recognition of acute GvHD is very useful for predicting the development of aGvHD. In addition, we are currently evaluating samples of patients with chronic GvHD (cGvHD). Seven patients in the prospective cohort have developed cGvHD so far. First results show that in the majority of the patients the cGVHD markers are different from those forming the aGvHD pattern. The polypeptide patterns for aGvHD and cGvHD will be discussed together with those of other complications, like CMV-reactivation or sepsis. Taken together our results demonstrate that the proteome analysis of body fluids collected from patients after HSCT maybe extremely useful for diagnosis of complications.
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Liu, Jinping, Ran Wu, Na Wang, et al. "A Unique Leukemia Mouse Model Established From AML Patient with IDH2 R140Q and FLT3-ITD Mutations Among Other Common AML Mutations." Blood 120, no. 21 (2012): 3579. http://dx.doi.org/10.1182/blood.v120.21.3579.3579.
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Abstract Abstract 3579 Somatic mutations in isocitrate dehydrogenase 1 (IDH1, cytosolic) and IDH2 (mitochondrial homolog) have been associated with acute myeloblast leukemia (AML, 7% and 15% respectively), as well as other malignancies, e.g. gliomas, etc1,2. These mutations lead to 2-hydroxyglutarate (2HG) production in leukemic cells, which can be detected both in leukemic cells and plasma of AML patients. This association has led to speculation that IDH mutations are important in AML leukemogenesis, or oncogenic drivers, and could potentially be used as drug targets for AML treatment. However, these assumptions have never been verified for lack of experimental models. We recently successfully engrafted leukemic cells from bone marrow of an AML patient into immunocompromised NOD/SCID mice. This patient has aggressive disease of poor prognosis, with initial response to standard of care of chemotherapy and subsequent fast recurrence, followed by death. The patient was diagnosed as M5 subtype AML with heterozygous IDH2 R140Q mutation, and also mutations of FLT3-ITD, DNMT3A R882H and NPM1. The transplanted mice (AM7577) developed AML leukemia with typical symptoms (BW loss, hunched, inactivity, labored breathing, ruffled coat and eventual mortality) and with leukemic cells in bone and peripheral organs (e.g. spleen, blood, etc) (gradual detection of leukemic cells in peripheral blood after 30 days). The leukemic cells can serially be passed in mice with 100% take-rate and cause 100% leukemia induced mortality (even with < 1e5 cells), thus creating a renewable and potentially unlimited source of leukemia cells. The leukemic cells in mice are identical to those of the original patient leukemic cells (CD45+, CD33+, CD13+, CD123+, and CD19−, heterozygous IDH2 R140Q mutation, along with FLT3-ITD; DNMT3A R882H and NPM1. This mutation pattern is somewhat in contrast to the previous belief that IDH mutations are mutually exclusive to other common AML mutation3. The engrafted animals produce 2HG in leukemic cells and in mouse plasma, both of which can be readily detected and quantified by high-performance liquid chromatography and mass spectrometry (HPLC/MS). 2HG and cytokines could be useful biomarkers to monitor the disease progression as well as treatment response. The treatment leukemic models in vivo with standard of care (SOC: Ara-C, 5day-on/2day-off dosing scheme) resulted in significant response, manifested by reduction of leukemic load to undetectable in peripheral blood and serum biomarkers for both tested doses (2 and 10mg/kg), as well as extended survivals. Leukemia, however, rapidly relapse after the treatment withdrawal, suggesting that AraC has effect only on the leukemic blasts, but not on leukemic stem cells. The appearance of leukemic blasts in peripheral blood after araC withdrawal can be effectively re-suppressed by araC. Further tests on Flt3 inhibitor for anti-leukemic activities are currently ongoing. In summary, the engrafted mice could serve as a useful experimental model to investigate the role of IDH mutation in leukemogenesis and help to identify new treatment strategies for AML, including compounds targeting IDH mutations and FLT3, etc. This is, to our knowledge, the first reported AML experimental model of featuring IDH mutation. Disclosures: Liu: Crown Bioscience, Inc.: Employment. Wu:Crown Bioscience, Inc.: Employment. Cai:Crown Bioscience: Employment. Wang:Crown Bioscience: Employment. Chen:Crown Bioscience: Employment. Wery:Crown Bioscience: Employment. Chen:Crown Bioscience: Employment. Li:Crown Bioscience, Inc.: Employment.
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Tong, Chunrong, Junfang Yang, Yuehui Lin, et al. "Predictable Recurrence by Regular Monitoring Minimal Residual Disease with Flowcytometry In the Patients with Both AML and ALL: A Single-Center Study of 158 Cases." Blood 116, no. 21 (2010): 1661. http://dx.doi.org/10.1182/blood.v116.21.1661.1661.
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Abstract Abstract 1661 Minimal residual disease (MRD) is the most important factor to predict the prognosis after chemotherapy in the patients with acute leukemia(AL). Among the techniques for MRD detection, flowcytometry (FCM) is both sensitive and feasible tool, which can be used in almost all patients no matter with or without cytogenetic or molecular marker. It has been widely recognized that the MRD by FCM can predict the prognosis of the patients with ALL after several cycles of chemotherapy, but there are few reports on the value of MRD by FCM in AML. To investigate the clinical prognostic value of monitoring MRD regularly with FCM, the correlation of MRD and leukemia-free survival (LFS) in patients with AL without initial high risk factors was studied. From April 2005 to July 2009, 119 newly diagnosed patients (AML 85, ALL 34) and 39 treated cases (AML 19, ALL 20) were included. Those treated patients had attained the first complete remission (CR1) for more than 1 year after chemotherapy in other hospital and then were transferred to our hospital and detected MRD by FCM regularly since then. MRD in bone marrow (BM) was detected every 1 to 3 months in the first year after CR1, and every 2 to 6 months thereafter until hematological or extramedullary relapse or by July 2010. The special antibody combinations were employed for each patient according to aberrant expression of leukemia cells detected by the primary immune phenotype, or antigen drift in the follow-up. MRD was analyzed by the same doctor and assessed according to some abnormal characteristics, including antigen over-expression, antigen low-expression or even loss, co-expression of different lineages or different stage antigens on a single cell, and abnormal patterns in light scatters. MRD+ was defined as the aberrant cells more than 0.01% in BM at the twelfth month for AML and the fifth month for ALL. Among 85 newly diagnosed AML, 33 cases were MRD+ with the median ages 29(4-73)years old, 52 cases were MRD- with the median ages 35(9-68)years old. A total of 2/33 patients in MRD+ group and 40/52 patients in MRD- group remained in LFS. The probability of LFS at 12 months and 24 months was 56% and 16% in MRD+ group, 100% and 84% in MRD- group, respectively (see Figure 1), the difference between MRD+ and MRD- groups was significant statistically (p<0.001). Among 34 newly diagnosed ALL, 13 cases were MRD+ with the median ages 20(12-45)years old, 21 cases were MRD- with the median ages 17(6-35)years old . A total of 1/13 patients in MRD+ and 19/21 patients in MRD- remained in LFS. The probability of LFS at 12 months and 24 months was 46% and 38% in MRD+ group, 95% and 95% in MRD- group(see Figure 2), the difference between MRD+ and MRD- groups was significant statistically (p<0.001). Similar pattern was seen in treated patients. Among 19 treated AML, 0/6 in MRD+ group and 11/13 in MRD- group remained in LFS. Among 20 treated B-ALL, 0/5 in MRD+ group and 14/15 in MRD- group remained LFS. In conclusion, our data indicate that MRD monitored by FCM in both AML and B-ALL correlates to LFS very well. It is important to monitor MRD regularly with FCM in the patients with AL after remission in order to earlier identify high-risk patients for relapse who need intensified treatment, such as allogeneic hematopoietic stem cell transplantation. Disclosures: No relevant conflicts of interest to declare.
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Kaiser, Sabine E., Josef Mautner, Christoph Schmid, Helga Schmetzer, Antonia Gaeta, and Hans-Jochem Kolb. "The delta Assay - A Suitable Tool To Detect the Cellular Immune Response Against Acute Myeloid Leukemia Blasts." Blood 106, no. 11 (2005): 4563. http://dx.doi.org/10.1182/blood.v106.11.4563.4563.
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Abstract Adoptive immunotherapy with donor T-cells has been used successfully for the treatment of both relapsed chronic and acute myeloid leukemia after hematopoietic stem cell transplantation. Here we describe a method for the detection of T cell immunity by the inhibition of cytokine driven growth of blasts. According to the heterogeneous growth pattern of various cases of acute myeloid leukemia (AML) the culture requirements had to be determined individually. The basic cytokine cocktail consisted of SCF (50 ng/mL), GM-CSF (100 ng/mL), IL3 (50 ng/mL), G-CSF (100 ng/mL) and EPO (2 U/mL); this cocktail succeeded to induce blast-proliferation in 15 out of 16 AML samples tested. In a first step we determined optimal growth conditions regarding cell density and the day of linear growth. The peak proliferation was measured by the incorporation of tritium labeled thymidine at various time points. It ranged between days 2 and 5. In a second step, donor cells were stimulated for 10 days with irradiated AML-cell lines (MonoMac6, THP1) or patient derived AML blasts. After harvesting, the donor cells were irradiated (15 Gy) and co-cultured with the AML-cell lines, or the patients cytokine stimulated blast suspensions. Inhibition of growth was measured by the incorporation of tritium labeled thymidine at the previously determined day of linear proliferation of the AML-blasts. A significant inhibition of blast proliferation was observed in the co-culture of normal T cells with both AML-cell lines. In HLA-identical combinations T cells were primed by dendritic cells derived from AML blasts and re-stimulated on days 6 and 10. T cells primed by this method inhibited the growth of AML blasts in higher effector target ratios, in lower ratios no inhibition or even stimulation was observed. In summary, the delta-assay is a suitable tool to monitor specific anti-leukemic immune responses of donor lymphocytes even against very heterogeneous blast populations, if individualized culture conditions are considered.
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Cho, Young-Uk, Hyun-Sook Chi, Sang Hyuk Park, et al. "Comparative Analysis Of Cytogenetic Evolution Patterns During Relapse In The Hematopoietic Stem Cell Transplantation and Chemotherapy Settings Of Patients With Acute Leukemia." Blood 122, no. 21 (2013): 1320. http://dx.doi.org/10.1182/blood.v122.21.1320.1320.
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Abstract Background Despite significant therapeutic progress, many patients with acute leukemia die from deteriorating disease after relapse. Relapse of acute leukemia has repeatedly been associated with cytogenetic clonal evolution. However, only a few studies focused on the direct comparison of cytogenetic evolution patterns during relapse of leukemia after hematopoietic stem cell transplantation (HSCT) and conventional chemotherapy. Thus, we performed comparisons of the cytogenetic patterns of patients with acute leukemia both at diagnosis and relapse after HSCT or standard chemotherapy (SC). Cytogenetic patterns with FLT3 mutational instability were also compared. Methods This retrospective analysis was based on a total of 516 patients. Among them, 349 were diagnosed with acute myeloid leukemia (AML) who developed relapse after HSCT (n = 125) or SC (n = 224); 167 were diagnosed with acute lymphoblastic leukemia (ALL) and developed relapse after HSCT (n = 63) or SC (n = 104). Cytogenetic analysis and FLT3 mutation detection were performed according to standard methods. Results In patients with AML as shown in Table 1, differences in the karyotypes between diagnosis and relapse were more frequent in the HSCT cohort than in the SC cohort (53.2% vs. 40.1%, respectively; P = 0.035). Development of more than three new cytogenetic changes was also more frequent in the HSCT cohort than in the SC cohort (31.4% vs. 15.3%, respectively; P = 0.046). Overall, FLT3 mutation instability did not correlate to the clonal cytogenetic changes. However, restricted to patients showing FLT3 mutation instability, tyrosine kinase domain (TKD) mutation was more frequently involved in patients with cytogenetic changes compared to those without (46.2% vs. 5.0%, respectively; P = 0.008) (Table 2). Conversely, in patients with ALL, frequency and pattern of the cytogenetic alterations between diagnosis and relapse did not differ between HSCT and SC cohorts. For both types of acute leukemia, the median number of cytogenetic alterations increased from 1.0 at diagnosis to 2.0 at relapse (P < 0.001) in the HSCT cohort. The increasing number of cytogenetic alterations was also seen in the SC cohort (P = 0.011). Conclusions Higher frequencies of clonal cytogenetic changes and more complex cytogenetic patterns were observed in the HSCT cohort compared to the SC cohort. This finding may be associated with an adverse outcome for relapse patients after HSCT. FLT3 TKD mutation could have a certain role in promoting cytogenetic progression in a subset of AML patients. In addition, our data extends the findings of recent previous studies, which described patterns of clonal evolution in acute leukemia patients. Disclosures: No relevant conflicts of interest to declare.
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Zini, Gina, Elena Rossi, Mariagrazia Garzia, et al. "Peculiar Morphological, Cytochemical, Biochemical and Immunophenotyping Features in One Case of Acute Myeloid Leukaemia at the Onset." Blood 106, no. 11 (2005): 4479. http://dx.doi.org/10.1182/blood.v106.11.4479.4479.
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Abstract On July 28th 2005, a 53-year old woman was admitted to our Hematology Dpt. with fever (37.5 C), fatigue, marked gingiva hypertrophy, mild hepatomegaly and absence of lymphoadenopaty and splenomegaly. Laboratory tests showed high leukocyte count of 182 x109/L, moderately increased platelet count (438 x109/L), normocytic anemia (10,9 g/dl, MCV 84 fl). Biochemical parameters, including lysozyme, were in the normal range except for high level of lactate dehydrogenase (2524 UI/l). Peripheral blood (PB) smear showed the presence of 94% of blasts, without evident granules. At the cytochemical myeloperoxidase staining blast cells revealed the presence of single or multiple Auer rods, while cytoplasmic granules were positive to the α-naphtyl butyrate esterase. In the bone marrow (BM) aspirate blasts were 80% showing same morphological and cytochemical features as in peripheral blood, while the granulocytic maturating series was 14%. No evidence of morphologically identifiable monocytic series in PB neither in BM. Megakariocytes were slightly increased and there was trilineage dysplasia. Blast immunophenotyping in PB and in BM was as follow: CD45, CD33, CD13, CD7 and HLA-Dr positive, 36% MPO positive and 48% co-express CD11c an CD117. Moreover blasts were CD14, CD19 and CD56 negative. FAB diagnosis was AML-M4. Interphase Fluorescence in situ Hybridization (FISH) for detection of AML1/ETO and bcr/abl gene rearrangements was negative. After treatment with 2 g hydroxyurea and leucapheresis, the WBC count was still high (140 x109/L) while after 1g of aracytin, WBC count decreased to 31000/mm3. Patient is presently enrolled into the GIMEMA protocol AML-12. This case shows at the onset following abnormalities: - an increased platelet count is a rare evenience in an AML at the onset; - negativity for lysozyme; - absence of CD14; - presence of Auer rods in quite all the blast population, which is a very rare evenience associated with monocytic subtypes of acute myeloblastic leukemia (AML); - atypical homogenous cytochemical pattern of blasts, showing Auer rods MPO positive and granules ANBE positive; - absence of any morphological differentiation of monocytic series. S4479
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Mishra, Shruti, Kishore Kumar, Ashutosh Panigrahi, Prabodh Das, Somanath Padhi, and Gaurav Chhabra. "The Utility of Leucocyte Cell Population Data and Scattergram in Rapid Identification of Acute Promyelocytic Leukemia." Blood 136, Supplement 1 (2020): 19–20. http://dx.doi.org/10.1182/blood-2020-142498.
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Introduction: Acute promyelocytic leukemia (APL) is one of the hematological emergencies requiring early detection and treatment. The hematology laboratory plays a pivotal role in management. The correct morphological identification of APL is made even before the confirmation by the cytogenetic or molecular analysis. Automated hematology analyzers are the backbone of the hematology laboratories. Other than providing routine counts and differentials, they are also capable of producing sufficient information in the form of leucocyte cell population data (CPD) scatter plots depicting the leucocyte subpopulation. These data are primarily underutilized, particularly in assessing the lineage of various hematopoietic neoplasm. Here we analyzed the utility of leucocyte CPD and the pattern of scatter plot in Sysmex XN 1000 analyzer for the detection of acute promyelocytic leukemia. Materials and Results: We included 100 controls and 100 AML cases. Leucocyte CPD parameters were compared along with scatter plots from the WDF channel from Sysmex XN1000 analyzer were recorded and analyzed. Immunophenotyping and molecular analysis confirmed the diagnosis in all but two cases, where the patients expired within a few hours of admission in the emergency. Out of a total of 100 cases, 22 were labeled as APL, 63 as AML, and 15 as AML M4/M5. Among the APL cases, in 19 cases, immune-phenotyping was done on peripheral blood, and the remaining three bone marrow aspirate samples were used. The scatter-plot analysis of APL cases showed a characteristic pattern which differentiated this entity from other acute myeloid leukemias. In the case of APL, on Side Fluorescence Light Scatter (SFL) Vs. Side scatter (SSC), the abnormal promyelocytes occupy the space above neutrophils and to monocytes' right, where the immature precursors of myeloid are usually found. In cases where the total leukocyte count was within the normal range, a hiatus between these abnormal promyelocytes and neutrophils are seen. This finding was consistent in all cases irrespective of the morphological variation. On the other hand, other variants of acute myeloid leukemias along with monoblastic leukemias, the scatter plots show the blasts in the monocytic region with extension to the ceiling. On comparison, the myeloblast and monoblasts lie more towards the y-axis than abnormal promyelocytes. All the CPD parameters were compared against the control group and the AML group. The cell population data showed a significant difference in all the parameters representing the side fluorescence, size and granulations except LY-WX (width of dispersion of side scatter) when compared to normal healthy controls. Amidst all the AML cases, APL significantly differed from other cases in NE-SFL, LY-WY, and MO-WZ. AML M4/M5 and APL also varied in LY-Y, NE-WY, NE-WZ, and MO-WY. AMLs excluding AML M4/M5 showed additional variation in MO-Y and MO-Z. Discussion: APL cases have a characteristic scatter plot patter on SFL Vs. SSC. The scatter-plot shows a tear-drop like a collection of cells. These cells have high side scatter and high lateral fluorescence owing to the large size of the cells, apple-core like the nucleus, and dense granulations. The differences in the CPD parameters among APL cases and controls and other acute myeloid leukemia can be attributed to these morphological features of the abnormal promyelocytes. Haider et al. found a significant difference between NE-SFL and other myeloid leukemia. [1] This was in corroboration with our findings. Another research done by Park et al. showed that NE-SFL and MO-WX had the highest sensitivity and specificity in differentiating APL from other AML. [2] Conclusion: We conclude that the new age automated cell counters provide numerous data that remains unexplored and can be utilized further along with the information from the scatter plots to make a rapid diagnosis of Acute Promyelocytic Leukemias. References: Haider R.Z., Ujjan I.U., Shamsi T.S. Cell Population Data - Driven Acute Promyelocytic Leukemia Flagging Through Artificial Neural Network Predictive Modeling. Translational Oncology, 2020;13(1):11-16. Park SH, et al. Cell population data NE-SFL and MO-WX from Sysmex XN-3000 can provide additional information for exclusion of Acute Promyelocytic Leukemia from other Acute Myeloid Leukemias: A Preliminary Study. Ann Lab Med 2016;36:607-610. Figure Disclosures No relevant conflicts of interest to declare.
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Man, Tsz-Kwong, Mohammad Javad Najaf Panah, Jessica L. Elswood, Pavel Sumazin, and Michele S. Redell. "Cite-Seq Reveals Distinct Patterns and Potential Mechanisms of Relapse in Pediatric AML." Blood 138, Supplement 1 (2021): 3458. http://dx.doi.org/10.1182/blood-2021-152646.
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Abstract Introduction - Acute myeloid leukemia (AML) is an aggressive disease with a relapse rate of approximately 40% in children. Progress in improving cure rates has been slow, in part because AML is very heterogeneous. Molecular studies consistently show that most cases are comprised of distinct subclones that diminish or expand over the course of therapy. Single-cell profiling methods now allow parsing of the leukemic population into subsets based on gene and/or protein expression patterns. We hypothesized that comparing the features of the subsets that are dominant at relapse with those that are dominant at diagnosis would reveal mechanisms of treatment failure. Methods - We profiled diagnosis-relapse pairs from 6 pediatric AML patients by Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-Seq). All patients were treated at Texas Children's Cancer Center and consented to banking of tissue for research. CITE-Seq was performed by Immunai (New York, NY) using a customized panel of 65 oligonucleotide-tagged antibodies, the 10x Genomics Chromium system for single-cell RNA library generation, and the Novaseq 6000 for sequencing. After data cleanup and normalization, clustering by scRNA-seq was done using the Seurat package. Cell-type identification of clusters was facilitated by published healthy bone marrow scRNA-seq datasets (van Galen et al, Cell 2019). Differentially expressed genes (DEGs) and proteins (DEPs) between diagnosis and relapse were determined using Wilcoxin ranked sum tests. Results - We generated single-cell transcriptomes for a total of 28,486 cells from 12 samples, with a mean of 2373 cells and 1416 genes per sample. Samples were integrated with batch effect correction, producing 30 distinct clusters (cell types) in total (Figure 1A). Cell types with expression profiles consistent with lymphocytes and erythroid precursors were identified in multiple patients, whereas AML cell types tended to be specific to individual patients (Figure 1B). For patients TCH1, TCH2 and TCH3, the most abundant cell types at diagnosis were rare at relapse, and cell types that were rare at diagnosis became dominant at relapse. For these 3 cases, we identified DEGs between the dominant diagnosis cell types and dominant relapse cell types. We found 18 genes that were upregulated at relapse in at least 2 of the cases. Several genes related to actin polymerization were enriched (ARPC1B, ACTB, PFN1), possibly reflecting an enhanced capacity for adhesion and migration. Also of note, macrophage migration inhibitory factor (MIF) and its receptor CD74 were upregulated at relapse, suggesting a role in chemoresistance. For patients TCH4, TCH5 and TCH6, the same cell types that were abundant at diagnosis were also abundant at relapse, and few genes were significantly altered between diagnosis and relapse in multiple cases. Only SRGN, which encodes the proteoglycan serglycin, and GAPDH were altered in 2 of these 3 cases, and both were downregulated at relapse. We performed similar comparisons to identify proteins that were differentially expressed between diagnosis and relapse pairs. The number of DEPs between the dominant diagnosis and relapse cell types ranged from 0 (TCH1 and TCH6) to 5 (TCH2). The only protein altered in more than one case was CD7, which was enriched at relapse in TCH2, TCH3 and TCH4. Conclusions - From CITE-Seq profiling of 6 pediatric AML cases we identified two distinct patterns of relapse. For 3 cases, relapse occurred by expansion of a subset that was small but present at diagnosis. Enrichment of genes associated with adhesion and survival signaling suggests that these cells survived because they were well-equipped to take advantage of interactions with the microenvironment. For 3 other cases, the population that was dominant at diagnosis persisted and expanded at relapse with few substantial changes in gene or protein expression profiles. This pattern suggests that these AML cells were a priori equipped to survive chemotherapy, even though bulk disease levels were transiently reduced below the limit of detection. Most profiled proteins did not change substantially between diagnosis and relapse. An exception is CD7, which was enriched at relapse in 50% of our cases and represents a potential therapeutic target. Analysis of more cases will refine these relapse patterns, reveal potential mechanisms of chemoresistance and inform the development of novel therapies. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.
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Fitzgibbon, Jude, Matthew Smith, Rachael Arch, et al. "Development of a Human Acute Myeloid Leukaemia Screening Panel and Identification of Novel Gene Mutations." Blood 104, no. 11 (2004): 2991. http://dx.doi.org/10.1182/blood.v104.11.2991.2991.
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Abstract Mutation studies in Acute Myeloid Leukaemia (AML) are complicated by the existence of distinct morphological and cytogenetic subtypes; consequently although mutations in any one gene may occur in only 5% of AML, the frequency of mutation may differ both between the different FAB-types and cytogenetic risk groups studied. It is important therefore not only to validate the screening methodology used but also the suitability of the patient panel tested. A screening panel was developed allowing detection of novel recurring gene mutations within samples derived from patients with AML. Mutation analysis of 6 previously described genes (RUNX1, FLT3, KIT, CEBPA, PTPN11, NRAS) and 2 candidate genes (CCND3, FES) were carried out in a cohort of 175 AML samples representing all FAB types (except M3) and cytogenetic risk groups using a combination of SSCP, DHPLC and sequence analysis. One hundred and fifteen mutations were identified in 97 (55%) patients comprising 81 patients (46%) with one mutation, 14 patients (8%) with 2 mutations, and 2 patients (1%) with 3 mutations. Fifty-five out of 88 (63%) patients with normal karyotype AML had at least one mutation. There was was a weak negative association between FLT3 ITD and loop mutation (p = 0.095), a positive association between KIT mutation and favourable risk cytogenetics (p = 0.001), CEBPA mutation and intermediate risk/normal cytogenetics (p = 0.045) and PTPN11 mutation and poor risk disease (p = 0.001). The frequency of individual gene mutation was in accordance with previously published studies. Three novel mutations of FLT3 (Y589D, D839G, Y842H) were detected in 4 patients that would have been overlooked by conventional gel electrophoresis techniques. A single in frame 51bp deletion of nucleotide 939 – 990, resulting in a deletion of 17 amino acids at the carboxyl-terminus of the cyclin D3 protein was identified in a single patient. Overall, both the pattern and mutation frequencies reported in this cohort are similar to those in the literature supporting its further use as an investigational tool in the evaluation of candidate genes in the genesis of myeloid malignancy.
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Fogelstrand, Linda, Sara Ståhlman, Tore Samuelsson, Jonas Abrahamsson, and Lars Palmqvist. "Identification Of Leukemia-Specific Mutations For Detection Of Minimal Residual Disease In Acute Myeloid Leukemia Using Cell Sorting and Whole Exome Sequencing." Blood 122, no. 21 (2013): 2575. http://dx.doi.org/10.1182/blood.v122.21.2575.2575.
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Abstract The introduction of next generation sequencing techniques into the field of leukemia research has revealed that acute myeloid leukemia (AML) is characterized by a limited number of somatic mutations, in most cases single nucleotide variations (SNVs). In addition to providing insight into the pathogenesis of AML, this information can potentially be used for detection of small amounts of leukemic cells in follow-up samples (minimal residual disease, MRD). The aim of this study was to identify leukemia-specific mutations in AML cells that can serve as leukemia-specific MRD-markers. Identification of leukemia-specific mutations was performed using whole exome sequencing of DNA from sorted leukemic cells and comparison with sorted lymphocytes from the same individual. Cells were obtained from 8 cases of AML, age 30-71 years old, from blood samples taken at the time of diagnosis of AML. Cell sorting was carried out by fluorescence activated cell sorter (FACS), where leukemic cells were defined by their FSC and SSC properties and expression of CD45, CD34, CD117, and HLA-DR. Lymphocytes were sorted based on FSC, SSC and CD45 expression. Purity of cell populations were >98% for leukemic cells and >99% for lymphocytes (with undetectable amounts of leukemic cells). Exome sequencing of sorted cell populations was performed on the Illumina platform with HiScanSQ yielding around 4^107reads per sample. Data were quality assessed by FastQC, aligned to the reference human genome, processed for PCR duplicate removal, variant calling with Genome Analysis Toolkit (GATK) package, annotation of variants with ANNOVAR, and verification in Integrative Genomic Viewer. SNVs and short insertions or deletions present in the dbSNP database were excluded and the resulting SNVs and short insertions and deletions with minimum coverage of 10 were compared between leukemic cells and lymphocytes from the same individual. Leukemia-specific heterozygous mutations were defined as present in >40% of the reads in the leukemic cell sample and present in none of the reads from the corresponding lymphocyte sample. By using these rather strict criteria at least three leukemia-specific SNVs were found in each AML case. Leukemia-specific SNVs (with coverage spanning between 10 and 250) were detected in recurrently mutated genes but also in genes not previously reported to be mutated in AML, e.g. CNNM4, GLYAT, NCKAP1L, PPBP, and PRB1. In the case of previously reported recurrently mutated genes in AML, at least one SNV was found in most AML cases. SNVs in recurrently mutated genes were found to be leukemia-specific in most cases, but in some cases, including PRPF40B, ETV6, and EZH2, SNVs were present in a heterozygous pattern in both leukemic cells and in lymphocytes, indicating that they are germ-line mutations. Genes with leukemia-specific insertions or deletions included NPM1, STAG2, RUNX1, and BCOR. The finding of the insertion in NPM1 in two cases was confirmed by detection of the insertion with conventional fragment analysis used in our clinical laboratory. When the same data analysis was used on exome sequencing data of neutrophilic cells and lymphocytes sorted from normal control samples (n=2), no SNVs or short insertions or deletions were found to differ between these two cell populations. Our results show that by using exome sequencing on sorted cell populations with high purity, leukemia-specific mutations can be identified in AML samples already at diagnosis without the need for additional sampling of normal material or access to remission samples. Information on leukemia-specific mutations at diagnosis could provide a basis for detection of MRD in follow-up samples, either by polymerase chain reaction or targeted deep sequencing. Disclosures: No relevant conflicts of interest to declare.
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Figueroa, Maria E., Tamer Fandy, Melanie J. McConnell, et al. "Myelodysplastic Syndrome (MDS) Displays Profound and Functionally Significant Epigenetic Deregulation Compared to Acute Myeloid Leukemia (AML) and Normal Bone Marrow Cells." Blood 110, no. 11 (2007): 345. http://dx.doi.org/10.1182/blood.v110.11.345.345.
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Abstract MDS are a clinically heterogeneous group of clonal disorders, which share a high frequency of progression to secondary AML (MDS-AML). In contrast to de novo AML, MDS and MDS-AML are uniformly resistant to conventional chemotherapy. Among the few active drugs in MDS are the nucleoside analog DNA methyltransferase inhibitors (MTIs) 5-azacytidine and decitabine. Since tumor suppressor genes such as CDKN2B can be silenced by DNA methylation in MDS, it is believed that reversal of DNA methylation by MTIs might contribute to their anti-tumor effects. However, it is not clear whether methylation-dependent silencing of specific genes are predictive of response or even correlate with response to MTIs. Like MDS, AML also presents epigenetic silencing of CDKN2B and other genes; yet appear to not be as sensitive to MTIs. Given the particular sensitivity of MDS to MTIs and its resistance to standard AML chemotherapy, we hypothesized that MDS is a biologically distinct disease from AML due largely to extensive epigenetic deregulation, which is missed by single locus studies. In order to test this we studied DNA methylation levels at 24,000 gene promoters in 13 MDS pts. and 16 de novo normal karyotype AML cases, and compared and contrasted these to CD34+ bone marrow cells from 8 healthy donors. For this we used the HELP (HpaII tiny fragment Enrichment by Ligation-mediated PCR) assay, a robust method for detection of whole-genome DNA methylation, and using MassArray quantitative methylation for single locus validation. Remarkably, MDS was found to have a far greater number of hypermethylated genes than AML or normal CD34+ cells, while AML and CD34+ cells had similar number of methylated genes (MDS vs. AML: 6303 vs. 4177 promoters, p=0.026; MDS vs. normal CD34+ cells: 6303 vs. 4296 promoters, p=0.056). Using a moderated T test, an aberrant methylation signature of 736 genes (p<0.0000001) was identified in MDS vs. normal CD34+ cells, reflecting extensive epigenetic deregulation in this disease, including p16, CEBPZ, MSH2, CHES1, AKT1, Caspase2, BMP3, DAP and MYOD1. A comparison between MDS and de novo AML identified 498 genes (p<0.00005) differentially methylated, with a clear predominance of hypermethylated promoters in MDS vs. de novo AML. These genes included RUNX2 and 3, GFI1, DAPK2, MDM2, TGFA, CEBPZ and SHARP. Finally, a comparison between normal CD34+ cells and de novo AML demonstrated an aberrant pattern of methylation in 341 genes (p<0.00001), including CXCL1 and 5, PPARD, Caspase 2 HOXA4 and HOXA10, GFI1 and the MLL translocation partner Septin11. 7 of the 13 MDS cases were also examined for gene expression using the Affymetrix Hgu133plus2 array. A significant proportion of the genes that had been found to be methylated were also found to be underexpressed, including p16, DAP, BMP3, HOXA2 and HOXB8. Taken together, our data show that MDS is a unique and distinct biological entity than de novo AML featuring profound and functionally significant genome wide epigenetic deregulation. While the de novo AML methylation profile was clearly different from normal CD34+ cells, it was not as severely altered as MDS. These data also suggest that MTIs are most likely uniquely active in MDS due to their DNA methyltransferase activity.
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Sun, Yang, and James R. Cook. "Comparison of Fluorescence In Situ Hybridization for EGR1 Vs CSF1R for the Detection of Del(5q) in Myelodysplasia and Acute Myeloid Leukemia." Blood 112, no. 11 (2008): 1641. http://dx.doi.org/10.1182/blood.v112.11.1641.1641.
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Abstract The detection of del(5q) in myelodysplastic syndrome (MDS) provides useful information to guide the choice of therapy, given the efficacy of lenalidomide in cases containing this abnormality. Fluorescence in situ hybridization (FISH) analysis offers the opportunity to specifically detect chromosomal abnormalities much more rapidly than metaphase cytogenetics, which may have a turnaround time of several weeks. However, it is currently unclear which chromosomal loci are the most appropriate to examine for detection of del(5q) in routine practice. The breakpoints on chromosome 5q are heterogeneous and two commonly deleted regions (CDR) have been described. The first CDR occurs in acute myeloid leukemia (AML) and high grade MDS and encompasses a region at 5q31 including the EGR1 locus. A second CDR, occurring in at least some cases reported as 5q- syndrome, centers around 5q33 and includes the CSF1R locus. We therefore examined whether FISH studies for EGR1, CSF1R, or a combination of both probes would provide the greatest clinical utility for detection of del(5q). 51 cases of myeloid neoplasms with del(5q) by metaphase cytogenetics were analyzed, including 5q- syndrome (n=8), refractory anemia with excess blasts (RAEB, n=8), refractory cytopenia with multilineage dysplasia (RCMD, n=6), MDS unclassifiable (n=1), therapy related MDS (n=1), myelodysplastic/myeloproliferative overlap syndromes (MDS/MPD, n=6), and AML (n=21). FISH studies using EGR1/D5S23, D5S721 and CSF1R/D5S23, D5S721 probes (Abbot Molecular, Abbot Park, IL) were performed on archival bone marrows (45 coverslip aspirate smears, 4 cytogenetic culture cell pellets, and 2 formalin fixed paraffin embedded clot sections). Normal ranges were established for each probe by analysis of appropriate negative control samples. Deletion of the EGR1 locus was detected in 49/51 (96%) cases, including each case of 5q- syndrome. The CSF1R locus, which could be analyzed in 48 cases, was deleted in 44 cases (92%). In cases with concordant results, a similar percentage of abnormal nuclei was identified with each probe. Two cases (1 MDS/MPD and 1 AML) displayed deletion of the EGR1 locus but a normal pattern for CSF1R. Two cases (1 AML and 1 MDS/MPD) showed no evidence of EGR1 or CSF1R deletion despite a del(5q) identified by metaphase cytogenetics. In conclusion, FISH for EGR1 is sufficient to successfully detect del(5q) in the vast majority of cases of MDS and AML containing this abnormality, including at least most cases of 5q- syndrome. Additional FISH studies for the CSF1R locus did not increase the diagnostic yield. Further studies will be required to determine if deletions of 5q involving EGR1 but not CSF1R influence the response to lenalidomide. A small number of cases of del(5q) are detected only by metaphase cytogenetics, possibly due to a small number of abnormal cells present prior to in vitro culture.