Clinical Features of DIC According to the French-american-british (FAB) Classification in Patients With Acute Leukemia and Thrombomodulin Alfa Treatment – Cross-sectional Analysis of a Post-marketing Surveillance Study Cohort –

Background: The aims of this study were to analyze the clinical features of a large number of cases with disseminated intravascular coagulation (DIC) associated with acute leukemia, and to clarify the safety and efficacy of thrombomodulin alfa (TM-α) using the French-American-British (FAB) classification of hematological malignancies.Methods: We retrospectively examined 644 patients with acute leukemia in post-marketing surveillance for TM-α.Results: M3, M2, M4, M1, and M5 subtypes of acute myeloid leukemia (AML), and L2 and L1 subtypes of acute lymphoblastic leukemia (ALL) have been found more frequently among patients with DIC. Bleeding symptoms at baseline were more frequent in M3 and M7 subtypes. Fibrinogen concentrations were lower and plasmin-plasmin inhibitor complex values were higher in M3 and Philadelphia-positive (Ph+) ALL. Overall DIC resolution rate was 60.2% with higher rates in L1 and Ph+ ALL, lower in M1, and generally higher in ALL than in AML. The overall survival rate was 79.8%, generally high, with higher survival rates in L3, Ph+ ALL, and M3. In M3 and M7, with high frequencies of pre-existing bleeding, TM-α improved bleeding symptoms. Post-administration DIC scores in each subtype were significantly improved compared with pre-administration scores, except in M6, M7, and MDS-overt AML.Conclusions: This study showed the clinical features of DIC associated with acute leukemia among FAB classifications and also elucidated the safety and efficacy profiles of TM-α by detailed classification based on the FAB classification in clinical practice.Trial registration: The clinical characteristics and treatment outcomes of patients with DIC treated with TM-α between May 2008 and April 2010 were retrospectively analyzed by subgroup analysis of post-marketing surveillance data.

Study of the Age Dynamics of Morphological Subtypes of Acute Myeloid Leukemia in Adults

Aim: to determine the age dynamics of morphological subtypes of acute myeloid leukemia in adult patients. Materials and methods. The study group consisted of 132 patients (including 34 aged 15 to 45 years, 46 aged 45-60 years, 52 aged over 60 years) with newly diagnosed AML. Patients with acute promyelocytic leukemia were not included in the study. The diagnosis was made in accordance with WHO recommendations and FAB classification criteria. In all cases, morphological verification was performed, including cytological, cytochemical studies and immunophenotyping. Detection of chromosomal abnormalities was performed using standard cytogenetic and real-time polymerase chain reaction methods. Point mutations were screened in 7 genes: c-KIT, DNMT3A, FLT3, NRAS, NPM1, TP53 and WT1 by direct sequencing method. Results. The distribution of morphological variants of AML according to the FAB classification was as follows: M2 — 47.7%, M4 — 26.5%, M1 — 7.6%, M0 — 4.5%, M6 — 4.5%, M5 — 3.0%, M4eo — 2.3%, blastic plasmacytoid dendritic cell neoplasm — 1.5%, acute myelofibrosis — 0.8%, acute hybrid leukemia — 0.8%. Complex genetic abnormalities were detected in 18.2%, specific chromosome abnormalities associated with a favorable prognosis — 7.6%, specific chromosome abnormalities associated with an unfavorable prognosis — 5.3%, normal cytogenetics — 28.8%. Mutations in the FLT3 gene were detected in 15.0%, NPM1 — 14.3%, DNMT3A — 12.5%, NRAS — 12.2%, TP53 — 9.6%, c-KIT — 5.7%, WT1 — 4.2%. It was revealed that the frequency of acute myeloid leukemias with monocytic differentiation (M4, M4eo and M5 according to FAB) had a statistically significant tendency to decrease with increasing age: in the subgroup of patients younger than 45 years, it was 44.1%, in the age of 45-60 years — 32.6%, in the age of 60 years and older — 23.1%. These differences correlated with the age-related dynamics of the frequency of genetic abnormalities in AML, including a decrease in the frequency of NPM1 gene mutations and specific chromosomal aberrations, as well as an increase in the frequency of TP53 abnormalities in older age groups.

Interactions of Consanguinity and Number of Siblings with Childhood Acute Lymphoblastic Leukemia

Acute lymphoblastic leukemia (ALL) is a common malignancy in children. Consanguinity has a high prevalence in developing countries and increases the probability of homozygosity for many genes which may affect ALL and its prognosis. We conducted a study to explore the impact of consanguinity and number of siblings on ALL as there are currently no studies to describe this effect. Data were collected from patients’ records from the Children’s University Hospital of Damascus University, which is the major cancer centre for children in Syria. This study included 193 children with ALL over one year. Number of siblings was not with the French–American–British (FAB) classification, gender, ALL subtype, or risk of ALL children. When comparing consanguinity degrees and complete blood counts at diagnosis, significant contradicting data were found in the third-degree and fourth-degree consanguinity when compared to one another and to not having consanguineous parents as third degree consanguinity was associated with normal platelets but lower WBC counts, and fourth-degree consanguinity was associated with normal haemoglobin levels and WBC counts, but lower platelet counts. Having consanguineous parents was also associated with acquiring ALL at an older age, L2 FAB classification, having a positive family history for malignancies, and not having hepatosplenomegaly ( P < 0.05 ). Although L2 is known to be a poor prognosis indicatory, no association was found with consanguinity and risk. Finally, no association was found with ALL subtype or risk ( P > 0.05 ). Although consanguinity and number of siblings have affected some variables and prognostic features of childhood ALL, the aetiology is not clear and we need further studies to clarify such an association as this will help in optimising therapy and accurately determine the risk.

Association between the CEBPA and c-MYC genes expression levels and acute myeloid leukemia pathogenesis and development

CEBPA and c-MYC genes belong to TF and play an essential role in hematologic malignancies development. Furthermore, these genes also co-regulate with RUNX1 and lead to bone marrow differentiation and may contribute to the leukemic transformation. Understanding the function and full characteristics of selected genes in the group of patients with AML can be helpful in assessing prognosis, and their usefulness as prognostic factors can be revealed. The aim of the study was to evaluate CEBPA and c-MYC mRNA expression level and to seek their association with demographical and clinical features of AML patients such as: age, gender, FAB classification, mortality or leukemia cell karyotype. Obtained results were also correlated with the expression level of the RUNX gene family. To assess of relative gene expression level the qPCR method was used. The expression levels of CEBPA and c-MYC gene varied among patients. Neither CEBPA nor c-MYC expression levels differed significantly between women and men (p=0.8325 and p=0.1698, respectively). No statistically significant correlation between age at the time of diagnosis and expression of CEBPA (p=0.4314) or c-MYC (p=0.9524) was stated. There were no significant associations between relative CEBPA (p=0.4247) or c-MYC (p=0.4655) expression level and FAB subtype and mortality among the enrolled patients (p=0.5858 and p=0.8437, respectively). However, it was observed that c-MYC and RUNX1 expression levels were significantly positively correlated (rS=0.328, p=0.0411). Overall, AML pathogenesis involves a complex interaction among CEBPA, c-MYC and RUNX family genes.

Studies on the morphology of leukaemic blast cells in relation to haematological parameters

A combination of haematological parameters with morphological evaluation of peripheral blood and bone marrow blast cells is crucial for leukaemia diagnosis. FAB (French– American–British) classification is a simple and powerful diagnostic tool for leukaemia in developing countries like India. Differentiation block in the early stages of haematopoiesis and morphological characteristics of leukemic blast cells are directly related to haematological parameters. The present study is an approach to increase understanding of the simple morphological FAB classification of leukaemia in relation to haematological parameters. The present study revealed that Chronic Myeloid Leukaemia (CML) was the most common type of leukaemia , followed by Acute Myeloid Leukaemia, Acute Lymphoid Leukaemia (ALL), and Chronic Lymphoid Leukaemia (CLL) in Nagpur. Most of the cases of Acute Leukaemia had severe anaemia and thrombocytopenia. Highest variation was found in Total WBCs count of different types of leukaemia , particularly in different subtypes of AML. The present study also suggested that FAB classification is not outdated, but it does require continuous revalidation and other procedures for refinement.

The results of treatment in children with AML and initial hyperleucocytosis according to the AML-MM-2006 Protocol

The prognostic role of HL in AML in children is a matter of a discussion. 185 patients were treated for AML in our center, 36 of 185 had HL (19.5%). The study was approved by the Independent Ethics Committee of the Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology, and Immunology. Median Le was 97 × 109 /l (50–428 ± 109 /l). Standard risk group were 4 pts, intermediate – 8, hight – 24. The most common variants were M4/M5 in FAB classification – 30 pts and genetic rearrangement – MLL in 15 of 36 pts. Thirty-five patients with HL received cytoreduction course and ADE. After that, all patients received ADE and 21 pts second part of induction – course HAM. Remission was achieved in 27 (75%) out of 36 pts. HSCT was performed in 23 pts. Thirteen out of 36 patients with HL died: 4 (30%) – due to leukostasis complications. OS for HL group was 0.56 ± 0.09, for non-HL group was 0.75 ± 0.04, p = 0.005; EFS (HL) 0.42 ± 0.09, EFS (non-HL) 0.49 ± 0.04, p = 0.026. Also, differences in I CR achievement, median of remission length and death before remission between two groups were statistically significant (p = 0.036; p = 0.028; p = 0.021 respectively). OS and EFS in patients with M4/M5 with HL > 50 ± 109 /l were better than in patients all FAB with HL > 100 ± 109 /l, OS 0.71 ± 0.1 vs OS 0.43 ± 0.1 (p = 0.012); EFS 0.54 ± 0.1 vs EFS 0.29 ± 0.1 (p = 0.038) respectively. HL significantly worsens OS and EFS in children with AML.

Acute Non Lymphoblastic Leukemia in the Department of Child Health School of Medicine, University of North Sumatera/ Dr. Pirngadi Hospital Medan (1983-1988): A Preliminary Study

A retrospective study on Acute Non Lymphoblastic Leukemia (ANLL) was conducted at the Sub Division of Pediatric Hematology, School of Medicine, University of North Sumatera/Dr. Pirngadi Hospital Medan, in a period of 5 years (1983-1988). There were 18 cases consisted of 14 (77. 78%) males and 4 (22 .22%) females with the age group of 0-2 years: 6 (30%), 2-8 years: 9 (50%), 8-15 years: 3 (30%). By the FAB classification, they were of FAB M-1: 1 (5.55%), FAB M-2: 1 (5.55%), FAB M-3: 1 (5.55%), FAB M-4: 2 (11.12%) and FAB M-6: 13 (72.23%). Only 7 (38.88%) were treated with cytostatics while the others received only supportive therapy. The result of cytostatic treatment was unsatisfactory: 4 (57.14%) died within the first 2 months of treatment, 3 (42.86%) discontinued their cytostatics treatment.

Characterization of Mutations in the Mitochondrial Encoded Electron Transport Chain Complexes in Acute Myeloid Leukemia

Background: Acute Myeloid Leukemia (AML) is a devastating and heterogeneous, hematological malignancy characterized by uncontrolled proliferation of undifferentiated myeloid progenitor cells-blasts. AML cells are highly oxidized compared to healthy hematopoietic stem cells; thus, mechanisms contributing to this oxidation state represent an opportunity for targeted therapy. Oxidative phosphorylation (OXPHOS) is a key intracellular process where electrons are passed through the Electron Transport Chain (ETC) to produce the cellular fuel-ATP; however, during this process electrons leak from certain complexes of the ETC to form superoxide anions-free radicals that bind to iron-sulfur cores of important enzymes and inactivate them, inhibiting pivotal cellular pathways. Mutations or defects in the mitochondrial encoded ETC complexes (I, III, IV and V) could have significant effects on the metabolic balance of the mitochondria. While mitochondrial DNA (mtDNA) mutations have been previously reported in hematologic malignancies, including AML, mutations in the ETC genes encoded by the mitochondria have not been fully characterized. Methods: We analyzed molecular and clinical patient data retrieved from the Cancer Genome Atlas (TCGA) dataset for 200 patients with AML. We assessed the association between the presence of mutations in the mitochondrial encoded ETC with the clinical and molecular characteristics of patients with AML, using the Mann-Whitney U or Fisher Exact test. Additionally, we generated Kaplan-Meier survival curves to assess the association between presence of mutations in mitochondrial encoded ETC genes and clinical outcome. For multivariable survival analysis, we used the Cox Proportional Hazards Model to adjust for other risk factors. All statistical analysis was done using STATE 12.0 SE. Results: We found that mtDNA ETC mutations were present in 8% of patients with AML (16 of 200) with most of the mutations present in Complex IV (9 of 16 patients: 56.3%). MtDNA ETC mutations were significantly more frequent in older patients (%: 81.2 vs 18.8; p-value: 0.003), in patients with M0 FAB classification (%: 26.7 vs 8.22; p-value: 0.04) and in patients with poor molecular risk (%: 50 vs 23.4; p-value: 0.035). The median age of patients with Complex IV mutations was higher than that in patients without the mutations (median: 72 vs 57; p-value: 0.009), and these mutations were more frequent in patients with M0 FAB classification (%: 33.3 vs 8.47; p-value: 0.04). TP53 mutations were significantly more frequent in patients with mtDNA ETC gene mutations (%: 31.3 vs 5.95; p-value: 0.004). Similarly, after complex stratification, mutations in TP53 were more frequent in patients with Complex IV mutations (%: 33.3 vs 6.81; p-value: 0.026) and in patients with mutations in either or both Complex I and IV (%: 33.3 vs 6.38; p-value: 0.009). Patients with mtDNA ETC gene mutations had worse overall survival than wild type patients (median: 9.3 vs 20.1 months; p-value: 0.007). Comparably, after excluding M3 APL patients (due to their different treatment and improved overall survival), patients with mtDNA ETC mutations still had worse overall survival than patients carrying the mtDNA ETC wild type (median: 9.3 vs 15.8; p-value: 0.0139). Patients with Complex I mutations had worse overall survival (median: 5.85 vs 18.5 months; p-value: 0.009) than those without Complex I mutations. Likewise, patients with Complex IV mutations also had worse overall survival than those without Complex IV mutations (median: 7.0 vs 18.5 months; p-value: 0.047). In patients who did not receive transplant as part of their treatment regimen, presence of mtDNA ETC gene mutations were significantly associated with worse overall survival (median: 4.35 vs 9.9 months; p-value: 0.0123). And among TP53 mutated patients, mutations in either or both Complex I and IV were also significantly associated with shorter overall survival (median: 0.85 vs 9.4 months; p-value: 0.008). Conclusion: Mutations in mitochondrial encoded ETC genes were more frequent in older patients and were significantly associated with mutations in TP53 and shorter overall survival. Elucidation of the mechanisms by which ETC mutations contribute to AML pathogenesis and progression will facilitate the development of novel therapeutic targets. Disclosures No relevant conflicts of interest to declare.