et al. NPM1 but not FLT3-ITD mutations predict early blast cell clearance and CR rate in patients with normal karyotype. AML(NK-AML) or high-risk myelodysplastic syndrome (MDS). Blood. 2009;113(21):5250-3. 20. Gaidzik VI, Schlenk RF, Moschny S, Becker A, Bullinger L, Corbacioglu A, Krauter J, Schlegelberger B, Ganser A, Döhner H, Döhner K; German- Austrian AML Study Group. Prognostic impact of WT1 mutations in cytogenetically normal acute myeloidleukemia: a study of the German- Austrian AML Study Group. Blood. 2009;113(19):4505-11.
Figure 1. BCR-ABL downregulates PTEN expression. A) Left panel: evaluation of PTEN levels of expression in BCR-ABL-pcDNA3.1 transfected NIH3T3. Right panel: quantification of PTEN levels. B) PTEN mRNA levels in BCR-ABL transfected NIH3T3 cells. C) Left panel: PTEN levels by western immunoblot in primary CD34 positive cells obtained from the bone marrow of normal individuals and Chronic MyeloidLeukemia patients. Right panel: quantification of PTEN levels. D) Stably expressing pBabe-BCR-ABL infected NIH3T3 cells were treated with MG132 and UO126 for 8 hours. Control: parental NIH3T3 expressing pBABE-empty vector.
16. Byrd JC, Mrózek K, Dodge RK, Carrol AJ, Edwards CG, Arthur DC et al. Pretreatment cytogenetic abnormalities are predictive of induction of relapse, and overall survival en adult patients with de novo acute myeloidleukemia: results from Cancer and Leukemia Group B (CALGB 8461). Blood. 2002;100(13):4325-36. 17. Pelloso LAF, Chauffaille MLLF, Ghaname FS, Yamamoto M, Bahia
phocytes and 46% blast cells). A bone marrow aspirate showed hypercelullarity with 78% of myeloblasts and monocyte lineage cells. Cytochemical tests showed 44% Sudan-Black positive and 10% periodic acid Schiff (PAS) positive blast cells and 32% of non-erythroid blast cells were positive for non-specific alpha-naphylacetate este- rase. The patient reported no previous exposure to cyto- toxic or mutagenic agents. The diagnosis of de novo acute myeloidleukemia FAB classification type M4 was made. This study was approved by the ethics committee of the IBILCE-UNESP, São José do Rio Preto, SP, Brazil.
The importance of diagnostic cytogenetics on outcome in AML: analysis of 1,612 patients entered into the MRC AML 10 trial. Blood. 1998; 92(7):2322-33. 5. Slovak ML, Kopecky KJ, Cassileth PA, Harrington DH, Theil KS, Mohamed A, et al. Karyotypic analysis predicts outcome of preremission and postremission therapy in adult acute myeloidleukemia: a Southwest Oncology Group/ Eastern Cooperative Oncology Group Study. Blood. 2000; 96(13):4075-83. 6. Jourdan E, Boiron J-M, Dastugue N, Vey N, Marit G, Rigal-Huguet F, et al.
Chronic myeloidleukemia (CML) is a hematopoietic stem cell malignancy characterized by the t(9;22) chromosomal translocation that generates the BCR/ABL oncogene . BCR-ABL tyro- sine kinase inhibitors (TKI) are able to induce remission in CML patients but not to eliminate leukemia stem cells (LSC), which can regenerate leukemia on drug discontinuation [2–4]. Understanding LSC regulation is critical to understand CML pathogenesis and to develop cura- tive strategies. Proliferation, survival and drug-resistance of leukemic cells are largely dependent on their interplay with the bone marrow (BM) microenvironment, in which mesenchymal stem cells (MSC) are important components. Indeed, the functional MSC behavior is essential to favor or impede LSC expansion and, for this reason, MSC represent a possible target for treat- ment of leukemias . Since BM is a store of undifferentiated MSC, tumor cells precursors may affect the differentiation of MSC in the tumor niche suggesting a deep cross-talk between LSC and MSC . Interestingly, despite MSC from CML patients do not express BCR–ABL , recent studies have reported an altered regulation of MSC in CML, showing that changes in BM microenvironmental function suppress normal hematopoietic stem cells (HSC) and provide a selective advantage to LSC. Into the tumor milieu, MSC also play an important role for their immunosuppressive ability that can interfere with the immune recognition of tumor cells. Indeed, they produce and release immunoregulatory factors, including transforming growth factor β (TGF-β), prostaglandin E2 (PGE2), tumor necrosis factor α (TNFα), indolamine 2,3-dioxygenase (IDO), hemeoxygenase (HO), nitric oxidase synthase 2 (NOS2), arginase 1–2 (ARG1-2) and IL10 [5, 9–11]. MSC express programmed death ligand 1 (PD-L1) that after its engagement with PD-1 expressed on T lymphocytes leads to the inhibition of T cell activation and proliferation with an inefficient immune response .
Chronic myeloidleukemia (CML) is a clonal dis- ease characterized by the presence of the hybrid gene BCR/ ABL, usually located on the Philadelphia chromosome which results from reciprocal translocation between chro- mosomes 9 and 22. Two principal types of mRNA (b2a2 and b3a2) can be produced from this gene. These tran- scripts, which can also be co-expressed by alternative splic- ing, differ by the presence of BCR exon 14 (exon b3 on the hybrid gene). The 210-kDa protein translated from these mRNAs has increased tyrosine kinase activity, which plays a crucial role in leukemogenesis (Shtivelman et al., 1985; Lugo et al., 1990).
Acute myeloidleukemia (AML) is the most common acute leukemia in adults. The disease is characterized by various cytogenetic and molecular abnormalities with distinct prognoses and gene expression profiles. Emerging evidence has suggested that circulating microRNAs (miRNAs) could serve as noninvasive biomarkers for cancer detection; however, little is known about circulating miRNA profiles in AML patients. In this study, a genome-wide serum miRNA expression analysis was performed using Solexa sequencing for initial screen, followed by validation with real-time PCR assays. The analysis was conducted on training and verification sets of serum samples from 140 newly diagnosed AML patients and 135 normal adult donors. After a two-phase selection and validation process, 6 miRNAs, miR-10a-5p, miR-93-5p, miR-129-5p, miR-155-5p, miR- 181b-5p and miR-320d, were found to have significantly different expression levels in AML compared with control serum samples. Furthermore, unsupervised clustering analysis revealed the remarkable ability of the 6-miRNA profile to differentiate between AML patients and normal controls. The areas under the ROC curve for the selected miRNAs ranged from 0.8129 to 0.9531. More importantly, miR-181b-5p levels in serum were significantly associated with overall survival. These data demonstrated that the expression patterns of circulating miRNAs were systematically altered in AML and miR- 181b-5p may serve as a predictor for overall survival in AML patients.
Acute myeloidleukemia (AML) is a disease predominantly of older adults. Treatment of AML in the elderly is complicated not only by comorbidities but also by the high prevalence of poor prognosis markers. Thirty-one consecutive unselected patients with AML older than 60 years (representing 33% of all AML cases diagnosed at our institution during the same period) were followed over a period of 5 years (1997-2002). A high incidence of AML with multilineage dysplasia (45%) and no favorable cytogenetic abnormalities but 62% intermediate and 38% unfavorable karyotypes were found. Sixteen patients (52%) were selected for induction of intensive cytotoxic treatment and complete remission was achieved only by some of these intensively treated patients (7 of 16). Of these, 3 remained alive without disease (median: 11 months), 1 patient died shortly after complete remission, and 3 patients relapsed and died from refractory disease. Only 1 patient that was refractory to intensive cytotoxic treatment remained alive with disease under supportive care. Fifteen patients (48%) were managed with palliative/supportive care: 7 re- ceived palliative treatment and supportive care, 8 received supportive care only, and 4 patients remained alive with disease under supportive care (median: 9 months). Mortality rate was 74% and overall survival at two years was 12%. To the best of our knowledge, there is no previous report regarding elderly patients with AML in Brazilian subsets. The present data are similar to previously reported studies showing that elderly AML patients are not only older but also biologi- cally distinct from younger AML patients, particularly in terms of the high incidence of poor prognostic karyotypes and resistance to therapy. Correspondence
Introduction. Skin findings in leukemias may be divided into specific lesions (leukemia cutis) and non-specific lesions (leu- kemids) which may be found in up to 80% of all patients with leukemias. The leukemids vary clinically and they are usually a manifestation of bone marrow or immunologic impairment, but also Sweet syndrome, pyoderma gangrenosum, erythro- derma, maculopapular exanthema, prurigo-like papules, gen- eralized pigmentation, follicular mucinosis, generalized pru- ritus may be found during the course of leukemia. Case re- port. We report a 70-year-old male with a 3-month history of erythema, papules and pustules on the face, ears and neck and over a month history of refractory anemia, anorexia, weight loss, malaise, and fever. Physical examination revealed sym- metric erythematous, violaceous papules, papulo-nodules and plaques with slate scale and sparse, small pustules on the face, earlobes and neck. Histopathologic findings of involved skin showed diffuse mixed inflammatory cell infiltrate with peri- follicular accentuation and focal granulomatous inflammation in the papillary and upper reticular dermis. Extensive check- up revealed the presence of acute myeloidleukemia French- American-British (FAB) classification subtype M2, with signs of three-lineage dysplasia. The patient was treated by L6 protocol which led to complete remission, both in bone mar- row and skin, but after seven months he had relapse of leu- kemia with the fatal outcome. Conclusion. This case indi- cates the importance of skin eruptions in the context of he- matological malignancies.
Chronic myeloidleukemia (CML) was the first neoplastic disease to be linked to a genetic abnormality and is the best studied molecular model of leukemia. The increased and cumulative knowledge of this disease has recently received special attention due mainly to the new therapeutic option, a signal transducer inhibitor, the imatinib mesylate.
Worldwide, an estimated 57 000 cases of leukemia occur every year  and acute myeloidleukemia (AML) is the most common acute leukemia (AL). The highest incidence rate is found in males of all age groups, the fact remains to be explained [2–4]. The etiology of most types of leukemia is still unknown. Leukemia is likely to be associated with certain environmental agents, such as ionizing radiation, benzene, and cancer chemotherapy. The increase risk factors for leukemia may be both quantity and quality changes in folic acid metabolism [5–7].
Figure 4.1 Concept of this lab-on-chip as a point-of-care application: i) White cells extracted from a small blood sample are collected to analyze gene expression; ii) This approach aims to diagnose chronic myeloidleukemia using its genetic marker, BCR-ABL1 fusion transcript; iii) Total RNA extracted from white blood cells is then mixed with Au-nanoprobes and heated to promote hybridization. Note that Au- nanoprobes are functionalized with BCR-ABL1 complementary sequences; iv) The resulting solution and a salt solution are infused on the two microfluidic chip inlets; v) Thorough mixing and optical detection of these components is performed inside the microfluidic chip. If the patient expresses BCR- ABL1 transcripts complementary to the oligonucleotide sequence of Au-nanoprobes, their hybridization will cause the final solution to remain red (positive match) in the presence of salt. Otherwise, the non- hybridized Au-nanoprobes will aggregate and cause the final solution to turn blue (negative match) in the presence of salt. With the appropriate setup, and using LabView software, output results described in this step are displayed on the computer within 3 minutes.
overshadowed in acute leukaemia due to increased incidence of bleeding and infections. Acute MyeloidLeukemia is a morphologically and genetically heterogenous disease. One subtype with specific morphology and specific cytogenetic and molecular genetic aberration is acute promyelocytic leukemia (APL). On the basis of morphology alone, APL can be separated into two distinct subtypes according to FAB classification: AML M3 and AML M3 variant (AML M3 v).The latter is also called microgranular (hypogranular) APL in new WHO classification. The cytomorphology of APL blasts is obviously different in the two subtypes: in AML M3, the abnormal promyelocytes show heavy granulation and bundles of Auer rods; in AML M3v, blasts have agranular or hypogranular cytoplasm or contain fine dust like cytoplasmic granules that may sometimes be unclear on light microscopy. Furthermore, M3v blasts show a typically bilobed nuclear configuration. This latter morphology, together with missing granulation, often resulted in a misleading diagnosis of acute monocytic or myelomonocytic leukemia 6 .Flowcytometry of
The aim of this study is to evaluate the effects of Nucleophosmin1 mutation on the clinical presentations, prognosis, diagnosis and the treatment of acute myeloidleukemia. Thirteen articles were extracted from PubMed, Google scholar and Scopus in which the Nucleophosmin1 mutation correlated with gingival hyperplasia, high white blood cell count, lymphadenopathy, high platelet count and other signs and symptoms of myelomonocytic and monocytic acute myeloid leukemias.
QCM and EIS techniques have also been used to characterize lectin –cell interactions, including cell-surface glycosylation [32,33] or discrimination among malignant stages of tumor cells [19,32 –34]. In this study, we performed the first comparative evaluation of the sensitivity between piezoelectric (by QCM) and impedimetric (by EIS) approaches in determining lectin –cell interactions. In particular, the interactions between ArtinM lectin and NB4 leukemia cells were evaluated under Langmuir adsorption (or biochemistry-binding) assumptions. The NB4 leukemia cell line was chosen because of its known susceptibility to ArtinM death .
The ecotropic virus integration site 1 (EVI1) transcription factor is associated with human myeloid malignancy of poor prognosis and is overexpressed in 8–10% of adult AML and strikingly up to 27% of pediatric MLL-rearranged leukemias. For the first time, we report comprehensive genomewide EVI1 binding and whole transcriptome gene deregulation in leukemic cells using a combination of ChIP-Seq and RNA-Seq expression profiling. We found disruption of terminal myeloid differentiation and cell cycle regulation to be prominent in EVI-induced leukemogenesis. Specifically, we identified EVI1 directly binds to and downregulates the master myeloid differentiation gene Cebpe and several of its downstream gene targets critical for terminal myeloid differentiation. We also found EVI1 binds to and downregulates Serpinb2 as well as numerous genes involved in the Jak-Stat signaling pathway. Finally, we identified decreased expression of several ATP- dependent P2X purinoreceptors genes involved in apoptosis mechanisms. These findings provide a foundation for future study of potential therapeutic gene targets for EVI1-induced leukemia.
We then identified genes that were reported in multiple studies. Of the total 4,918 genes, 1,686 (34.3%) were reported in more than one study. We ranked genes that were listed in at least 8 studies by number of references, number of expression platforms, and number of expression features (Table 2). Although most of these genes have been associated with AML elsewhere in the literature, several genes (VCAN and PGDS) were only described in AML cell lines and a surprising number of the genes (HLA-DPA1, ITM2A, RBPMS, RGS10, RNASE2 and TRH) were not specifically described in AML. VCAN is a component of the extracellular matrix modulating cell adhesion, cell proliferation, cell migration, and extracellular matrix assembly. High expression of VCAN has been found in many malignancies, such as melanomas, ovarian, breast, and lung tumors, and in the acute monocytic leukemia cell line, THP-1. PGDS is an enzyme that catalyzes the conversion of PGH2 to PGD2, which is a prostaglandin involved in vasodilation, bronchoconstriction, inhibition of platelet aggregation, and recruitment of inflammatory cells. PGDS expression has been reported in two megakaryoblastic cell lines, CMK and Dami. TRH is a neurotransmitter/neuromodulator in