Acute Myeloid Leukemia M2 Research Articles

MiRNA-142-3P and FUS can be Sponged by Long Noncoding RNA DUBR to Promote Cell Proliferation in Acute Myeloid Leukemia.

Acute myeloid leukemia (AML) represents a frequently occurring adulthood acute leukemia (AL). Great progresses have been achieved in the treatment of AML, but its pathogenic mechanism remains unclear. This study reported the biological functions of lncRNA DUBR in AML pathogenic mechanism. As a result, lncRNA DUBR showed high expression level within AML, resulting in poor prognosis, especially in M4 AML. In vitro studies elucidated that knockdown of DUBR with small interfering RNA (siRNA) resulted in the suppression of survival and colony formation ability, as well as induction of apoptosis, in AML cells. RNA pull-down assay and computational revealed that DUBR could sponge with miRNA-142-3P and interact with FUS protein. MiRNA-142-3P have a negative correlation with DUBR and overexpression of miRNA-142-3P inhibited cell growth in AML. Meanwhile, DUBR promoted the expression of FUS protein, targeting inhibition of FUS significantly promoted cell apoptosis in AML cell lines. In conclusion, these results revealed new mechanism of lncRNA DUBR in AML malignant behavior, and suggested that the manipulation of DUBR expression could serve as a potential strategy in AML therapy.

Prevalence and Clinical Risk Factors for Invasive Aspergillosis in Patients with Acute Leukemia at Srinagarind Hospital, Khon Kaen University

Objective: Invasive aspergillosis (IA) is the most common fungal infection in patients with acute leukemia and associated with high mortality rate. We aimed to report the prevalence and clinical risk factors of IA in patients with acute leukemia at Srinagarind Hospital, Khon Kaen University. Materials and Methods: A retrospectively study was performed in adult acute leukemia patients who were diagnosed with IA from January 2006 to December 2015 at Srinagarind Hospital, Khon Kaen University. IA was defined by the European Organization for Research and Treatment of Cancer and the Mycoses Study Group. Clinical characteristics and laboratory data were analyzed. Multivariate logistic regression analysis was used to determine the associations. Results: A total of 355 patients with acute leukemia, invasive aspergillosis was found in 39 patients (11%). The median age was 46 years (IQR of 33 to 59 years). Gender distribution was equal between males and females. Most common type of leukemias in the study are acute myeloid leukemia non-M3 (50.7%), followed by acute myeloid leukemia M3 (24.8%), and acute lymphoblastic leukemia (24.8%) respectively. Using logistic regression analysis, patients who had neutropenia at the time of IA diagnosed (adjusted OR 3.54; p=0.014), hepatitis B viral infection (adjusted OR 3.13; p=0.03), and working as construction worker (adjusted OR 23.08; p<0.001) remained the risk factor of IA. Conclusion: The prevalence of IA in Thai patients with acute leukemia is modest. Neutropenia, hepatitis B viral infection and construction worker were significant clinical risk factors for IA in patients with acute leukemia. Keywords: Invasive aspergillosis; Acute leukemia; Srinagarind Hospital

Venetoclax and azacitidine compared with induction chemotherapy for newly diagnosed patients with acute myeloid leukemia

AbstractVenetoclax (ven) plus azacitidine (aza) is the standard of care for patients with newly diagnosed acute myeloid leukemia (AML) who are not candidates for intensive chemotherapy (IC). Some patients who are IC candidates instead receive ven/aza. We retrospectively analyzed patients with newly diagnosed AML who received ven/aza (n = 143) or IC (n = 149) to compare outcomes, seek variables that could predict response to 1 therapy or the other, and ascertain whether treatment recommendations could be refined. The response rates were 76.9% for ven/aza and 70.5% for IC. The median overall survival (OS) was 884 days for IC compared with 483 days for ven/aza (P = .0020). A propensity-matched cohort was used to compare outcomes in the setting of equivalent baseline variables, and when matched for age, biological risk, and transplantation, the median OS was 705 days for IC compared with not reached for ven/aza (P = .0667). Variables that favored response to ven/aza over IC included older age, secondary AML, and RUNX1 mutations. AML M5 favored response to IC over ven/aza. In the propensity-matched cohort analyzing OS, older age, adverse risk, and RUNX1 mutations favored ven/aza over IC, whereas intermediate risk favored IC over ven/aza. In conclusion, patients receiving IC have improved OS compared with those receiving ven/aza. However, in a propensity-matched cohort of patients with equivalent baseline factors, there was a trend toward favorable OS for ven/aza. Specific variables, such as RUNX1 mutations, reported here for the first time, can be identified that favor ven/aza or IC, helping to guide treatment decisions for patients who may be eligible candidates for either therapy.

Classification of Acute Myeloid Leukemia Subtypes M1, M2 and M3 Using K-Nearest Neighbor

<p class='IJASEITAbtract'>Leukemia is a malignant disease caused by the massive and rapid development of white blood cells in the bone marrow. These excessive white blood cells begin to interfere with the body’s mechanism rather than fighting infection. Acute Myeloid Leukemia (AML) is one of the four main types of leukemia with eight subtypes, M0 to M7. AML M1, M2, and M3 have similarities, making them more difficult to distinguish from the other types. Furthermore, they are usually identified by calculating the ratio of myeloblast, promyelocyte, and monoblastic. This research aims to apply the k-Nearest Neighbor (k-NN) in classifying these cell types. k-NN is an algorithm used for classification based on a similarity measure. In cases of finding the best number of neighborhoods, trial and error were conducted. The features needed for classification are cell area, perimeter, roundness, nucleus ratio, mean and standard deviation. Four distance metrics such as Euclidean, Manhattan, Minkowski, and Chebyshev were used in this research. The results show that the Euclidean, Manhattan, Chebyshev, and Minkowski distance successfully identified 207 out of 300 objects at K=18, 197 out of 300 objects at K=13, 209 out of 300 correct objects at K=9, and 210 out of 300 objects at K=7. In conclusion, Minkowski was chosen as the best distance metric for KNN in classifying leukemia-forming blood cells. Furthermore, the accuracy, recall, and precision values of KNN with Minkowski distance obtained from 5-fold cross-validation were 80.552%, 44.145%, and 42.592%, respectively.

Simultaneous Occurrence Of Multiple Myeloma and Acute Myeloid Leukemia: A Dual Malignancy Of The Hematopoietic System

The occurrence of acute myeloid leukemia (AML) after the treatment for multiple myeloma (MM) is well described in several cases. The simultaneous presentation of MM and AML is very rare, with only 17 cases reported in articles found at PubMed.A 69 years old woman was sent at our institution for consultation due to pancitopenia at check-up. Past medical history was irrelevant. The CBC reported Hb 11.3 g/dL, Htc 33.8 %, ESR 32 mm/hr, Rouleaux formation +++, WBC 24.5 K/µL; differential showing nuetrophils 0.748 K/μL, lymphocytes 2.2 K/μL and myeloid blasts 20.1 K/µL, platelets 23.4 K/µL. The bone marrow aspiration was hypercellular with infiltration by myeloid blasts counting 95 %, and plasma cells with neoplastic morphology counting the remaining 5 %, no normal hematopoietic cells were observed. Hematological malignancy work up, also for MM, was performed, including flow cytometry (FC) and G band karyotype (GBK). FC revealed three populations: 1) representing 82.9 % that expressed CD11b +, CD13 ++, CD33 +, CD34 +, CD 38 +, HLA-DR + and MPO +; 2) representing 4.6 % and expressed CD38 +++, CD56 +, CD138 +++, kappa ++; 3) mixed population of normal B and T cells, representing 12.5 %. GBK showed 18 normal metaphases and 2 metaphases with t(11:14) (q13:q32) and del(13q). Stratification tests reported creatinin 2.3 mg/dL, , Ca 10.15 mg/dL, us-CRP 23 mg/dL, β-2-microglobulin 7.751 mg/L, albumin 2.35 g/dL, globulin 5.75 g/dL, serum protein electrophoresis with M-spike in the gamma region with 4.12 g/dL, positive Bence Jones protein, X-ray from the skull showed multiple lytic lesions. High risk MM and AML M4 were diagnosed. The patient did not accept any treatment and died few days later due a pulmonary hemorrhage.There is no pathophysiologic mechanism that explains the simultaneous occurrence. MM is a slowly disease, it may lead to chromosomal instability and immunosuppression with failure to eliminate clones of leukemic cells. No effective treatment is currently available, allogeneic stem cell transplantation is the best option. [Display omitted] [Display omitted] [Display omitted] Disclosures:No relevant conflicts of interest to declare.

Combinatorial Use of Bexarotene Plus ATRA for the Treatment of Acute Myeloid Leukemia

Aim: All-trans retinoic acid (ATRA) results in high response rates in acute promyelocytic leukemia (APL), but has not had a significant impact on non-APL forms of acute myeloid leukemia (AML), even though retinoic acid receptors have been implicated in hematopoietic maturation and self-renewal. RXRs bind as heterodimers with RARs, and are expressed at higher levels in AML cells than are RARs. We sought to determine whether natural RXR ligands exist in primary leukemia cells, whether RXR ligands might augment or inhibit leukemic cell growth, and whether RXRs might be rational targets for AML therapeutics.Background: Retinoid receptors are nuclear hormone receptors, induced during terminal myeloid maturation with highest expressed in M4 and M5 AML. Retinoids serve as intracellular messengers and are activating ligands for the retinoic acid receptors (RAR) and retinoid X receptors (RXR).Methods: We developed an in vivo reporter assay that measures intracellular, activating nuclear receptor ligands in mouse bone marrow cells. We generated transgenic mice with GFP regulated by a UAS promoter. The yeast Gal4 transcription factor recognizes UAS promoter sequences. When a fusion of the Gal4-DNA binding domain (DBD) and a retinoid receptor ligand-binding domain (LBD) is expressed in UAS-GFP bone marrow cells, and an activating ligand is present, the cells express GFP. Leukemia was generated in bone marrow cells from these mice and from Mx1-Cre x Rxrαfl/fl x Rxrβfl/fl mice using MSCV-MLL-AF9 retrovirus.Results: We found that intermediate levels of RXRA ligands are present in mouse MLL-AF9 leukemia cells in vivo. To determine if activation of RXRA is required for MLL-AF9 cell growth, we transduced MSCV-MLL-AF9 retrovirus into bone marrow cells from Mx1-Cre+ x Rxrαfl/fl x Rxrβfl/fl mice and transplanted these into recipient mice. Mx1-Cre is “leaky.” Without external activation of Mx1-Cre (i.e. without pIpC treatment) we found that all subsequent leukemias were associated with deleted (“floxed out”) Rxra and Rxrb alleles, suggesting that the deleted allele was positively selected for. In secondary transplants, we found that MLL-AF9 leukemias with deleted Rxrα and Rxrβ were associated with shorter survival when compared with MLL-AF9 leukemias with wild type Rxrα and Rxrβ, suggesting that naturally occurring RXR ligands suppressed leukemic growth. To determine if pharmacologic activation of retinoid receptors could inhibit MLL-AF9 cell growth, we treated MLL-AF9 cells ex vivo with ATRA and bexarotene. We found that ATRA modestly inhibited growth, and this effect reached a plateau at 62 nM (this may partially explain the limited clinical efficacy of ATRA in non-APL AML) (see Figure). In contrast, the EC50 of bexarotene was 125 nM and the EC50 decreased to 54 nM when bexarotene was combined with 62 nM ATRA. Higher doses of ATRA decreased the bexarotene EC50 further to 43 nM and resulted in near complete culture collapse. Finally, to determine whether combination bexarotene and ATRA inhibited cell growth by inducing differentiation or apoptosis, we examined cytomorphology, cell division by Celltrace Violet staining (similar to CFSE), and Annexin V. Unexpectedly, we observed no evidence of differentiation, but rather increased Annexin V associated with ATRA and bexarotene-induced growth arrest.Conclusion: RXRs act as tumor suppressors in MLL-AF9 murine leukemia cells in vivo, and the activation of RXR/RAR heterodimers with bexarotene and all-trans retinoic acid leads to synergistic apoptosis and growth arrest. Additional experiments are now being performed to test the efficacy of this dug combination in vivo and assess their potential use in elderly relapsed/refractory AML. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Clinical Characteristics and Treatment Outcome of Acute Myeloid Leukemia with APL-like Morphology and Uncommon RARA Fusion Variants

Background. Acute promyelocytic leukemia (APL) is a subtype of acute myeloid leukemia (AML) characterized by the translocation t(15;17) (q22;q21) generating the PML-RARA fusion gene. In rare cases of AML with APL like-morphology, RARA can be fused to an alternative partner other than PML (e.g. PLZF, NPM1 , NUMA1 , STAT5B, PRKAR1A and others) and this genetic heterogeneity accounts for the different sensitivity to targeted agents, such as all- trans retinoic acid (ATRA) and arsenic trioxide (ATO).Patients and methods. We collected data on 17 patients with AML characterized by variant RARA fusions, diagnosed between 2005-2017 from 13 institutions in US, Europe, Brazil, India and Australia. Of the 17 patients, 14 harboured PLZF-RARA, 1 PRKAR1A-RARA , 1 STAT5b-RARA rearrangements and 1 t(11;17)(q23;q21) with rearranged RARA.Results. Morphological data were compatible with classical M3 AML in all cases, and 38% presented with laboratory signs of coagulopathy. Flow cytometry was available in 12/17 cases and was compatible with classical APL, with 5/12 patients (42%) expressing CD56. Three cases were therapy-related AML due to previous exposure to radio-chemotherapy for solid tumors. Complete clinical-biological data are shown in Table 1. Most patients were adults (age range, 28-83 years), only two being younger than 18 years. Median WBC and platelet counts were 15.7 x 109/L (range, 2.18-248) and 69 x 109/L (range, 26-170), respectively, with 53% of patients presenting with WBC >10 x 109/L. Fourteen out of 17 patients received induction with ATRA + chemotherapy (CHT), either intensive or low-dose based on age and performance status. In addition, one patient received ATO combined with CHT and ATRA, 1 CHT alone and 1 ATRA alone. After induction, 11 patients achieved complete morphologic remission (CR) (of whom 2 after a second induction cycle); 2 are currently in partial remission (PR), 1 died early due to acute renal failure and 3 had resistant disease and received re-induction therapy that resulted in progressive disease in 2 and stable disease (SD) in one patient. Overall, the CR rate after induction therapy was 65%. Of the 13 patients who underwent consolidation therapy, 8 received ATRA+CHT, 2 CHT alone, 2 gemtuzumab ozogamicin (GO), and 1 azacytidine. Five of these patients underwent subsequent allogeneic hematopoietic stem cell transplant (HSCT) in first CR (3 from an HLA-matched sibling and 2 from matched unrelated donors). Four patients relapsed during follow-up (3 PLZF-RARA and 1 STAT5b-RARA ). Of these, 1 died of disease progression, 2 are alive in CR2 and CR4, respectively, and 1 is currently undergoing salvage therapy. With a median follow-up of 11 months (range: 1 - 86), 12/17 patients are alive (70.5%); of these, 9 are alive in CR, 2 are alive in PR and 1 alive with SD. Overall, five patients (29.5%) died, 3 from disease progression, 1 from transplant-related complications and 1 from acute renal failure during induction therapy.Conclusions. Despite the reported poor sensitivity of these RARA variants to standard APL therapy, this series suggests a relatively favorable outcome after treatment with ATRA combined with AML-like regimens including, in younger patients, allogeneic HSCT.The relative contribution of ATRA to achievement of CR and consolidation of remission remains to be further investigated.Table legend.*CR, complete remission; **mCR, molecular complete remission;+Res, resistant disease; °ED, early death;§PR, partial remissionATRA, All-trnas retinoic acid; IDA, idarubicin; DNR, daunorubicin; GO, gemtuzumab ozogamycin; ARA-C, cytaranine; L-DAC, low-dose cytarabine; HiDAC, high-dose cytarabine; MTZ, mitoxantrone; ETO, etoposide; ATO, arsenic trioxide;HSCT, hematopoietic stem cell transplantation;MUD, matched unrelated donor [Display omitted] DisclosuresCicconi:TEVA: Speakers Bureau. Montesinos:Celgene Corporation: Honoraria, Research Funding. Schwarer:Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; BMS: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Specialised Therapeutics: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau. Lo Coco:TEVA: Honoraria, Speakers Bureau; Novartis: Honoraria, Speakers Bureau; Lundbeck: Honoraria, Speakers Bureau.

NUP98-KDM5A Fusion Gene Is Not Exclusive to Acute Megakaryoblastic Leukemia and Is an Independent Predictor of Poor Prognosis

NUP98 rearrangements are recurrent genetic events in pediatric acute myeloid leukemia (AML). We identified NPU98-NSD1 as a recurrent poor risk abnormality (Hollink et al, Blood 2011). Another recurrent fusion is NUP98-KDM5A, which was found in pediatric acute megakaryocytic leukemia (AMKL) and was associated with poor clinical outcome (de Rooij et al, Nature Genetics 2017). To date, it is unclear whether NUP98-KDM5A exclusively occurs in AMKL or also in other FAB-subtypes of childhood AML. The aim of this study was to determine the incidence of NUP98-KDM5A in large pediatric AML cohorts, and to define the biological and clinical characteristics of this subgroup.The study comprised a COG and a European cohort of patients, and included 997 patients from COG trials 03p1 and 0531 (Gamis et al, Blood 2013) and 354 European pediatric AML patients, previously described by Balgobind et al (Haematologica 2011), NUP98-KDM5A rearrangements were detected either by paired-end RNA sequencing or RT-PCR. Patients with FAB M3 AML were excluded from this study. For comparison of clinical characteristics between NUP98-KDM5A positive and negative patients, the χ-squared and the Fisher's exact test were used. Event-free survival (EFS) and overall survival (OS) were estimated using the Kaplan-Meier method; the Cox proportional-hazard model was used for multivariate analysis. For this analysis, age, white blood cell count (WBC), cytogenetic risk group, allogeneic hematopoietic stem cell transplant (HSCT) as time dependent variable and study group (COG vs European cohort) were used as co-variables.NUP98-KDM5A rearrangements were detected in 31 out of 1351 patients (2.3%). There was no significant difference in prevalence between the COG and European cohorts. 41% of the NUP98/KDM5A+ patients were male, compared to 53% of the NUP98-KDM5A- patients (p=0.24). The median age of NUP98-KDM5A patients was significantly lower as compared to patients without this fusion gene (5.8 vs 9.4 yrs; p=0.03). There was no difference in WBC between NUP98-KDM5A positive and negative patients (13.1*109/l vs.25.2*109/l, respectively, p=0.13). NUP98-KDM5A occurred in all FAB types, with FAB M7 being most frequent (33% vs 8% in NUP98-KDM5A - patients; p<0.001).The 5-year EFS of NUP98-KDM5A + patients was 36.8 ± 18.8%, compared to 48.1 ± 2.8% in the NUP98-KDM5A - patients (p=0.12). The 5-year OS of NUP98-KDM5A + patients was 41.1 ± 19.7%, a value significantly lower compared to the NUP98-KDM5A - patients (63.9 ± 2.8%; p=0.01), this finding suggesting that NUP98-KDM5A+ patients are difficult to rescue when they relapse. In multivariate analysis, NUP98-KDM5A was an independent risk factor for OS, but not for EFS, with hazard ratios of 1.76 (p=0.045) and 1.58 (p=0.083), respectively. Non-M7 (n=19) patients had lower survival rates with EFS of 26.4 ± 21.7% and OS of 30.0 ± 22.3% compared to M7 (n=9) patients (EFS 55.6 ± 33.1% and OS 64.8 ± 33.1%). These differences were not statistically significant.This study shows that NUP98-KDM5A is an aberration that occurs in all FAB types and not just in AMKL. NUP98-KDM5A is an independent risk factor for OS but not for EFS and identifies a poor prognostic subgroup. Although there was a big difference in survival between M7 and non-M7 NUP98-KDM5A rearranged patients, this was not statistically significant, probably because of the low number of patients in the two cohorts. Overall, these data suggest to screen for the presence of NUP98-KDM5A rearrangements all children with non-APL AML irrespectively of their AML FAB subtype, and to stratify these patients in the high risk group. DisclosuresNo relevant conflicts of interest to declare.

Transcript-Level Dysregulation of BCL2 Family Genes in Acute Myeloblastic Leukemia

Simple SummaryThe BCL2 family comprises pro- and anti-apoptotic proteins whose cellular abundance, modifications, and interactions determine the cell fate. Its contribution to pathogenesis of acute myeloid leukemia (AML) was not widely studied and none of the studies published before presented a complex picture of BCL2 family gene expression. Therefore, we decided to analyze the AML transcriptome sequencing data to outline this picture and look for relations between the expression of particular BCL2 family members and the genes encoding interacting proteins, presence of mutations, and clinical features. Our findings should not only shed light on apoptosis- and oncogenesis-related processes, but may also be implemented in clinical practice. Prognostic significance, association with response to therapy and potential application in the selection of therapeutic targets are of particular importance.The expression of apoptosis-related BCL2 family genes, fine-tuned in normal cells, is dysregulated in many neoplasms. In acute myeloid leukemia (AML), this problem has not been studied comprehensively. To address this issue, RNA-seq data were used to analyze the expression of 26 BCL2 family members in 27 AML FAB M1 and M2 patients, divided into subgroups differently responding to chemotherapy. A correlation analysis, analysis of variance, and Kaplan-Meier analysis were applied to associate the expression of particular genes with other gene expression, clinical features, and the presence of mutations detected by exome sequencing. The expression of BCL2 family genes was dysregulated in AML, as compared to healthy controls. An upregulation of anti-apoptotic and downregulation of pro-apoptotic genes was observed, though only a decrease in BMF, BNIP1, and HRK was statistically significant. In a group of patients resistant to chemotherapy, overexpression of BCL2L1 was manifested. In agreement with the literature data, our results reveal that BCL2L1 is one of the key players in apoptosis regulation in different types of tumors. An exome sequencing data analysis indicates that BCL2 family genes are not mutated in AML, but their expression is correlated with the mutational status of other genes, including those recurrently mutated in AML and splicing-related. High levels of some BCL2 family members, in particular BIK and BCL2L13, were associated with poor outcome.

Microrna-211 - a Novel Oncogene in Acute Myeloid Leukemia

MicroRNAs are a class of small, non-coding RNA molecules that regulate gene expression by post-transcriptional silencing and regulate many critical cellular processes. In cancer, microRNAs are emerging as important biomarkers of disease and a subset of microRNAs have well-established roles as drivers of cancer development. MicroRNA-211 (miR-211) is an intronic microRNA encoded on human chromosome 15, and deregulated miR-211 expression has been previously demonstrated in melanoma and a range of other cancers. A role for miR-211 in the hematopoietic system, and/or in the development of acute myeloid leukemia (AML) however remains unexplored.Using microRNA expression profiling, across two independent pediatric AML cohorts, we are the first to identify that miR-211 expression is elevated in a subset of patients, from more than 2-fold and up to 50-fold above non-leukemic remission controls. High expression of miR-211 tends to be clustered in patients with cytogenetically normal and M5 AML subtypes. Using hematopoietic reconstitution experiments in mouse models, we show for the first time that enforced expression of miR-211 alone in hematopoietic stem cells is sufficient to drive a partially-penetrant myeloproliferative disease that phenotypically resembles acute myeloid leukemia. Affected mice have an expansion of immature myeloid cells, expressing CD11b and Gr-1 cell surface markers, with monoblastic cellular morphology. This is accompanied by severe anemia and thrombocytopenia, and dissemination of immature hematopoietic cells into the spleen, liver and extramedullary spaces of the bone marrow. We have also explored the potential co-operation between miR-211 and AML oncogenes. Strikingly, preliminary data suggests miR-211 can co-operate with activating RAS mutations to immortalize cells in vitro, and drive AML in vivo .To further explore the molecular mechanisms of miR-211 in AML, we used RNA sequencing and differential expression analysis to compare transcriptional changes induced by miR-211 overexpression in disease mice relative to empty vector controls. We have identified Lin28a as the most downregulated miR-211 gene target. Lin28a is a well-established target of miR-125b in AML and its knockdown leads to an analogous expansion of myeloid cells in vivo as observed in miR-211-dependent AML.Collectively, our results demonstrate a previously unidentified role for miR-211 as an oncogenic microRNA in the development of AML and suggests that miR-211 may potentially drive the development of AML by blocking myeloid differentiation through the repression of Lin28a . DisclosuresNo relevant conflicts of interest to declare.

Complex rearrangement in acute myeloid leukemia M2 with RUNX1/RUNX1T1 fusion involving chromosomes 8, 17 and 21

BackgroundThe translocation t(8;21)(q22;q22) is one of the most frequent chromosomal abnormalities associated with acute myeloid leukemia (AML) sub type M2. About 3–5 % of cases with additional chromosomal abnormalities, including structural and numerical ones, are reported to include a complex translocation t(8;21;N).Case presentationHere we report a chromosome rearrangement observed in a 19 years-old female diagnosed with AML-M2. When subjected to (molecular) cytogenetic analyses a complex three-way translocation involving chromosomes 8, 17 and 21 was detected, forming not a t(8;21;17) as one would expect. Real time-polymerase chain reaction analysis using 6 AML specific markers showed the presence of RUNX1/RUNX1T1 fusion gene transcripts identical to those found in classical translocation t(8;21) coupled with presence of FLT3-ITD mutation identified by fragment analysis.ConclusionsThe present case highlights importance of complex rearrangements rarely encountered in AML, suggesting that all involved regions harbor critical candidate genes regulating the pathogenesis of AML, leading to novel as well as well-known leukemia associated chromosomal aberrations.

Lymphoid markers predict prognosis of pediatric and adolescent acute myeloid leukemia

Acute Myeloid Leukemia (AML) is a complex and highly aggressive disease. To characterize the prognostic factors of pediatric patients with AML relapse, a retrospective cohort study was performed to collect data from children and adolescents, at a hematological oncology reference center, over 11 years. We selected 51 cases of the disease, diagnosed and treated uniformly, divided into two groups: with complete remission (n = 33; 65 %) and with relapse (n = 18; 35 %). The groups were homogeneous concerning demographic characteristics and hematological parameters at diagnosis. AML M3 was the most common subtype (n = 19; 37 %) and was associated with a good prognosis. The highest rate of relapse was with AML M0 (n = 3 of 5 patients; 60 %). The most predominant gene mutation, FLT3-ITD, did not influence the prognosis in our study. The complete remission group presented a higher mean frequency of positive cells for the granulocytic marker CD13a at diagnosis. In cases with AML relapse, CD36, CD4, CD7, and CD22 were the most expressed markers. Increase incidence of recurrence was associated with CD7 (HR 1.035; p = 0.003), CD4 (HR 1.032, p = 0.001) and CD22 (HR 1.042; p = 0.049). Our results highlight the importance of analyzing immunophenotypic markers to help predict the outcome of AML in children and adolescents.

Acute mastoiditis with facial paralysis as initial symptom of acute myeloid leukemia relapse: a rare case report

BackgroundExtramedullary involvement of leukemia usually occurs in the skin and central nervous system. Isolated leukemic involvement of the temporal bone is a very uncommon situation, and it is seen frequently concomitant to hematologic or molecular relapse.Case presentationAn 8-year-old male with acute myeloid leukemia (AML) M2 with t (8,21) (FAB classification) in remission for 2 years was referred to our clinic with severe ear pain, persistent ear discharge, and hearing loss with left facial paralysis for 4 days. Physical examination revealed purulent ear discharge with severe edema of the left ear canal. The temporal computed tomography revealed otomastoiditis. However, temporal magnetic resonance imaging demonstrated a 9 × 6 × 5 mm contrast-enhancing lesion in the internal acoustic canal and thickening in VIIth and VIIIth cranial nerves. Mastoidectomy was performed for exploration and biopsy. The histopathological results of the mastoid bone and soft tissue samples revealed a relapse of AML. One month after the first chemotherapy regime, ear discharge stopped and facial palsy recovered totally. However, the patient died 9 months after the diagnosis of relapse.ConclusionTemporal bone relapse should be suspected in AML patients with symptoms of otomastoiditis and facial palsy. Further investigations including radiological examinations, surgery, and biopsy for avoiding misdiagnosis and delay in treatment are needed.

The delta isoform of phosphatidylinositol-3-kinase predominates in chronic myelomonocytic leukemia and can be targeted effectively with umbralisib and ruxolitinib

Chronic myelomonocytic leukemia (CMML) is a myelodysplastic syndrome/myeloproliferative neoplasm overlap syndrome characterized by monocytic proliferation in the presence of dysplastic bone marrow changes, inflammatory symptoms, and propensity for transformation to acute myeloid leukemia (AML), with a poor prognosis and limited treatment options. Unlike the α and β isoforms, the phosphatidylinositol-3-kinase (PI3K)-δ signaling protein is predominantly expressed by hematopoietic cells and therefore has garnered interest as a potential target for the treatment of lymphomas and leukemias. We revealed a pattern of increased PIK3CD:PIK3CA ratio in monocytic M5 AML patients and cell lines, and this ratio correlated with responsiveness to pharmacological PI3K-δ inhibition in vitro. Because CMML is a disease defined by monocytic clonal proliferation, we tested the PI3K-δ inhibitor umbralisib as a single agent and in combination with the JAK1/2 inhibitor ruxolitinib, in CMML. Our ex vivo experiments with primary CMML patient samples revealed synergistic inhibition of viability and clonogenicity with this combination. Phospho-specific flow cytometry revealed that dual inhibition had the unique ability to decrease STAT5, ERK, AKT, and S6 phosphorylation simultaneously, which offers a mechanistic hypothesis for the enhanced efficacy of the combination treatment. These preclinical data indicate promising activity by co-inhibition of PI3K-δ and JAK1/2 and support the use of ruxolitinib + umbralisib combination therapy in CMML under active clinical investigation.

A Comparative Study of Digital Image Segmentation Algorithms for Acute Myeloid Leukemia M1 White Blood Cells Images

Various types of algorithms have been widely used for image segmentation in digital image processing. Every algorithm has features that make it unique to be applied to specific cases. One of the applications of image segmentation is to detect white blood cells. Certain objects such as blood cells must be able to be well segmented because their existence is very crucial to support the accuracy of disease detection related to haematology or the branch of medical science that studies the morphology of blood and blood-forming tissues. Three image segmentation algorithms were compared through this study: Seed Region Growing, Otsu Thresholding and Active Contour Without Edge. Comparative analysis of the three algorithms was done by counting the number of white blood cell objects that were successfully segmented with the actual number of cells that were counted manually. A total of 30 images of blood smears were taken from people suffering from acute myeloid leukemia M1. The average accuracy values from each algorithm were used to determine which image segmentation algorithm is the most suitable for application in the case of white blood cells segmentation. The results showed that Active Contour Without Edge is the most appropriate among the other algorithms

Image Segmentation of Acute Myeloid Leukemia Using Multi Otsu Thresholding

Acute Myeloid Leukemia (AML) can be identified by utilizing image processing. The stages used in the image processing process include preprocessing, segmentation, and feature extraction. The purpose of this segmentation is to separate objects in the image. One of the image segmentation methods is Multi Otsu Thresholding. This method has a way of working by looking for the threshold value dynamically. Previously there has been researched using the Static Otsu Thresholding method, but the results are still unsatisfactory because the threshold value obtained in the Static Thresholding method is that the threshold value obtained is static, meaning that the threshold value cannot adjust to the brightness level of each image that will be identified. With the background of the segmentation result dissatisfaction factor from the static threshold value, this study aims to carry out the segmentation process to obtain a dynamic threshold value, namely the Multi Otsu Thresholding Method. The Multi Otsu Thresholding method will be applied to segment AML images of M0 and M1 types, with the hope that the segmentation results obtained can be used for the feature extraction process better and are useful for the identification and classification process. Image processing methods used in this research are YCbCr color space, median filter, multi otsu thresholding, and morphological operations. The cell identification process utilizes the cell type classification process using the Naïve Bayes Classifier. The extracted characteristics were WBC (White Blood Cell) diameter, nucleus ratio, and nucleus roundness. Image data to test the process are 29 images of AML M0 and 30 images of AML M1. It was found that the segmentation results using the Multi Otsu Thresholding method can be used for the feature extraction process in AML M0 and AML M1 images and used for the identification process using the Naïve Bayes Classifier resulting in an accuracy of 83.81%, in this case, it has been compared with Static Otsu Thresholding which results with an accuracy of 75.35 %.

Periodontist an Asset to Systemic Diagnosis — Diagnosing a Case of Acute Myeloid Leukaemia

Acute Myeloid Leukemia (AML) is a neoplastic disease of bone marrow. AML M3 variant frequently develops swiftly and if not intervened immediately may prove fatal. This report presents a 32-year-old female who reported to dental OPD with complaint of bleeding gums and no previous medical history. On examination suspicious lesions were seen and patients was advised for full blood workup and Peripheral Blood Smear (PBS) investigations which revealed features suggestive of AML M3. The significant findings of this case are just presentation of oral manifestations, quick progression from initial symptoms to end stage and mild increase in WBC count which is uncommon for AML M3 variant. For many types of cancer, finding it early might make it uncomplicated to treat. This report outlines how a periodontist proved to be an asset in diagnosing a case of AML.

Diagnóstico ocasional de leucemia mielóide aguda em vigência de infecção por SARS-CoV-2

OBJECTIVES: To describe a patient with SARS-CoV-2, who was diagnosed with acute myeloid leukemia M3. CASE REPORT: Patient, 12 years old, female, started with bruises in the lower limbs and pain in the left leg; however, in the face of the current pandemic, the patient was delayed in seeking medical help due to fear of contamination by SARS-CoV-2. After 1 month she presented fever, runny nose, and cacosmia. She sought medical assistance for apprehension of being infected with the new coronavirus due to the fact that the mother is a health professional. The child was admitted to the emergency room, with leukopenia, thrombocy-topenia, and positive COVID-19. She remained hospitalized for 8 days in the hospital of entry. Initially, it was suspected of idiopathic thrombocytopenic purpura; however, with the evolution of the case it suggested the diagnostic possibility of oncological disease. She was transferred to a specialty hospital, where she was diagnosed with acute myeloid leukemia subtype M3. After diagnosis, tretinoin associated with platelet transfusion and cryoprecipitate and subsequent chemotherapy with idarubicin was started. CONCLUSIONS: It can be analyzed that, in the face of the SARS-CoV-2 pandemic, many diseases can arise together with the viral infectious condition. On the other hand, we cannot fail to consider that in view of the current epidemiological and behavioral changes, the diagnosis, treatment, and prognosis of previously existing diseases that, in due course, coexist with SARS-CoV-2 infection, can be postponed, due to the fear and due to the difficulty of accessing the health system.

Role of homeobox-A9 gene expression in patients with de novo acute myeloid leukemia

ObjectiveTo study the clinical significance and the prognostic value of homeobox-A9 (HOXA9) gene in patients with newly diagnosed acute myeloid leukemia (AML).BackgroundHOX genes are transcription factors. In humans, the 39 HOX genes are organized into four genomic regions (the HOXA, B, C and D clusters), located on four chromosomes (chromosomes 7, 17, 12, and 2, respectively). They play important roles during embryogenesis. The characteristic expression of HOX genes can also be detected in different adult tissues.Patients and methodsThis prospective case–control study was conducted on 60 patients with newly diagnosed AML and 40 healthy patients serving as a control group. They underwent a full history, clinical examination, complete blood count, bone marrow examination, and quantification of HOXA9 in blood by quantitative real-time PCR assay.ResultsFrom the results of this study, we found that HOXA9 was higher in patients with AML than healthy controls (P < 0.001). The highest incidence was found in AML M7 and M5, in patients with AML with older age, with hepatomegaly and splenomegaly, in patients with AML with unfavorable cytogenetic criteria, and in patients with AML who did not achieve complete remission.ConclusionHOXA9 is higher in patients with AML than controls. High HOXA9 expression has a negative prognostic effect on patients with AML and poorer response to chemotherapy. Moreover, it represents an independent prognostic factor mediating chemotherapy.

Acute myeloid leukemia M1 FAB secondary to myelodysplastic syndrome with excess blasts type 2

Acute myeloid leukemia occurs naturally in the evolution of myelodysplastic syndrome (MDS), typically in MDS with excess blasts type 2. The risk of progression is around 33%. We present the case of an elderly patient who was diagnosed with MDS single dysplasia in 2016. He was treated with erythropoietin. In 2021, the disease evolved into MDS with excess blast type 2 and then in acute myeloid leu­ke­mia in a short period of time.