bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bcr-Abl and Wilms Tumor Gene (WT1) Kinetics in Nilotinib (AMN107) Treated CML Patients.
Authors: Schwarz Michaela , Baskaynak Gokben , Turkmen Seval , Schuld Peter , Dorken Bernd , Daniel Peter , Philipp le Coutre 
Published: October 2019

Abstract
Background: In normal hemopoesis WT1 expression is restricted to early progenitors and is rapidly down-regulated in maturing blood cells. WT1 transcript quantification therefore may be an adjunct diagnostic method in CML patients (pts) that remain Bcr-Abl positive under treatment. Especially, in advanced CML pts WT1 transcript kinetics may precede a change in Bcr-Abl kinetics and may therefore be helpful to indicate early response to treatment. Nilotinib is a potent, highly selective, second generation Bcr-Abl inhibitor which in vitro is 30-fold more potent than imatinib and is currently studied in imatinib resistant or intolerant pts.
Aim: The aim of this study was to compare kinetics of Bcr-Abl and WT1 transcripts in 33 CML pts with late chronic phase (CP) or advanced disease under treatment with nilotinib.
Methods: All pts were treated as part of an ongoing phaseI/II clinical trial with nilotinib at a standard dose of 400 mg BID. 20 pts were in chronic phase (CP), 4 in accelerated phase (AP) and 9 in blast crisis (BC). Among the CP pts 4/20 were enrolled because of toxicity to prior imatinib treatment. WT1 and Bcr-Abl kinetics were expressed in relation to Abl expression. Data points were taken at baseline and every 4 weeks thereafter. Quantitative PCR was carried out by Taqman-analysis.
Results: The median duration of nilotinib treatment was 8 months in CP pts, 4 in AP pts and 5.5 in BC pts. The follow up of all pts ranged between 3 and 13 months. All pts received imatinib prior to nilotinib and were Bcr-Abl positive at study entry. In CP samples WT1/Abl ratios (median:1.24x10−3) were 38 fold lower than Bcr-Abl/Abl (median:4.67x10−2) ratios at baseline (p≤0.014, t-test). However, in AP and BC pts no relevant difference of WT1/Abl and Bcr-Abl/Abl baseline levels were detectable. In BC patients a more rapid decline of WT1/Abl ratios compared to Bcr-Abl/Abl ratios was observed within the first four months of nilotinib therapy. In addition, in BC pts elevation of WT1 levels preceded hematologic relapses and was detectable earlier than elevation of Bcr-Abl/Abl ratios.
Conclusions:In contrast to AP and BC pts in CP pts WT1/Abl ratios were significantly lower than Bcr-Abl/Abl ratios confirming the restriction of WT1 expression to early progenitors.In BC pts under nilotinib therapy WT1/Abl ratios may be an early marker of response as compared to Bcr-Abl/Abl ratios.A stable correlation of WT1/Abl and Bcr-Abl/Abl ratios was observed in AP pts under treatment.
FLT3 Mutations in a Large Series of Patients with Acute Promyelocytic Leukemia Treated with All-Trans Retinoic Acid and Anthracycline Monochemotherapy.
Authors: Montesinos Pau , Gonzalez Marcos , Calasanz Maria , Bolufer Pascual , Colomer Dolors , Roman Jose , Gomez Maria Teresa , Martinez Joaquin , Amutio Elena , Sanz Miguel 
Published: October 2019

Abstract
Background: The prognostic significance of FLT3 internal tandem duplication (ITD) and D835 mutations in acute promyelocytic leukemia (APL) remains unclear. Recent studies have revealed an association between FLT3 mutations and leukocytes > 10x109/L, M3-variant subtype and BCR3 isoform. It has been also suggested that patients with FLT3 mutations have a higher induction death rate and a shorter survival. Objectives: To analyze in patients with newly diagnosed APL treated with an ATRA plus anthracycline-based protocol if the presence of FLT3 ITD and D835 mutations influences the clinical and biological presenting features, and the disease outcome.
Material and methods: Between 1997 and 2005, 733 consecutive patients were enrolled in the PETHEMA LPA96 and LPA99 trials. Treatment consisted of Induction therapy with oral ATRA (45 mg/m2/d) and intravenous idarubicin (12 mg/m2/d x 4 days) followed by three courses of consolidation with anthracycline monochemotherapy. In the LPA99 trial, ATRA was added in each cycle of consolidation for intermediate and high risk patients (according to Sanz score). In both trials, maintenance therapy consisted of intermittent ATRA and low dose chemotherapy with methotrexate and 6-mercaptopurine. Using RT-PCR, analysis of the FLT3 ITD and D835 mutations was available in 251 and 191 patients respectively.
Results: Overall, 61 patients (24%) had a FLT3 ITD mutation and 20 patients (10%) had a FLT3 D835 mutation. Only one patient had both mutations. FLT3 ITD was associated with BCR3 isoform (78% vs 32%, p < 0.01), leukocytes > 10x109/L (64% vs 17%, p < 0.01), M3v subtype (46% vs 13%, p < 0.01), bone marrow blasts > 70% (95% vs 80%, p < 0.01) and expression of CD2 and CD34 surface antigens (41% vs 14% and 38% vs 9%, respectively, p < 0.01). FLT3 D835 was not associated with leukocytes > 10x109/L (35% vs 25%, p=0.31) or any other biological characteristic. There was a trend towards a higher induction death rate in patients with FLT3 ITD or D835 mutations, when compared to patients with wild-type FLT3 (15% vs 16% vs 6%, respectively, p=0.08). There was no significant difference on relapse-free survival (RFS) and overall survival (OS) at 7 years in patients with FLT3 ITD, when compared to those with FLT3 D835 or wild-type FLT3 (81% vs 100% vs 87% for RFS, p=0.17; and 76% vs 84% vs 81% for OS, p=0.37).
Conclusion: In a large cohort of patients with newly diagnosed APL, the incidence of FLT3 ITD and FLT3 D835 mutations was 24% and 10%. This study was only able to demonstrate an association of FLT3 ITD with leukocytes > 10x109/L, M3v subtype, BCR3 isoform, and expression of CD2 and CD34 surface antigens, but this mutation had no independent prognostic value in patients with APL receiving state-of-the-art treatment.
Acute Promyelocytic Leukemia Stem Cell Derived from Leukemic Progenitor.
Authors: Rosen Galit P. , Kogan Scott C. 
Published: October 2019

Efforts to identify the leukemic stem cell (or leukemia-repopulating cell, LRC) of acute myeloid leukemia (AML) have led to considerable insights, but also to a number of unresolved questions. While the LRC is believed to arise primarily from the hematopoietic stem cell, data from chronic myelogenous leukemia blast crisis and acute promyelocytic leukemia (APL) suggest that they may arise from more committed cells.
In order to clarify the potential of committed progenitors to become LRCs in some human AMLs, we utilized the MRP8-PML/RARA transgenic murine model of APL to perform initial studies directed at the identification of the LRC in APL. Unsorted spleen cells of MRP8-PML/RARA leukemic mice injected into cohorts of sublethally irradiated mice using limiting dilutions showed that the LRC of APL is present in 1:100 to 1:300 cells. FACS analysis of six primary and serially transplanted leukemic specimens revealed that normal hematopoetic stem cells and myeloid progenitors were rare, comprising .007% and .078% of total splenic cells, respectively. These data raised the possibility that the LRC is present at higher frequency than these normal progenitors and does not belong to any of these populations.
Given the scarcity of normal progenitors, we surmised that the LRC may be part of the leukemic population. We performed FACS analysis of seven primary leukemias to elucidate potential heterogeneity within the leukemic cell population. Myeloblasts had the immunophenotype lymph−Gr1+ckit+CD16/32+CD34mod/+. A smaller population that made up 1.4–12.7% of all cells was lymph−Gr1−ckitlo/mod and had variable expression of CD16/32 and CD34. Such cells have been identified as myelomonocytic cells and represent a stage in myelopoiesis beyond the GMP. Our findings suggest a hierarchy in which Gr1−ckitlo/mod leukemia repopulating cells proliferate and differentiate to maintain the population of abnormal promyelocytes.
We infer from the dilution analysis data that the LRC in this MRP8-PML/RARA mouse model of acute promyelocytic leukemia cannot be a hematopoietic stem cell or a normal myeloid progenitor cell. Also, it is apparent that the entire blast population is not capable of transferring disease; otherwise, all the mice injected would have developed leukemia. While our data does indicate that while leukemogenesis in APL is a hierarchy, it originates from a newly identified leukemia progenitor cell that is Gr1−ckitlo/mod that may be more mature than a GMP. It is quite likely that the LRC lies within this population of cells. Future work will further delineate the immunophenotype of the LRC in this model and in human leukemia, as well as molecular abnormalities that are important in differentiating from the LRC into the myeloblast phenotype.
Comparison of Three Different NOD/SCID-Related Strains in Preclinical Models of Acute Leukemia.
Authors: Agliano Alice , Martin-Padura Ines , Merighetti Paola , Mancuso Patrizia , Rabascio Cristina , Shultz Leonard D. , Bertolini Francesco 
Published: October 2019

Abstract
Transplantation of human acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) primary cells and cell lines in a variety of different strains of immunodeficient mice has led to preclinical models extensively used to investigate AML and ALL stem cells, biology, and drug sensitivity. We investigated the engraftment kinetics of two AML cell lines (HL-60 and KG-1), two ALL cell lines (MOLT-16 and 697) and AML primary cells from an AML M4 patient (QD1-EIO, described in Fusetti et al, Cancer Res 2000) in 3 different strains of NOD/LtSz-Prkdcscid (NOD/SCID, NS)-related immunodeficient mice. NS, NS/beta2 null (NSB) and NOD/SCID/IL-2Rgamma null (NSG) mice were injected ip with 10x106 AML or ALL cells. Mice were observed daily and sacrificed when leukemia-related symptoms were evident. Overall, leukemia-related symptoms were observed in 71, 84 and 86% of NS, NSB and NSG mice, respectively (n=42, p<0.01), after a median of 53, 49 and 35 days (p<0.001). Leukemia engraftment was investigated in the marrow, the blood and the spleen by means of morphology, flow cytometry and quantitative PCR for human genes. AML HL-60 and KG-1 cells accounted for 10-6, 5-1 and 7-3 % of peripheral blood cells in NS, NSB and NSG mice, respectively. ALL MOLT-16 and 697 cells accounted for 2-12, 22-27 and 1-27 % of blood cells in NS, NSB and NSG mice, respectively. AML primary cells QD1-EIO accounted for <1, <1 and 3% of blood cells in NS, NSB and NSG mice, respectively. Similar engraftment results were observed in the marrow and in the spleen. Leukemia cell-injected NSG mice, compared to NS and NSB, showed a significantly higher increase of circulating endothelial mature cells (CEC, enumerated by flow cytometry as CD45−, CD13+ VEGFR2+ cells) and progenitors (CEP, enumerated by flow cytometry as CD45−, CD13+ VEGFR2+ CD117+ cells, see Shaked et al, Cancer Cell 2005). This CEC and CEP increase paralleled leukemia engraftment. Taken together, our data indicate that the 3 different strains have significantly different leukemia engraftment behavior and kinetics. Engraftment in NSG mice is significantly faster compared to the other two strains, leukemia-related microenvironment is differently modulated, and less leukemic burden might be needed to observe leukemia-related symptoms. These data might have major implications to design future studies on leukemia-initiating stem cells, leukemia biology and preclinical leukemia treatment studies.
TRAF6 and C-myb Show Novel Functions in the Nucleus of Lymphoma B Cells.
Authors: Pham Lan V. , Lin-Lee Yen-Chiu , Zhou Hai-Jun , Tamayo Archito T. , Yoshimura Linda C. , Ford Richard J. 
Published: October 2019

Abstract
The tumor necrosis factor (TNF) family (TNF-R; CD40; BAFF-R) plays a key role in neoplastic as well as normal B cell growth and survival mechanisms. TNF receptor-associated factor-6 (TRAF-6) is an adapter molecule that regulates several important signaling pathways critical for cell growth and cell survival. It is a member of seven closely related TRAF proteins that serve as signaling molecules, coupling to TNF-receptor superfamily to intracellular signaling, particularly in the CD40 Signalosome. TRAF6 has shown to be over-expressed and play an important role in cell growth and cell survival through the activation of the key transcription factor NF-kB in aggressive non-Hodgkin’s lymphoma B cells (NHL-B), common B cell neoplasm that have been increasing in recent years. Although much of TRAF-6 functions have focused primarily as an adaptor molecule in signaling pathways in the cytoplasm, the role of TRAF-6 in other cellular compartments has not been investigated. Here, we demonstrate, by confocal microscopy as well as cellular fractionation studies that TRAF-6 resides not only in the cytoplasm but also in the nucleus of lymphoma B cells. Immunoprecipitation studies show that TRAF6 is auto-ubiquitinated in the cytoplasm but not in the nucleus, suggesting that nuclear TRAF6 functions differently than cytoplasmic TRAF6. Chromatin immunoprecipitation (ChiP) cloning assays using anti-TRAF6 polyclonal antibody reveal over 200 clones, one of which contains a 130 bp fragment belonging to the proximal 5′ end of the c-myb oncogene promoter. Further experiments demonstrate that nuclear TRAF6 co-localized with SUMO1 and c-myb, suggesting that TRAF-6 may enter the nucleus through SUMO1 interaction and serve as an E3 sumo ligase, in addition to its known adapter role in cytoplasmic signaling. Over-expression studies show that TRAF6 enhances c-myb sumoylation in lymphoma B cells, where this oncogene is over-expressed. C-myb correlates with TRAF6 protein and mRNA expressions in NHL-B cells, suggesting that TRAF6 may be involved in the modulation of c-myb expression through sumoylation, regulating key genes that are regulated by c-myb. Small interfering RNA (siRNA) targeting c-myb results in inhibition of lymphoma cell survival, suggesting that SUMO1/TRAF6/c-myb interactions are important in cell survival pathways in aggressive NHL-B. Such pathways could represent novel targets for the development of therapeutic agents for aggressive B cell lymphomas.
Regulation of pri-miRNA BIC Transcription and Processing in Burkitt Lymphoma.
Authors: van den Berg Anke , Kluiver Joost , de Jong Debora , Blokzijl Tjasso , Harms Geert , Jacobs Susan , Poppema Sibrand , Kroesen Bart-Jan 
Published: October 2019

Abstract
BIC a primary microRNA (pri-miR-155) transcript is highly expressed in the vast majority of Hodgkin lymphoma and diffuse large B cell lymphoma cases. High miR-155 levels are also observed in these B cell malignancies. In contrast, only very low or no expression of BIC and miR-155 are observed in Burkitt lymphoma cases. In the normal lymphoid system, BIC expression is observed in a proportion of the germinal center B cells and we have previously shown induction of BIC expression by BcR stimulation.
In this study we show the crucial involvement of PKC and NF-κB in the regulation of BIC expression upon BcR triggering of the Burkitt lymphoma cell line, Ramos, which is commonly used as a model system for germinal centre B cell stimulation. Surprisingly, Northern blot analysis did not reveal any miR-155 expression upon induction of BIC expression whereas other microRNAs were clearly detectable. Ectopic expression of BIC in Ramos and HEK293 cells resulted in miR-155 expression in HEK293 but not in Ramos cells suggesting a specific block of BIC to miR-155 processing in Ramos. In line with the results obtained with Ramos, lack of miR-155 expression after induction of BIC expression was also observed in other Burkitt lymphoma cell lines, indicating a generic and specific blockade in the processing of BIC in Burkitt lymphoma. In contrast, induction of BIC expression in normal tonsillar B cells resulted in very high levels of miR-155 expression and induction of BIC expression in Hodgkin lymphoma cell lines also resulted in elevated levels of miR-155. More recent reports have indicated that miRNA processing may be regulated by editing of pri-miRNA sequences by adenosine deaminases acting on RNA. ADARs can convert adenosines to inosines in stemloop sequences which may interfere with proper recognition and cleavage of pri-miRNA by the microprocessor complex. To evaluate whether ADAR editing might explain the observed differences in BIC to miR-155 processing, we analyzed the miR-155 sequence including 100nt up- and downstream of BIC RT-PCR products from stably transfected Ramos cells and PMA/Ionomycin stimulated CA-46 Burkitt lymphoma cells and compared those with sequences obtained from stably transfected HEK293 cells and L1236 cells. Analysis of 30 individual clones per cell line revealed no or very few conversions in the BIC transcripts of the four cell lines. This indicates that ADAR editing is not responsible for the observed BIC processing blockade in BL.
Our data provide evidence for two levels of regulation for mature miR-155 expression: one at the transcriptional level involving PKC and NF-κB, and one at the processing level. Burkitt lymphoma cells not only express low levels of BIC but also prevent processing of BIC via an, as yet, unknown mechanism.
Impact of IL-10 Polymorphisms in Survival of Lymphoid Neoplasms.
Authors: Domingo-Domenech Eva , Benavente Yolanda , Gonzalez-Barca Eva , Montalban Carlos , Guma Josep , Wang Sophia S. , Whitby Denise , de Sevilla Alberto Fernandez , Rothman Nathaniel , de Sanjose Silvia 
Published: October 2019

Abstract
BACKGROUND
Polymorphisms in interleukin-10 (IL-10) genes can influence immune responses that may affect the outcome of lymphoid neoplasms.
METHODS
We analyzed two single-nucleotide polymorphisms (−1082 and −3575) in the IL-10 promoter region in 472 consecutive non-immunessuppresed cases diagnosed with lymphoid neoplasms. Genotypes were tested for an association with overall survival and classical prognostic factors by multivariate analysis, adjusted for clinical characteristics and other confounders. Haplotypes were reconstructed using haplo.stats package and their implications analyzed in lymphoma survival by multivariate analysis.
RESULTS
In our series, patients with IL-10-3575AA genotype had a better overall survival (p= 0.002) for all lymphoid neoplasms as well as within the non-Hodgkin lymphoma patients (p=0.04). We also found that patients carrying the IL-10-1082GG genotype presented a better median overall survival when compared to genotypes IL-10-1082AA/AG. (p=0.05). When information on both genotypes were included in a multivariate analysis, IL-10-3575AA was the only genotype identified as an independent prognostic factor for survival (HR=0.2, 95%CI 0.05–0.92).
Haplotype analysis showed that patients with A-G/A-G diplotype had a longer overall survival (p=0.0042) and it appeared to be an independent prognostic factor for survival (HR:0.26; 95%CI 0.08–0.83).
CONCLUSIONS
IL-10-3575AA and IL-10-1082GG genotypes, together with A-G/A-G diplotype were identified as markers of a favorable lymphoma survival.
Aberrant Methylation of SHP-1 (SH2-Containing Phosphatases 1) and SOCS-1 (Suppressor of Cytokine Signalling 1) Genes in Immunodeficiency-Related Lymphomas.
Authors: Rossi Davide , Deambrogi Clara , Lunghi Monia , Cerri Michaela , Garbellotto Sara , Spina Valeria , De Paoli Lorenzo , Riccomagno Paola , Gloghini Annunziata , Lucioni Marco , Capello Daniela , Paulli Marco , Carbone Antonino , Gaidano Gianluca 
Published: October 2019

Abstract
SHP-1 and SOCS-1 are members of a family of negative regulators of signaling that act downstream of cytokine receptors, receptor tyrosine kinases and receptor complexes of the immune and hemopoietic system. SHP-1 and SOCS-1 act through the inhibition of the JAK2/STAT pathway and their inactivation is considered to be a relevant mechanism in JAK2/STAT pathway induction. Several observations point to induction of the JAK/STAT pathway as an important mechanism of lymphomagenesis. Since DNA methylation is a mechanism of gene silencing involved in the pathogenesis of human cancers, including lymphoma, here we tested the involvement of SHP-1 and SOCS-1 methylation in immunodeficiency-related lymphoma. Tumor samples from 34 HIV-related non-Hodgkin lymphoma (HIV-NHL) and 25 post-transplant lymphoproliferative disorders (PTLD) were analyzed for SOCS-1 and SHP-1 methylation by methylation-specific polymerase chain reaction. The tumor panel included 15 HIV-diffuse large B cell lymphoma (HIV-DLBCL), 10 HIV-Burkitt lymphoma (HIV-BL), 9 HIV-primary effusion lymphoma (HIV-PEL), 10 PTLD-immunoblastic (PTLD-IB), 7 PTLD-centroblastic (PTLD-CB), 4 polymorphic PTLD (P-PTLD), 3 PTLD-BL and 1 PTLD multiple myeloma (MM). Among HIV-NHL, SHP-1 methylation occurred in 23/34 (67%) cases, including 12/15 (80%) HIV-DLBCL, 4/10 (40%) HIV-BL, and 7/9 (78%) HIV-PEL. SOCS-1 methylation occurred in 6/34 (17%) HIV-NHL, including 5/9 (55%) HIV-PEL and 1/15 (7%) HIV-DLBCL. When considering EBV status, SHP-1 was methylated in 10/11 (91%) EBV-negative HIV-NHL and in 12/22 (54%) EBV-positive HIV-NHL. Among PTLD, SHP-1 methylation was detected in 20/25 (80%) cases, including 7/10 (70%) PTLD-IB, 6/7 (86%) PTLD-CB, 3/4 (75%) P-PTLD, 2/3 (66%) PTLD-BL and 1 PTLD MM. SOCS-1 methylation was detected in 3/25 (12%) PTLD, including 1/10 (10%) PTLD-IB and 2/7 (28%) PTLD-CB. When considering EBV status, SHP-1 was methylated in 12/13 (92%) EBV-negative PTLD and in 6/11 (54%) EBV-positive PTLD. The implications of our data are threefold. First, SHP-1 methylation is involved in the majority of HIV-NHL and PTLD. Second, it is remarkable that virtually all EBV-negative HIV-NHL and PTLD carry SHP-1 methylation. In EBV-positive B-cells, EBV infection activates the STAT pathway. It is conceivable that, in EBV-negative HIV-NHL and in EBV-negative PTLD, SHP-1 inactivation through aberrant methylation may surrogate EBV infection for STAT activation. Third, similar to observations in NHL of the immunocompetent host, SOCS-1 methylation is rarely implicated in the pathogenesis of immunodeficiency-related NHL. This notion is supported by the phenotype of SOCS1-deficient mice, that die of a myeloproliferative disease but do not develop a lymphoproliferative disease.