bluuebunny/crossref_metadata_2025_split huggingface dataset:
PLAGL2 Cooperates in Leukemia Development by Direcly Inducing c-Mpl Expression.
Authors: Landrette Sean F. , Castilla Lucio H. 
Published: April 2020

Abstract
Inversion of chromosome 16 is found in 12% of human acute myeloid leukemia (AML). This inversion creates the CBFΒ-SMMHC fusion protein, which hinders myeloid differentiation and triggers AML in collaboration with additional mutations. We have previously shown that the zinc finger transcription factor PLAGL2 efficiently cooperates with CbfΒ-SMMHC in murine AML, and that PLAGL2 transcript levels are significantly increased in inversion 16 human AML. The mechanism of PLAGL2 action in AML, however, is not understood.
In order to identify candidate PLAGL2 target genes in AML, we performed gene expression profiling of primary hematopoietic progenitors expressing CbfΒ-SMMHC and PLAGL2, as well as in AML samples induced by CbfΒ-SMMHC and PLAGL2. The expression of c-Mpl transcript was increased 8 to 32 fold in progenitors and AML samples when compared to controls not expressing PLAGL2. This increase was confirmed by quantitative PCR analysis and correlated with increased c-Mpl receptor in membrane by flow cytometry. Analysis of the c-Mpl proximal promoter identified three conserved Plag binding sites. PLAGL2 binding to these sites was confirmed by chromatin immunoprecipitation and luciferase reporter assays. Furthermore, PLAGL2/CbfΒ-SMMHC AML cells show a significant increase in sensitivity to the c-Mpl-ligand thrombopoietin when compared to non-PLAGL2 CbfΒ-SMMHC-AML samples as measured by Jak2 phosphorylation.
To test whether PLAGL2 induces AML by increasing c-Mpl levels, AML development was analyzed in bone-marrow transplantation assays using CbfΒ-SMMHC-expressing donor cells transduced with c-Mpl expressing retrovirus. All transplanted mice (8/8) succumbed to leukemia with a median latency of 8 weeks, while control groups remained unaffected for 16 weeks. The pathology of Mpl/CbfΒ-SMMHC AML was immuno-phenotypically and morphologically identical to that of PLAGL2/CbfΒ-SMMHC AML.
This study demonstrates that PLAGL2 directly induces c-Mpl receptor expression, and that such increase cooperates with CbfΒ-SMMHC in AML development. These results suggest that AML can result from the alteration of upstream regulators of Mpl/Jak2 signaling.
Trends and Socio-Economic Inequalities in Cancer Survival of Patients with Hodgkin’s Lymphoma Diagnosed in England and Wales between 1986 and 1999.
Authors: Bohlius Julia , Rachet Bernard , Nolte Ellen , Engert Andreas , Coleman Michel 
Published: April 2020

Abstract
Background: Similar to other cancers in England and Wales, survival from Hodgkin’s lymphoma is influenced socially patterned, with those in the most deprived groups showing worse survival rates. Survival from Hodgkin’s lymphoma increased during the 1980s, however, so did the gap in survival between the most affluent and most deprived groups. It is not known whether the suggested widening of this gap continued into the 1990s.
Objective: To examine trends in relative survival (RS) from Hodgkin’s lymphoma in England and Wales diagnosed between 1986 and 1999. It explores how survival varies by socio-economic background and how these differences change over time.
Methods: Anonymized data of 14,831 patients with Hodgkin’s lymphoma diagnosed in England and Wales between 1986 and 1999 and followed up until the end of 2001 were analyzed. We estimated 5-year relative survival (ratio of observed survival of cancer patients and survival that would have been expected if the patients had had the same age- and sex-specific mortality in each time period as the general population). We examined the periods 1986-1990, 1991–1995 and 1996–2001. Patients were assigned to one of five deprivation categories based on the area of residence, from ‘affluent’ to ‘deprived’ using both the Carstairs score (Carstairs 1995) and the ward income domain score, a subcomponent of the indices of multiple deprivation. We estimated differences in survival between the deprivation categories using linear regression, weighted by the variance of the relative survival estimate. Men and women were analyzed separately.
Results: Among men 5-year relative survival improved consistently and statistically significant from 76% in the late 1980s to 83% in the early 1990s. The 5-year relative survival improved on average by 4.23% (95% CI 1.22 to 7.25, p=0.007) every five years. For women there was no evidence that RS improved over time (RS 0.89, 95% CI −3.04; 4.82, p-value = 0.66). For both men and women there was evidence for a deprivation gap in survival. Relative survival at 5 years was on average 5% percent less in men from poor (‘deprived’) areas compared to men from wealthy (‘affluent’) areas (RS −4.9; 95% CI −9.5; −0.39, p=0.036). In women the RS at 5 years was 6% less in women from poor areas compared to women from wealthy areas (RS −6.12; 95% CI −12. to −0.24, p=0.04). There is no conclusive evidence whether the survival gap by deprivation changed over time.
Conclusion: Relative survival of patients diagnosed with Hodgkin’s lymphoma in England and Wales improved only for men, but not for women. The existing deprivation gap in relative survival for both men and women confirms that cancer survival depends on socio-economic background and is inequitable. More research is required to better understand how socio-economic factors affect survival from Hodgkin’s lymphoma.
Emp Null Mice Are Non-Viable and Exhibit Erythroid Differentiation Defect.
Authors: Soni Shivani , Bala Shashi , Gwynn Babette , Sahr Kenneth E. , Peters Luanne L. , Hanspal Manjit 
Published: April 2020

Abstract
Emp, erythroblast macrophage protein, was originally detected in erythroblasts and macrophages, which form erythroblastic islands during erythropoiesis in the human bone marrow. The physical contact between erythroblasts and macrophages was suggested to promote the terminal maturation of erythroblasts, leading to their enucleation in vitro. To evaluate the function of Emp in vivo, we employed gene targeting studies to develop an Emp(−/−) mouse model. Mouse embryonic stem cells containing a gene-trap insertion in Emp were obtained from BayGenomics. Insertion of the gene-trap vector into Emp was verified by direct sequencing of cDNA obtained by 5′RACE. Chimeric mice generated by blastocyst microinjection were intercrossed, and the offspring were genotyped by PCR and Southern hybridization. The Emp (+/−) mice were healthy and fertile. However, no live Emp (−/−) mice were found among the progeny of the Emp (+/−) intercrosses. Analysis of timed pregnancies revealed that Emp (−/−) embryos were present at a frequency roughly consistent with Mendelian inheritance throughout the embryonal stages. Homozygous Emp (−/−) embryos were small and pale compared to their littermates, and they survived embryonic development but died at birth. To determine the effect, if any, of Emp gene deletion on definitive hematopoiesis, livers of +/+, +/−, and −/− embryos at E15.5 were examined after H&E and Giemsa staining of paraffin-embedded serial sections, and cytospins. We found few mature erythroid cells in the sinusoids of homozygotes, in contrast to those of either wild-type or heterozygotes, where abundant enucleated red blood cells were observed. Although nucleated erythrocytes were found in both wild-type and mutant embryos, their relative proportions were very different: the less mature forms (proerythroblasts) predominated in the −/− embryos whereas the more mature forms (polychromatophilic/orthochromatic and enucleated erythrocytes) were most common in +/+ and +/− embryos. Furthermore, erythroblastic islands consisting of a central macrophage surrounded by developing erythroblasts were seen in the cytospin preparations of wild-type and heterozygote livers but not in those of homozygous null livers. Since fetal liver macrophages (FLMs) are indispensable for definitive erythropoiesis, we investigated the effect that Emp’s absence might have on development of FLMs. The E15.5 fetal liver sections were stained with the macrophage-specific F4/80 antigen. Numerous F4/80-positive macrophages were present throughout the liver of normal embryos whereas, the number was substantially reduced in Emp (−/−) liver. In summary, in the absence of Emp, FLMs are significantly reduced and terminal maturation of erythroid cells is negatively affected. Thus, the availability of Emp(−/−) embryos will provide a unique experimental model to study the function of macrophages in definitive erythropoiesis.
Recent Interim Analysis of the HD11 Trial of the GHSG: Intensification of Chemotherapy and Reduction of Radiation Dose in Early Unfavorable Stage Hodgkin’s Lymphoma.
Authors: Diehl Volker , Brillant Corinne , Engert Andreas , Mueller Ralf-Peter , Eich Hans-Theodor , Mueller-Hermelink Konrad , Herrmann Richard , Markova Jana , Ho Anthony , Hiddemann Wolfgang , Doerken Bernd , Greil Richard , Josting Andreas , Pfistner Beate 
Published: April 2020

Abstract
Background: Combined modality treatment consisting of chemotherapy (CT) followed by involved field radiotherapy (IF-RT) is the standard treatment for early unfavourable Hodgkin’s lymphoma (HL). Despite high complete remission (CR) rates, failures are common. We thus compared the baseline-dose BEACOPP regimen with ABVD and 20 with 30 Gy IF-RT in a prospectively randomized trial (HD11) in an attempt to improve outcome in this group of patients.
Methods: Between May 1998 and January 2003, 1570 patients (pts) aged 16–75 with untreated intermediate stage HL (CS I, IIA with risk factors or IIB with elevated ESR and/or ≥3 nodal areas only) were randomized according to a factorial design between 4 cycles of ABVD followed by 30 Gy IF-RT (arm A - standard treatment), 4 ABVD + 20 Gy IF-RT (arm B), 4 baseline-dose BEACOPP + 30 Gy IF-RT (arm C) and 4 baseline-dose BEACOPP + 20 Gy IF-RT (arm D).
Results: In the fifth preplaned interim analysis, 1293 pts were evaluable for the chemotherapy comparison and 1274 for the radiotherapy comparison. Patient characteristics were well balanced between the treatment arms. 95% of patients treated reached CR, 2% had pogressive disease, 8% relapsed and the total mortality rate was 4% with no significant differences between treatment arms for either endpoint. The most frequent haematological toxicities during chemotherapy were leucopenia observed in 32% of pts (ABVD: 25%, BEACOPP: 39%) and anemia in 4% of pts (ABVD &lt;1%, BEACOPP 7%). Infection rate was 5% (ABVD 3%, BEACOPP 7%). The most frequent toxicity during radiotherapy was dysphagia in 5%. 14 secondary neoplasias were observed: 2 AML, 4 NHL, 8 solid tumors with no significant differences between treatment arms.
After a median observation time of three years, freedom from treatment failure (FFTF) was 87% (95%-CI 85–89) and overall survival (OS) was 96% (95%-CI 95–97). Both for FFTF and OS, there was no sequential significant difference either between ABVD (FFTF 87%, OS 97%) and BEACOPP (FFTF 88%, OS 96%) nor 30 Gy (FFTF 90%, OS 97%) and 20 Gy IF-RT (FFTF 87%, OS 97%).
Conclusions: At three years of median observation time, no sequential significant differences in treatment outcome were detected, neither between chemotherapy regimens nor between the different doses of radiotherapy, despite more relapses in 20 Gy radiotherapy arms.
Nuclear Dynamics and Gene Activation during Erythroid Maturation.
Authors: Bender M. A. , Ragoczy Tobias , Byron Rachel , Telling Agnes , Groudine Mark 
Published: April 2020

Abstract
We have investigated the relationships among nuclear positioning, association with RNA polymerase II (PolII) and expression of the murine β-globin locus during erythroid differentiation, as well as the role of the locus control region (LCR) in these processes. Fetal liver cells from wildtype and LCR-deletion mouse strains were stained with a panel of antibodies, and flow cytometry was used to define and isolate cells from four stages of erythropoiesis spanning pro-erythroblasts (stage 1) to orthochromatic normoblasts and nucleated RBC (stage 4). DNA FISH analyses reveal that with increasing erythroid maturation the β-globin locus is less likely to be located in the nuclear periphery. Immuno-FISH demonstrates that PolII speckles initially are spread diffusely throughout the nucleus, with the exception of extreme periphery. With erythroid maturation the number of speckles decline and become more centrally located. Combined DNA FISH /PolII immuno-staining reveals an increase in co-localization of PolII and β-globin alleles during erythroid maturation. These results are consistent with a model in which the locus is more likely to be in the nuclear periphery and away from PolII foci prior to activation, and with maturation, the locus re-locates more centrally and associates with PolII speckles, leading to β-globin activation. To investigate this model, α and β-globin primary transcript FISH were performed and revealed that while β activation lags behind α, both start at stage 2 and increase with maturation. Notably, activation occurs prior to the increased localization of the locus away from the periphery during maturation, suggesting that localization of the locus away from the periphery may be secondary to the redistribution of PolII during erythroid maturation. Analysis of sorted erythroid cells from mice with a targeted deletion of the LCR reveals less co-localization of the β-globin locus and PolII speckles and loss of re-localization away from the periphery during maturation. Moreover, primary transcript FISH analysis of sorted cells from mice lacking the LCR and those carrying a combined deletion of HS 2 and 3 suggests that the LCR affects the likelihood that an allele is expressed, as well as the amount of transcript generated during periods of expression. Taken together our results suggest that the LCR plays a role in locating to, or stabilization of, interactions between the locus and PolII speckles, increasing the probability of β-globin expression, after which the LCR also affects the rate of transcription.
Identification of Switching-Related Transcription Factors from a Comparison of Cord and Adult Human Blood Reticulocytes Transcriptomes.
Authors: Goh Sung-Ho , Joselyn Matthew , Lee Y. Terry , Reed Christopher , Miller Jeffery L. 
Published: April 2020

Abstract
Hemoglobin switching patterns during human ontogeny are highly correlated with transcription of the alpha and beta globin genes on chromosomes 16 and 11, but the molecular and genetic mechanisms responsible for the switching phenomenon are not yet defined. For a better understanding of post-natal globin gene switching and silencing, the expression profiles from purified reticulocyte mRNA (28 separate clinical samples; 14 cord, 14 adult) were studied using Affymetrix HG-U133 arrays (44,229 probe sets). To validate the transcriptome expression levels derived from the microarrays and to discriminate the differences between cord blood and adult blood reticulocytes, we performed quatitative real-time PCR. Among 21 tested control genes, the ratios of adult-to-cord blood (AB/CB) array hybridization intensities versus PCR copy numbers demonstrated a correlation coefficient of 0.978. Hence, the AB/CB array hybridization intensities provide a reliable description of erythroid transcriptome patterns associated with post-natal hemoglobin switching. Stringent screening criteria were then used to identify genes that may be involved in fetal-to-adult hemoglobin switching. All 44,229 probe sets were filtered to identify genes that demonstrated greater than a five fold difference in AB and CB mean signal intensities as well as consistent differences among the 14 samples from each group. In addition, the screening criteria required high-level signals in either AB or CB (&gt;1000 FU), and t-test p-values between AB and CB &lt;0.0001. 107 switching-related probe sets from 94 genes met these criteria, and over 90% of them were detected at higher levels in CB. When this inventory of probe sets was categorized using Gene Ontology, less than half were assigned a known molecular function. Among those, five transcription factors were identified including LIM domain only 2 (LMO2) and Kruppel like factor 3 (KLF3). Those two transcription factors play known roles in hematopoiesis, and LMO2 is required for embryonic erythropoiesis. Two other genes encode zinc finger proteins, but no description of their expression in hematopoietic tissues has been reported. Finally, a novel gene encoding chromatin remodeling and PHD-finger domains was discovered. Postnatal hemoglobin switching is associated with significant changes in the reticulocyte transcriptome. Among the transcripts demonstrating highly significant shifts in their levels, known and novel transcription factors were identified.
Methylation Status and Polymorphism of Asparagine Synthetase Gene in Acute Lymphoblastic Leukemia Cells.
Authors: Akagi Tadayuki , Yin Dong , Kawamata Norihiko , Bartram Claus R. , Miller Carl W. , Koeffler Phillip H. 
Published: April 2020

Abstract
Asparagine synthetase (ASY) is an enzyme expressed ubiquitously in mammalian cells. Expression of the gene, however, is repressed in acute lymphoblastic leukemia (ALL), and the cells are unable to synthesize sufficient asparagine. Depletion of asparagine by L-asparaginase leads to cell death in ALL; therefore, asparaginase has been used as treatment of ALL. Here, we show the methylation status of ASY gene in ALL clinical samples, and report a novel insertional polymorphism in the gene. The ASY gene has a CpG island located from −313 to +336 including 49 CpG sites. We analyzed the methylation status in the region by sodium bisulfite conversion of genomic DNA followed by PCR analysis. A total of 18 of 22 (82%) bone marrow samples of ALL at diagnosis were methylated in the CpG island. After chemotherapic induced-remission, all the bone marrow samples were unmethylated including the 18 patients whose ALL samples were methylated. Experiments by others using an ALL cell line showed that methylation of the ASY gene silenced the gene. In addition, we discovered either a 14 bp or 28 bp inserted sequence between +83 and +84 located in the 1st intron of the gene. Methylation status of the gene was independent of the insertion. To calculate the frequency of the insertion, 92 ALL samples were analyzed by PCR and sequencing. 69 samples (75%) did not contain the insertion in either allele, 22 samples (24%) contained the insertion in one allele, and only 1 sample (1%) contained the insertion in both alleles. To examine whether the insertion was specific for leukemic cells, we analyzed 71 non-ALL samples. 48 samples (68%) did not contain the insertion in either allele, 12 samples (27%) contained the insertion in one allele, and 4 samples (6%) contained the insertion in both allele. These results showed that the insertion is a germline polymorphism; we are exploring if this insertion changes expression of the gene.
In summary, we found a novel insertional polymorphism in the ASY gene. Also, our data for the first time suggests that ALL cells cannot express ASY because the gene’s CpG island is methylated, thus explaining why ALL cells are sensitive to L-asparaginase. Notably, 18% of ALL samples were not methylated, and these patients might be expected to be resistant to L-asparaginase therapy suggesting that a simple test could be of therapeutic value.
Maternal and Birth Characteristics and Risk of Leukemia in Children with Down Syndrome.
Authors: Roesler Michelle A. , Blair Cindy K. , Spector Logan G. , Robison Leslie L. , Olshan Andrew F. , Ross Julie A. 
Published: April 2020

Abstract
High birth weight (&gt; 4000 g) is a risk factor for childhood leukemia, especially for younger children. Advanced maternal age has also been associated with an increased risk, though less consistently than birth weight. Because children with Down syndrome (DS) are at a 20-fold increased risk of developing acute leukemia than the general population, we evaluated maternal and child birth characteristics that may be associated with leukemia in children with DS. A case-control study of DS and acute leukemia was conducted through the Children’s Oncology Group in North America. From 1997 to 2002, 158 children ages birth-18 years with DS and acute leukemia (including 61 acute myeloid leukemia (AML) and 97 acute lymphoblastic leukemia (ALL)) were enrolled. Children with DS alone (n= 173) were identified through the cases’ pediatric clinics and frequency matched to cases on age. Mothers of cases and controls completed a telephone interview that included questions regarding maternal and birth characteristics. Here, we present data related to singleton pregnancies (165 controls and 151 cases (96 ALL/55AML)). Advanced maternal age (≥ 35 vs. &lt;35) was associated with an elevated risk of leukemia (Odds Ratio (OR)=1.79, 95% Confidence Interval (CI))=1.09–2.94), which was apparent for both ALL and AML. There was also a suggestion of an increased risk for children in the highest tertile (&gt;3260 g) of birth weight (OR=1.56, 95% CI=0.87–2.78) compared to children in the reference group (2850–3260 g). Birth length ≥ 21 inches showed an increased risk for AML cases only (OR=3.06; CI=1.25–7.50). In contrast, increasing birth order was associated with a decreased risk, particularly for AML (OR=0.36; 95% CI 0.14–0.97). This is the first study to evaluate maternal and child birth characteristics in relation to leukemia risk in children with DS. Our results add to the growiA. ng body of evidence that suggests that newborn body size may be related to the underlying etiology of childhood leukemia.