T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological malignancy resulting from the transformation of T-cell progenitors at various stages of development. They represent 10% of pediatric and 25% of adult ALL. Despite improvement in therapeutic protocols that cure nearly 80% children and 50% adults, relapse is common and the prognosis of relapsed T-ALL remains extremely poor [1, 2]. T-ALL oncogenesis results from chromosomal rearrangements, somatic genetic mutations, aberrant oncogene expression and impairment of multiple signaling pathways involving kinases [3–7]. Interestingly, the use of several kinase inhibitors has been shown to be
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ACAs: Additional cytogenetic abnormalities; ALL: Acute lymphoblastic leukemia; aMCB: Array-proven high-resolution multicolor banding; B-ALL: B cell acute lymphoblastic leukemia; BM: Bone marrow; Cdks: Cyclin-dependent kinases; CKS1: Cyclin kinase subunit 1; CNS: Central nervous system; DAPI: 4',6- diamidino-2-phenylindole; D-FISH: Dual-color fluorescence in situ hybridization; DNA: Deoxyribonucleic acid; EFS: Event-free survival; FISH: Fluorescence in situ hybridization; GMALL: German Multicenter Study Group for Adult Acute Lymphoblastic Leukemia; HLA-Dr: Human leukocyte antigen-antigen D related; ISCN: International System for Human Cytogenetic Nomenclature; LDH: Lactate dehydrogenase; PB: Peripheral blood; PBX1: Pre-B cell leukemia homeobox 1; RT-PCR: Reverse transcriptase-polymerase chain reaction; sIgM: Surface-bound IgM; T-ALL: T cell acute lymphoblastic leukemia; TCF3: Transcription factor 3; WBC: White blood cell; WCP: Whole chromosome paint; WHO: World Health Organization
Samples donated by patients with T cell acute lymphoblastic leukemia (T-ALL) were screened for mutations of the p53 tumor suppressor gene. Peripheral blood cells of T-ALL relapse patient H.A. were found to possess a heterozygous point mutation at codon 175 of the p53 gene. To determine whether this was an inherited mutation, a B cell line (HABL) was established. Leukemic T cell lines (HATL) were concurrently established by growing peripheral blood leukemic T cells at low oxygen tension in medium supplemented with IGF- I. Previously we had shown that > 60% of leukemic T cell lines possessed mutations in the p53 gene (Cheng, J., and M. Hass. 1990. Mol. Cell. Biol. 10:5502), mutations that might have originated with the donor's leukemic cells, or might have been induced during establishment of the cell lines. To answer whether establishment of the HATL lines was associated with the induction of p53 mutations, cDNAs of the HATL and HABL lines were sequenced. The HATL lines retained the same heterozygous p53 mutation that was present in the patient's leukemic cells. The HABL line lacked p53 mutations. Immunoprecipitation with specific anti-p53 antibodies showed that HATL cells produced p53 proteins of mutant and wild type immunophenotype, while the HABL line synthesized only wild-type p53 protein. The HATL cells had an abnormal karyotype, while the HABL cells possessed a […]
Early in vivo responses are known to be powerful pre- dictors of treatment outcome in childhood ALL [7 – 11]. Prednisone response (PR) was found to be related to treatment outcome by the Berlin-Frankfurt-Munster (BFM) study in 1983  and since then the predictive value of PR was confirmed in many studies [12–15]. T- ALL patients are more likely to have a worse steroid re- sponse than B-ALL patients [8, 10, 16]. Bone marrow morphology at day 15 during induction therapy is a well- established predictive factor and patients with ≥ 25 % blast cells in bone marrow usually have an inferior survival [9, 13, 17, 18]. During the last two decades, minimal re- sidual disease (MRD) in childhood ALL had been proved to be a remarkable predictive factor and already become an integral part of risk stratifications in many long established leukemia groups [19 – 24]. The most widely applicable MRD technique is polymerase chain reaction (PCR) analysis of T-cell receptor (TCR) and clone-specific immunoglobulin gene arrangements [25 – 27]. Although less standardized than molecular detection of MRD, flow cytometry (FCM) is faster, cheaper and more applicable [18, 22, 28 – 31]. Patients ’ prognosis and quality of life were further improved by individual- ized treatment. The majority of MRD studies were based on B-ALL, whereas MRD studies in T-ALL were scarce. A slower clearance of leukemia cells was found in T-ALL pa- tients and MRD risk group classified by MRD levels at the end of induction and before consolidation therapy was identified to be the most powerful independent prognostic factor in T-ALL patients [32, 33].
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T cell acute lymphoblastic leukemias (T-ALLs) are aggressive hema- tologic tumors resulting from the malignant transformation of T cell progenitors. T-ALL accounts for 10%–15% of pediatric and 25% of adult ALL cases (1) and is characteristically more frequent in males than females. Clinically, T-ALL patients show diffuse infil- tration of the bone marrow by immature T cell lymphoblasts, high white blood cell counts, mediastinal masses with pleural effusions, and frequent infiltration of the central nervous system at diagnosis. Although originally associated with high relapse rates, the progno- sis of T-ALL has gradually improved with the Introduction of inten- sified chemotherapy, with cure rates in modern protocols reaching over 75% in children and about 50% in adults with this disease (2). However, the outcome of T-ALL patients with primary resistant or relapsed leukemia remains poor (3, 4). Therefore, current research efforts are focused on the search for targets for the development of more effective and less toxic antileukemic drugs (5), which will likely require a greater degree of specificity and an improved under- standing of the molecular events that lead to the disease.
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recruitment of Treg cells [41–43]. An accumulation of sen- escent CD8 + CD28 − T cells was observed in several solid tu- mors, indicating the use of the suppressive activity of senescent T cells as a strategy for immune evasion [44–47]. Tumor-derived cAMP was shown to be responsible for the direct induction of senescence in T cells and is also a key component of the Treg cell mechanism of forcing T cells into senescence . These findings correlate with re-occurring observations of Treg cell accumulations in hematological malignancies such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), multiple myeloma (MM), and B cell lymphomas [49–52]. Conclusively, reduced Treg cell accumulation significantly prognosticated low relapse risk and leukemia-free survival (LFS) in AML patients [48, 49]. Recently, senescent T cells including clonally expanded CD8 + T cells with a CD28 − KLRG1 + CD57 + or CD28 − CD57 + PD-1 + phenotype were characterized in MM patients. Remarkably, these T cell clones showed telomere-independent senescence with upregulated tel- omerase activity indicating reversibility of senescence [50, 51]. Moreover, higher numbers of CD28 − CD57 + PD-1 + T cells were associated with early relapse in patients with MM after autologous stem cell transplantation (ASCT) . In addition, senescent and exhausted T cells in patients negatively affect T cell immunotherapy.
Another risk factor for ALL relapse was donor gender; in particular, male donors were associated with a higher RI. As male donors were associated with a lower inci- dence of grade II – IV acute GvHD (aGvHD, Additional file 4: Table S4), one possible reason for the higher inci- dence of relapse could be a lower GVL effect. Recently, McCurdy et al. described a higher PFS after PT-Cy Haplo-SCT if grade II aGvHD had occurred . Nakasone et al.  have previously reported lower inci- dence of relapse in a subset of male patients receiving allo-SCT from a female donor. Baron et al.  also re- ported a reduced incidence of acute leukemia relapse and a higher incidence of cGvHD in the combination of female donors to male patients. Of note, in our study, male do- nors were also a risk factor for a lower leukemia-free sur- vival (LFS) and OS but not for higher NRM both in ALL and in AML.
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The most evolutionary conserved arm of the UPR is me- diated by IRE-1α, which is activated by autophosphoryl- ation and oligomerization upon accumulation of misfolded proteins in the ER. IRE-1α contains an endoribonuclease (RNase) and a kinase domain. So far, the most described function of the RNase domain is to reduce ER load through unconventional splicing of the X-box binding protein 1 (XBP1) mRNA . This unconventional splicing leads to removal of a 26-nucleotide intron. Recently, it was uncov- ered that a multimeric protein complex tRNA splicing lig- ase is responsible for ligation of IRE-1α-cleaved XBP1 mRNA 5′ and 3′ extremities of which the RNA 2′,3′-cyclic phosphate and 5′-OH ligase (RTCB, also named HSPC117/ C22orf28) seems to be the most essential subunit [19–22] and demonstrated a physiological role in plasma-cell differ- entiation . This causes a frameshift in the XBP1 reading frame, thus generating a transcriptionally active protein (XBP1s). XBP1s controls expression of genes involved in protein folding, secretion, ERAD, and lipid synthe- sis [23, 24]. IRE-1α RNase activity is also involved in
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Bacterial or fungal infections associated with neutropenia are common in patients with newly diagnosed or recurrent acute leukemia (32, 33). Three of our five patients with CNS infections had persistent neutropenia. One patient (case 9) with purulent meningitis had enhancement of the leptomeninges of the basal surface of the brain and spinal cord on postcontrast T1- weighted MR images (Fig 6). A follow-up CT scan before death showed multiple infarcts re- sulting from infection-induced vasculitis, a find- ing verified at autopsy. Among the three pa- tients with fungal infections, one (case 13) had pathologically proved cerebral mucormycosis. Isolated cerebral mucormycosis is uncommon and usually results from extension of preexist- ing infection by a species of Mucor in the nose or paranasal sinuses (rhinocerebral mucormy- cosis) (34, 35). A preoperative imaging diag- nosis of cerebral mucormycosis is difficult. The hypointense signal intensity of the mucormyco- sis seen on T2-weighted images (Fig 15B) is not understood, although a recent study sug- gested that the low signal intensity of Aspergil- lus fumigatus on T2-weighted and gradient- echo MR images was caused by accrual of the paramagnetic substances, such as iron, man- ganese, magnesium, and zinc (36). Microbio- logical documentation of CNS fungal infection (cases 11 and 12) was not obtained, despite cultures from urine, blood, and CSF. In case 11,
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count of 156 × 10 9 /l. Biochemistry analyses revealed serum lactate dehydrogenase (LDH) value was 893.2 U/l (normal value up to 480 U/l); serum aspartate aminotrasferase (AST) level was 42 U/l (normal up to 45 U/l); and alanine aminotransferase (ALT) level was 122 U/l (normal up to 45 U/l). Total serum protein was within normal range at 7.1 gm/dl (normal value 6.4–8.3 gm/dl) but serum albumin was 4.2 gm/dl (normal value 3.2–5 gm/dl). Ferritin value was 1349 (13–150 ng/ml). Bone marrow aspiration revealed 95% of blasts. At this point the first cytogenetic and immu- nophenotypic data were determined; simultaneously she had been diagnosed as having B-ALL based on clinical and pathological data. Treatment with hyper-CVAD for overall 10 months was initiated. The patient did not respond to that treatment and suffered from hematuria, right eye vision deterioration, gastrointestinal bleeding, and fever; she received blood transfusion many times, and her periph- eral blood (PB) showed pancytopenia. One month after initiation of hyper-CVAD treatment her PB reached a short amelioration of different cell counts (WBC, plt, RBC) together with severe diarrhea, fatigue and weakness. One week later the patient showed pancytopenia again with diarrhea, severe stomach heart burn and fever. After one month the BM smear showed less than 40% blats cells with hypocellularity and BM suppression. She received cortisone, and two months later BM revealed again / still pancyto- penia, suppression and myelofibrosis. After another two months BM regenerated and WBC count was 66 × 10 9 /l (90% were blasts). Nonetheless, the patient succumbed due to unknown causes whilst under treatment. Her cousin agreed with scientific evaluation of her case and the study was approved by the ethical committee of the Atomic En- ergy Commission, Damascus, Syria.
Acute myeloid leukemia (AML) is a group of clinically and genetically heterogeneous diseases [1,2]. Despite treat- ment advancements in acute promyelocytic leukemia (M3), current treatment of AML is based on chemother- apy. Standard induction chemotherapy consists of anthra- cycline and cytarabine (3 + 7 regimen) can achieve the complete remission (CR) rate of approximately 75%, but outcome is uncertain because of the variability of individ- ual genetic profile and drug sensitivity [3,4]. Intense chemotherapy or allogeneic hematopoietic cell transplant- ation (allo-HSCT) can benefit patients who are refractory or tend to relapse . Early and easy monitoring of min- imal residual disease (MRD) reflects treatment response in time and becomes an essential reference for patients with AML to optimize chemotherapy.
Metastases to the breast and axilla are rare and account for approximately 2% of all mammary malignancies [1,2]. The most common metastatic lesion to involve the breast is a metastasis from a contralateral mammary cancer [1,2]. If hematologic malignancies are also excluded, the number of non-mammary metastases drops to well below 1% [1-3]. Although most common extra mammarian metastases of breast is from hematopoetic system and melanomas, presenting of the patient with both breast involvement is very rare . The present study reports the rare case of thirty-year-old female breast metastases with extra mammarian leukemia and lymphoma. Due to the rarity of the disease, the relevant literature was also reviewed.
Various techniques have been used in the literature to improve the diagnostic process of non-healthy cells in the blood smear images. For instance, Rawat, J et al. (2015)  offered a system that can differentiate between healthy and non- healthy cells in an ALL-IDB dataset by using the shape and texture features as the input to the SVM classifier with achieving an accuracy of 89.8%. When Dumyan, S. and Gupta, A. (2017)  used Artificial Neural Network (ANN) classifier depend on the shape, texture, statistical and moment invariant features extracted from 36 blood smear images to identify ALL cells, the overall accuracy increased to 97.9%. Negm, A. et al. (2017)  proved that ANN and Decision tree classifier successfully discriminated the leukemia cells by using geometry, color and relative tissue features extracted from 642 images which are collected from a local hospital with achieved an accuracy of 99.519%. However, the study did not distinguish among the various FAB subtypes.
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Patients were not enrolled in any clinical trials, but received treatments either according to the Brazilian Group for Treatment of Childhood Lymphoblastic Leukemia (GBTLI-ALL 99/09) or to ALL Berlin- Frankfurt- Münster (BFM-ALL 95 and 2002) protocols. 17,18 Patients were assigned into the group of a high risk of relapse, according to age strata, leukocyte count (>50×10 9 /L), and T-cell phenotype of both proto- cols. In summary, all patients received a 4-week induc- tion phase, which included prednisone, vincristine, doxorubicin, L-asparaginase, intrathecal Methotrexate (MTX), Cytarabine (Ara-C), and dexamethasone (MADIT). In the GBTLI-ALL, the consolidation phase, Cyclophosphamide, Ara-C, and 6-mercaptopurine (6-MP) were included. Intensi ﬁ cation treatment with MTX 2 g/m 2 ×4, 6-MP and MADIT and maintenance of 6-MP, MTX, vincristine, dexamethasone, and MADIT. Patients with more than 5% blast cells in their cere- brospinal ﬂ uid after day 14 of induction therapy have received cranial radiotherapy (1200 Gy during the late consolidation phase). 18
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PF-0477736 is a selective and competitive inhibitor for the CHK1 ATP site. Its specificity is 100 times stronger for CHK1 than that for CHK2. The efficacy of this com- pound has been well established against different kinds of tumor. In ovarian cancer, it has been shown that tumor cells strongly respond to treatment with PF- 0477736 but they generate metastasis and chemo- resistant clones . The efficacy of PF-0477736 has been evaluated also in leukemia. Sarmento et al.  demonstrated that the T-ALL primary samples express higher level of CHK1 kinase in comparison to normal thymocytes. The treatment with PF-0477736 promoted apoptotic cell death and CHK1 inhibition and conse- quently impaired replication and abrogation of G2/M checkpoint in T-ALL cells. Interestingly, in vitro treat- ment did not significantly affect the viability of normal thymocyte cells . Similar results have been shown by our group. The inhibition of CHK1/CHK2 by PF- 0477736 as single agent deeply reduced the cell viability of ALL primary cells and leukemia cell lines. The results from the in vitro/ex vivo studies were further confirmed using an in vivo model . Recently, Nguyen T. and colleagues reported the in vitro/in vivo synergic efficacy of PF-00477736 in combination with the Src/ABL inhibi- tor bosutinib (SKI-606) in BCR-ABL1-positive CML or ALL cells, focusing on highly imatinib-resistant models with ABL kinase mutations. The authors speculated that the combination acts through a BCR-ABL1-independent process that may involve multiple mechanisms, includ- ing inactivation of ERK1/2 and Src, up-regulation of BIM, down-regulation of MCL-1 (BCL-2-like protein), activation of CDK1, and induction of DNA damage .
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feasibility of monitoring the cytotoxic effects of several che- motherapeutic agents on different leukemia/lymphoma cells (Figure 4). We include in Figure 4B the percentage values for each dose with respect to the control (non-treated) for both the RTCA and the MTT method. The percentages obtained from the two methods were comparable. Only at certain non-cytotoxic concentrations (1 nM trabectedin L1236 or 1 nM trabectedin/doxorubicin, Jurkat) we observed significant increases in the CI that did not correspond to significant increases in the MTT values. We attribute this to the fact that, at such low concentrations, the activity of the mitochondrial succinate dehydrogenase is not affected; nevertheless, certain cell morphology parameters are modified, and thus may be monitored, revealing additional data. Finally, we also show that under these experimental conditions, the MTT values obtained when cells are incubated in the absence/presence of fibronectin were similar (Figure 4B). Although not sig- nificant, we observed a slight tendency to survive in cells incubated in the presence of fibronectin.
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Here we present a male patient with acute myeloid leukemia (AML) initially diagnosed as M5 and with karyotype 46,XY. After induction therapy, he underwent a HLA-matched allogeneic hematopoietic stem cell transplantation, and six years later he relapsed as AML M1 with an abnormal karyotype //47,XX,+10/47,XX,+11/48,XX,+10,+11 /46,XX. Based on this, we tested the possibility of donor cell origin by FISH and molecular STR analysis. We found no evidence of Y chromosome presence by FISH and STR analysis consistent with the success of the allogeneic hematopoietic stem cell transplantation from the female donor. FISH studies confirmed trisomies and no evidence of MLL translocation either p53 or ATM deletion. Additionally 28 fusion common leukemia transcripts were evaluated by multiplex reverse transcriptase-polymerase chain reaction assay and were not rearranged. STR analysis showed a complete donor chimerism. Thus, donor cell leukemia (DCL) was concluded, being essential the use of cytological and molecular approaches. Pediatric DCL is uncommon, our patient seems to be the sixth case and additionally it presented a late donor cell leukemia appearance. Different extrinsic and intrinsic mechanisms have been considered to explain this uncommon finding as well as the implications to the patient.
mononuclear cells from healthy donors were not significantly affected by idelalisib. In order to mimic the bone marrow microenvironment, which is thought to be protective for malignant cells in plasma cell myeloma, bone marrow stem cells, interleukin-6, and insulin-like growth factor-1 were added to plasma cell myeloma cells in culture but these did not provide protection against idelalisib-induced apoptosis and cell death. The authors also hypothesized that idelalisib would have antiangiogenesis activity; the drug was tested on human umbilical vein endothelial cells to assess for inhibi- tion of capillary-like tube formation. Idelalisib inhibited capillary-like tube formation in a dose-independent and time-dependent manner. 14 Additionally, idelalisib showed a
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Given the limitation in child hood cancer patient numbers that severely restrict adequate early phase trials using genetically modified T cell receptors, it is anticipated that most information would arrive from research undertaken in the domain of adult oncology, or in trials that comprise of both adult and paediatric patients. Ultimately, however, proper and well designed clinical trials would any ways be required to be conducted in children to estimate the efficacy and toxicity of this new modified T cell therapy. Assuming that suitable and desired trial results emerge, we anticipate dramatic surge in efforts for large scale manufacturing of these molecules at commercial scale to reduce cost and make the therapy cost effective.
marrow smear analysis revealed an abnormal increase in the amount of lymphoblast and Immature lymphocytes (60% and 88.5%, respectively) (Figure 3A), which indicated ALL (L2). Flow cytometry showed abnormal B cells (68.6%) (Figure 3B-F). Fluorescence in situ hybridization (FISH) indicated mixed lineage leukemia (MLL) fusion gene was positive. Addi- tionally, Ig and TCR Gene rearrangement detec- tion were negative. Chromosome-based ana- lysis showed structural rearrangements were also negative. According to World Health Orga- nization (WHO)-based classification, these re- sults verified the diagnosis as B-ALL (L2, BII, MLL-ENL positive, high-risk). Regular chemothe- rapy was began with chemotherapy regimen of high-risk B-ALL according to the modified Figure 1. Histopathological images of case 1. A: The left renal and tumor by resection, hematoxylin-eosinstain; origi- nal magnification ×200. B: Hematoxylin-eosin stain; original magnification ×400. A broad range of spindle cells were in a woven arrangement. Part of the cell cytoplasm was with rich red dye. Cell atypia and deeply stained nuclear were significantly observed. C: Desmin (one of immunohistochemistry) of spindle cells was remarkably positive.
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