bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Novel Type of ETV6-TTL Fusion Gene in Pediatric Myeloid/NK Cell Precursor Acute Leukemia Having t(12;13)(p13;q14).
Authors: Taki Tomohiko , Hayashi Yasuhide , Taketani Takeshi , Sako Masahiro , Taniwaki Masafumi 
Published: October 2019

Abstract
The t(12;13)(p11-13:q11-14) is a recurring chromosomal abnormality found in different types of hematological malignancies. Three genes on 13q12-14, CDX2, FLT3, and TTL have been identified as fusion partners of ETV6(TEL) on 12p13. These genes create different forms of fusion transcripts with ETV6. In the present study, we identified novel types of ETV6-TTL fusion transcripts in a case of a 9 year-old female who developed myeloid/NK cell precursor acute leukemia with t(12;13)(p13;q14). Cytogenetic analyses of the leukemic cells of the patient using a regular G-banding revealed 47, XX, del(2)(q?), -9, der(12)t(12;13)(p13;q14), add(13)(q1?2), +mar1, +mar2 [13/20]. Fluorescence in situ hybridization using YAC clone revealed that ETV6 gene was split in metaphase chromosomes of patient’s leukemic cells. We performed reverse-transcriptase-polymerase chain reaction (RT-PCR) analysis to detect the fusion transcripts of ETV6-CDX2, ETV6-FLT3 or TTL-ETV6, however, no fusion transcripts of previously described types were detected. We next performed RT-PCR analysis using various sets of primers to detect unknown types of fusion transcripts involving these genes, and detected novel types of fusion transcripts of ETV6-TTL. These fusion transcripts consisted of exons 1 to 5 of ETV6 and exons 5 to 8 of TTL, and exons 1 to 6 of ETV6 and exon 9 or exons 8 to 9 of TTL. No reciprocal fusion transcripts were detected. Predicted fusion proteins consisted of N-terminal ETV6 lacking whole or part of ETS-binding domain and C-terminal TTL. Previous report showed major type of TTL-ETV6 fusion transcript consisted of exons 1 to 5 of TTL and exons 2 to 8 of ETV6 which contained both helix-loop-helix and ETS-binding domains. These results suggested that novel types of ETV6-TTL act as different fusion proteins from previously reported TTL-ETV6 in leukemogenesis. At the cytogenetic level, it may be difficult to distinguish ETV6-CDX2, ETV6-FLT3, and ETV6-TTL. Further accumulation of the patients with t(12;13) and further analysis of these novel types of fusion transcripts may clarify the pathogenesis of t(12;13)-leukemia.
Acute Myeloid Leukemia (AML-M2) with a Novel Translocation t(5;11)(q35;q13) and Normal Expression of Cyclin D1.
Authors: de Oliveira Fabio M. , dos Santos Guilherme A.S. , Marinato Andre F. , Araujo Amelia G. , Lima Ana S.G. , Jacomo Rafael H. , Falcao Roberto P. , Tone Luis G. , Simoes Belinda P. , Rego Eduardo M. 
Published: October 2019

Abstract
Rearrangements of the long arm of chromosome 11 are commonly associated with acute leukemias, with the breakpoints clustered mainly to the 11q23 region, frequently leading to the rearrangement of the MLL gene. CCND1 (previously PRAD1, BCL1), the gene encoding cyclin D1, is located at 11q13. Overexpression of cyclin D1 represents one of the common genetic alterations in human neoplasia, leading to a change in G1-S transition and uncontrolled cell growth. Here we describe the case of a patient with acute myelogenous leukemia (AML) harboring a chromosomal translocation involving 11q13 and a new partner, chromosome 5. A 30-year-old male presented with dyspnea for one month and peripheral blood counts of: hemoglobin 4.9 g/dL, platelets 62 x 109/L and leukocytes 29.5 x 109/L (with 45% of blasts). Bone marrow examination showed a hypercellular marrow with 82% blasts positive for myeloperoxidase and negative for non-specific esterase. The case was classified as AML M2 according to FAB. Classical cytogenetics and spectral karyotyping (SKY) studies performed by unsynchronized culture of the marrow cells revealed an abnormal clone of 46,XY, t(5;11)(q35;q13)[20]. RT-PCR for the rearrangements MLL/AF9, MLL/AF6, MLL/AF4, MLL/ENL, and MLL/ELL were performed and were negative for all of them. Cyclin D1 overexpression was not detected in bone marrow cells by Real Time PCR. The patient was submitted to induction chemotherapy with Daunorrubicin and Cytarabine but obtained only partial remission after 2 cycles of chemotherapy (10% of blasts in bone marrow after second induction). He was than submitted to an allogeneic stem cell transplantation (SCT) from his HLA identical sister. On day +30 after SCT he was in complete hematological remission. The classical cytogenetics study after chemotherapy and bone marrow transplantation revealed karyotypes 46,XY[20] and 46,XX[15]/46,XY[15]chi, respectively. The t(5;11)(q35;q13) represents a recurrent abnormality in renal oncocytoma (benign tumors that occur predominantly in the kidney) and leads frequently to cyclin D1 overexpression. Nevertheless, this cromossomal translocation has never been described in AML.
Quantitative Analysis of Wilms’ Tumor Gene 1(WT1) Expression in Acute Lymphoblastic Leukemia (ALL) of Adults Reveals Lower WT1 Levels Than in Acute Myeloid Leukemias (AML) and Varying Expression Levels between Individual Patients among Different ALL Subgroups.
Authors: Siehl Jan M. , Thiel Eckhard , Hoelzer Dieter , Schwartz Stefan , Keilholz Ulrich 
Published: October 2019

Abstract
Background: Overexpression of the embryonic transcription factor WT1 is common in human acute leukemias. Therefore WT1 became an interesting target mainly for immunotherapeutic approaches in AML. In previously published data from qualitative PCR analysis the frequency of WT1 overexpression in ALL is lower than in AML. But so far only limited data from quantitative analysis of WT1 expression in ALL are available.
Methods: In the present study we analysed bone marrow or blood samples of 238 adult ALL patients at diagnosis for the expression levels of WT1 by real-time RT - PCR (LightCycler). All patients were treated in the German Multicenter Study for the Treatment of ALL (GMALL 07/2003). Only samples containing at least 60 % blasts were used for analysis. Expression levels were compared to previously published data of WT1 expression levels in AML samples.
Results: WT1 expression was found in 219 out of 238 ALL samples (92 %). Compared to AML WT1 expression levels in ALL were significantly lower. There were no differences between the various ALL immuno phenotypes of the B or T lineage, but lymphoblasts with a coexpression of myeloid markers had significantly higher WT1 expression levels than those without a coexpression of myeloid markers. Surprisingly, patients who were bcr-abl negative had significantly higher WT1 expression levels than bcr-abl positive patients.
Conclusions: Using quantitative RT-PCR technology WT1 expression can be found in most ALL cases. There are usually less intense expression patterns compared to AML and expression varies among different ALL subgroups. Thus WT1 has promise to be an interesting target also in ALL therapy but in a more individually determined fashion. The relation of WT1 expression level and treatment outcome is currently being analysed.
Adult Acute Lymphoblastic Leukemia and Cytogenetic Abnormalities in Different Racial and Ethnic Subgroups.
Authors: Tamkus Deimante , Jajeh Ahmad , Berko Ebenezer , Osafo David , Griza Decebal , Kovarik Paula , Catchatourian Rose 
Published: October 2019

Abstract
Inferior survival of African American and Hispanic patients with acute lymphoblastic leukemia (ALL) has been reported. The reason for this is unclear. A retrospective analysis was conducted to see if abnormal cytogenetics account for the differences. We have analyzed cytogenetic studies of 39 adults (16 year and older) with newly diagnosed ALL who were consecutively treated at John Stroger Hospital of Cook County between 1997 and 2005. The study population included 13 (33%) African Americans, 18 (46%) Hispanics, 6 (16%) Caucasians, and 2 (5 %) other ethnic background adults. Male to female ratio was 2:1. Mean age at diagnosis was 26 (range between 16 and 71 years). A clonal cytogenetic abnormality was detected in 27 patients, and 12 patients had normal karyotype. Patients with t (9;22)(q34;q11), BCR-ABL + by FISH, t (4;11)(q21;q23), +8, −7 cytogenetic abnormalities were assigned to an unfavorable cytogenetic group. This group was composed of 14 patients. None of our study patients had favorable cytogenetics: del (12p), t(12p), t(14q11-q23), inv(14)(q11;q32) or t(10;14)(q24;q11). Remaining 25 patients with normal karyotype (n=12) or miscellaneous cytogenetic abnormalities (n=13) were classified as normal risk group. 54 % of African Americans had unfavorable cytogenetics, compared with 33 % Caucasians and 17 % Hispanics. Translocation t (9;22) alone or in association with other cytogenetic abnormalities was most commonly seen. 9 patients had single, and 18 had multiple chromosomal changes. Hispanic group had more complex cytogenetic changes when compared with the African American or Caucasian groups. Unfavorable cytogenetic abnormalities may account for inferior survival in African Americans, but other factors such as compliance or pharmacogenetics should be evaluated, especially in the Hispanic patient population.
The Wnt/β-Catenin Pathway Promotes Growth and Survival of Acute Lymphoblastic Leukaemia (ALL) Cells In Vitro.
Authors: Khan Naveed I. , Bradstock Kenneth F. , Bendall Linda J. 
Published: October 2019

Abstract
Wnt signalling is known to play an important regulatory role in self-renewal of haematopoietic stem cells as well as development of normal lymphocyte progenitors. Recently this pathway was implicated in a number of haematological malignancies including AML, CML, MM and CLL. However its role in acute lymphoblastic leukaemia is not clear. We have previously shown that primary pre-B ALL cells and cell lines express various Wnt family members, receptors and downstream transcription factors. We extended these studies further to elucidate a functional role of Wnt signalling in leukemic cells. Exposure to Wnt3a, a ligand inducing the canonical Wnt signal cascade, resulted in stabilisation of beta-catenin and its nuclear translocation in all growth factor independent pre-B cell lines (n=3), stromal-dependent cell line (n=1) and primary cases (n=3) examined. Wnt3a treated pre-B ALL cell lines and a patient sample were more resistant to apoptosis induced by serum deprivation (viability on day 4, 51.3±18.4% Wnt3a treated vs 29.5±19.9% control treated, p-value <0.05). 3H-thymidine incorporation studies revealed 4.0±1.7 fold increase in proliferation which was associated with increased cell cycle entry as determined by Ki-67 staining. Cell cycle analysis confirmed an increase in the proportion of cells in S and G2M phases of cell cycle. These effects were associated with increased transcriptional activity of toposiomerase II-alpha as assessed by semi-quantitative PCR and protein expression. Taken together our results indicate that Wnt-dependant β-catenin signalling pathway is active in pre-B ALL cells and promotes their growth and survival in vitro.
Epidemiology of Adult Acute Lymphoblastic Leukemia in Manitoba, Canada: Does This Approximate Clinical Trial Data?.
Authors: Dao Vi , Rubinger Morel , Bow Eric J. , Schacter Brent A. , Shore Tsiporah B. , Woloschuk Donna , Demers Alain , Sun Xiaoyun , Seftel Matthew D. 
Published: October 2019

Abstract
Background: Outcomes of unselected adult ALL patients (pts) have not been well studied at a population level. Whether such pts behave differently to those in formal clinical trials is unclear. We developed a uniform therapeutic protocol as the provincial standard of care in Manitoba, a province with 1.2 million people, for adult pts since 1989 (“ALL89-1”). The aim of this retrospective study was: to ascertain the local epidemiology of adult ALL; to analyze the efficacy and toxicity of the ALL89-1 regimen and any other regimens given to pts in a population-based setting.
Methods: All pts aged ≥18 years (yrs) diagnosed with ALL in Manitoba between 01/1989 and 11/2004 were identified via the provincial cancer registry. A review of computerized and paper charts was conducted. We focused on demographics, diagnostic clinical characteristics, therapy, treatment related toxicities, and whether stem cell transplantation (SCT) was performed.
Results: There were 81 pts with a median age of 44yrs (range, 18–90). 53 (65%) pts were male. 58 pts received ALL89-1, 12 received other regimens and 11 patients received palliation only. Thirteen pts were Philadelphia chromosome positive (Ph+). Complete remission (CR) rate after one cycle of induction therapy was 74% for all pts, and 74% for ALL89-1 pts. At a median follow-up of 56 months (range 7–265), estimated overall survival (OS) and leukemia free survival (LFS) at 5 years was 26% (95% Confidence interval [C.I.], 15–36) and 24%( 95% C.I. 14–35). The 5 year OS and LFS for the ALL89–1 group was 23% (95% C.I. 11–35) and 20% (95% C.I. 8–32). There were 13 treatment related deaths in the 58 ALL 89-1 pts, 4 of whom died post SCT. In multivariate analysis for all 81 pts, attainment of CR post induction chemotherapy independently predicted improved survival (hazard ratio 0.23, 95% C.I. 0.11–0.5). In contrast, age > 55yrs, Ph+ status, and high WBC at diagnosis were not significant survival predictors. Palliative pts survived for a median of 10 days (range, 0–53) from diagnosis.
Conclusions: This comprehensive province-wide study of adult ALL pts treated at a single cancer centre shows that durable leukemia free survival is possible, but for only a small subgroup. These results appear inferior to results of published clinical trials. The reasons for the relatively poor outcomes require further analysis in this and other jurisdictions.
Myeloid Blastic Cells with MLL-ELL Translocation Can Convert Their Morphology into the Fibroblastoid Cells.
Authors: Tashiro Haruko , Noguchi Mitsuho , Shirasaki Ryosuke , Goto Moritaka , Kawasugi Kazuo , Shirafuji Naoki 
Published: October 2019

Abstract
Objective: It has been reported that acute leukemia with the fusion product of MLL-related molecule is determined to be the poor prognosis. To make clear one of its biological characters, we cultured AML blasts with t (11; 19) (q23; p13.1), which produced fusion protein of MLL and ELL, and the morphological changes as well as their DNA, and RNA were examined.
Method: Bone marrow cells were obtained from two patients with MLL-ELL fusion gene after obtaining informed consent. There were over 90% blastic cells in bone marrow. Mononuclear cells were obtained with Ficoll (SG. 1077), and were cultured in DMEM with 10% FCS in CO2 incubator. The morphology of the cells was observed for 3 months. Also, DNA was extracted, and MLL-ELL translocation was analyzed with genomic Southern blotting with probe x which contains BamHI-digested 0.9 kb fragment from MLL cDNA. RNA was extracted with GTC method, and 1st strand cDNA was synthesized with oligo-dT primer, and PCR was performed with MLL common primer and ELL antisense primer. The reaction of DNA sequencing was performed with BigDye Terminator Cycle Sequencing kit (Applied Biosystems), and analyzed with ABI Prism 3700 DNA analyzer.
Result: After one month of the cultures, fibroblastoid cells were expanded without any blastic cells onto them. Southern analysis, RT-PCR study, and DNA sequence revealed that the fusion cDNA product with MLL and ELL was observed in fibroblastoid cells.
Discussion: AML blasts with MLL-ELL fusion gene can convert their morphology into the fibroblastoid cells. The fibroblasts have been reported to be resistant to the chemotherapeutic agents. This morphological conversion may contribute one of the causes of poor prognosis in AML patients with MLL-ELL fusion gene. We now characterize precisely these fibroblastoid cells in point of the expression of the specific proteins, and the functions including the binding and proliferating capability to the non-adherent blastic cells.
Platelet-Derived Factor VIII (FVIII) Corrects the Murine Hemophilia A Phenotype Even in the Presence of FVIII Inhibitors.
Authors: Shi Qizhen , Wilcox David A. , Fahs Scot A. , Cooley Brian C. , Desai Drashti , Weiler Harmut , Morateck Patricia A. , Montgomery Robert R. 
Published: October 2019

Abstract
Inhibitory antibody development is not only the most severe and important complication of FVIII infusion in hemophilia A patients, but also a major potential concern in patients following hemophilia A gene therapy. We have developed a transgenic model in which human B-domain deleted FVIII (hFVIII) expression is controlled by the platelet-specific αIIb promoter (2bF8) and results in FVIII storage together with VWF in platelets. The transgenic mice were then bred into FVIII knockout mice (FVIIInull) so that the only FVIII in peripheral blood is derived in platelets. The bleeding phenotype was corrected in the 2bF8/FVIIInull transgenic mice, even though FVIII was not detectable in plasma. We further hypothesize that platelet-derived FVIII in a complex with VWF will 1) protect FVIII from inactivation by circulating FVIII-specific antibodies and 2) release FVIII in sufficiently high local concentration at sites of vessel damage so as to overcome immediate inactivation by FVIII inhibitors. To address our hypothesis, we used three strategies -1) an acute model infusing plasma from highly immunized FVIIInull mice, 2) immunization of 2bF8 transgenic mice with human FVIII, and 3) a model using transplantation of spleen cells from highly immunized mice into sub-lethally irradiated 2bF8 transgenic mice. Thirteen mice were infused with plasma containing 48-64 BU and 12 survived following tail clipping using a 1.59 mm template. FVIII-immunized transgenic mice (given adjuvant) generated anti-FVIII inhibitory antibodies as high as 350 BU after a single immunization. The level of platelet-FVIII and the platelet number in these mice were not significantly different from pre-immunization levels and 13 of 15 survived tail clipping. In the 3rd group, spleen cell transplantation resulted it titers of >5000 BU and 8 of 12 transgenic mice survived tail clip. In contrast, only 2 of 7 wild type mice survived tail clip with inhibitor titers >5000 BU after spleen cell transplantation. None of 9 untransplanted FVIIInull mice survived. These results demonstrate that platelet-derived FVIII is protected from inhibitor inactivation and correct the phenotype of murine hemophilia A even in the presence of a high-titer anti-FVIII antibody. This approach may be useful not only for preventing inhibitor formation in hemophilia A gene therapy but also for gene therapy in the case of pre-existing inhibitors.
Investigation into the Pro-Apoptotic and Pro-Inflammatory Properties of 6-Thioguanine and 6-Mercaptopurine.
Authors: Bruneau Johanna C. , Jenkins P. Vincent , O’Marcaigh Aengus , Smith Owen P. 
Published: October 2019

Abstract
The thiopurines, 6-mercaptopurine (6MP) and its analog 6-thioguanine (6TG), have considerable anti-leukaemic activity. 6MP has contributed to the cure of tens of thousands of children with acute lymphoblastic leukaemia (ALL) with little in the way of significant toxicity and hence it is now established as the purine nucleoside of choice in maintenance therapy of childhood ALL. This is despite the fact that 6TG may indeed be a better alternative to 6MP as it has been shown to be at least one log more active in vitro than 6MP and in a recent large U.S. ALL Trial, isotoxic comparisons of 6MP and 6TG showed 6TG to have fewer CNS events. However, like in a similar recent U.K. study there was increased toxicity noted in this U.S. Trial, namely the emergence of an inflammatory response-like syndrome affecting the liver resulting in sinusoidal occlusion (SOS) with hepatomegaly, thrombocytopenia, ascites, and elevated bilirubin and liver transaminases. Although these purine nucleosides have been in widespread use for more than 50 years, their exact cytotoxic mechanism of action remains to be elucidated with incorporation of the nucleoside analogs into DNA, resulting in apoptosis, and by the inhibition of de novo purine synthesis leading to metabolic arrest being the two main contenders. The mechanism of the inflammatory insult causing SOS with 6TG is also poorly understood. In order to identify the differential effects of 6-TG and 6-MP in vitro, we measured apoptosis via the TUNEL assay, viability using the WST-1 assay, and the secretion of inflammatory cytokines (TNF-a and IL-8) by ELISA in a panel of human leukaemia cell lines: THP-1 (acute monocytic leukaemia), Jurkat E6.1 (acute T lymphocytic leukaemia), 697 (acute pre-B lymphocytic leukaemia), as well as the hepatocyte cell line HepG2 after treatment with either of the two drugs. There was a significant (p≤ 0.01) increase in apoptosis in THP-1and 697 cells following treatment with 6-TG but not 6-MP, although there was a significant decrease in viability observed with both agents (p≤ 0.01). In contrast, a significant (p≤ 0.01) increase in apoptosis was observed in Jurkat E6.1 cells after treatment with both 6-TG and 6-MP. Neither apoptosis nor viability was significantly affected by either 6-TG or 6-MP in HepG2 cells. IL-8 secretion was significantly (p≤0.01) increased by 6-TG and 6-MP in THP-1 and HepG2 cells, however there was no corresponding increase in TNF-a levels. Neither agent had a significant effect on the secretion of either TNF-a or IL-8 in 697 or Jurkat E6.1 cells. These results show that there are different apoptotic and pro-inflammatory responses to these drugs most likely reflecting their efficacy and toxicity profiles
Amphotericin B and Itraconazole as Empirical Antifungal Therapy in Children with Acute Leukemia with Neutropenic Fever.
Authors: Kim Sun-Young , Park Jong-Sun , Jeong Nak-Gyun , Jeong Dae-Chul , Cho Bin , Kim Hack-Ki 
Published: October 2019

Abstract
Purpose: Fungal infections are one of the important causes of morbidity and mortality in patients with hematologic malignancies. Amphotericin B (ABV) and itraconazole (ITZA) have been used as the standard empirical antifungal therapy in neutropenic patients with acute leukemia who have persistent fever that does not respond to antibiotic therapy. ABV is an antifungal drug associated with side effects such as fever and chills, symptoms which may be mediated by pro-inflammatory cytokines such as interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). We assessed modulation of these pro-inflammatory cytokines as well as the anti-inflammatory cytokines (IL-4, IL-1Ra) by ABV and ITZA.
Methods: From October 2004 to February 2005, a total of 30 episodes from acute leukemia patients with febrile neutropenia were analyzed for this study. They were randomly allocated to receive intravenous ABV or ITZA for 14 days. Clinical responses were evaluated at the completion of therapy, and cytokine IL-1β, TNF-α, IL-4, and IL-1Ra were measured for determination to know the correlation between two antifungal agents and inflammatory cytokines.
Results: Empirical antifungal agents were given to 37 patients (ABV 20, ITZA 17), and 30 patients (ABV 15, ITZA 15) were evaluable for efficacy. White blood cell and absolute neutrophil count in the group treated with ITZA increased early days of treatment, so the duration of neutropenia in ITZA group is shorter. Serum creatinine level is lower in ITZA group than in ABV group but this is not statistically significant. There was no significant difference in response rate between two groups. The IL-1β was increased in ABV treatment group and the ratio of IL-1Ra/IL-1β is markedly decreased in ABV treatment group while increased in ITZA group.
Conclusions: ITZA and ABV have at least equivalent efficacy as empirical antifungal therapy in neutropenic children with acute leukemia. However ITZA is associated with significantly less toxicity in clinical and molecular aspects.