bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Prognostic Impact Of Gene Rearrangements and Protein Expression Of MYC, BCL2 and BCL6 In Young High-Risk Patients With DLBCL
Authors: Horn Heike , Ziepert Marita , Barth Thomas F.E. , Bernd Heinz-Wolfram , Wartenberg Martin , Feller Alfred C. , Klapper Wolfram , Hummel Michael , Stein Harald , Lenze Dido , Hartmann Sylvia , Hansmann Martin-Leo , Möller Peter , Cogliatti Sergio , Pfreundschuh Michael , Trümper Lorenz , Löffler Markus , Ott German , Schmitz Norbert , Rosenwald Andreas 
Published: October 2019

Abstract

Purpose
A number of retrospective analyses have looked into the prognostic implication of MYC, BCL2 and BCL6 rearrangements and protein expression for MYC, BCL2 and BCL6 in patients with diffuse large B-cell lymphoma (DLBCL). Many of these studies suffer from small patient cohorts and differing or unknown treatment strategies (with or without Rituximab) administered to patients under study. Furthermore, the median age of these patients was relatively high. We for the first time report on the prognostic consequences of MYC, BCL2 and BCL6 alterations in younger (18-60 years) high-risk patients (aaIPI 2 or 3) with DLBCL.

Patients and Methods
The MegaCHOEP study (Schmitz et al. Lancet Oncology 2012: 13, 1250) randomized patients to 8xCHOEP-14 or sequential high-dose therapy supported by repeated infusions of autologous stem cells. Both treatment arms included 6 infusions of R (375 mg/ sqm). The median patient age was 48 years, and 27% of patients scored an aaIPI of 3. Paraffin-embedded tumor samples from 112 de novo DLBCL were investigated using immunohistochemistry for MYC, BCL2, and BCL6, and fluorescence in-situ hybridisation (FISH) to detect breaks of MYC, BCL2 and BCL6.

Results
Rearrangements of MYC, BCL2 and BCL6 were detected in 13.6%, 20.7% and 30.9% of DLBCL, respectively. A double or triple hit constellation occurred in 10.8%. Presence of BCL2 breaks (RR=4.7, 95% CI: 1.8-12.2) and MYC breaks (RR=2.4, 95% CI: 0.8-7.5), but not of BCL6 breaks were associated with inferior overall survival in univariate and multivariate analyses adjusted for aaIPI and treatment arm. Protein overexpression of MYC ≥30% (RR=2.4, 95% CI: 0.9-6.5), but not BCL2 (≥60%) or BCL6 (≥30%) indicated inferior overall survival. BCL2 overexpression was associated with inferior EFS (RR=2.2, 95% CI: 0.9-5.5) and PFS (RR=2.8, 95% CI: 1.0-8.2). If the same cutpoint for BCL2 used for a similar analysis in elderly patients treated on the RICOVER-60 trial (Horn et al. Blood 2013) was applied, no differences in EFS or PFS were observed.

Conclusion
Rearrangements of MYC and BCL2 seem to be relevant prognostic factors also in young high-risk patients. The frequencies of MYC and BCL2 rearrangements are not substantially higher than reported for other, mostly elderly patient groups carrying IPI scores from 0 - 5; it seems unlikely, therefore, that the rearrangements described here (completely) explain the poor prognosis of young, high-risk patients. The prognostic role of BCL2, BCL6, and MYC as well as other biological risk factors must be validated in independent data sets.

Disclosures:
No relevant conflicts of interest to declare.
An Essential Role For Transferrin Receptor 2 In Erythropoiesis During Iron Restriction
Authors: Wallace Daniel F , McDonald Cameron J , Secondes Eriza S , Ostini Lesa , Rishi Gautam , Hooper John D , Velasco Gloria , Ramsay Andrew J , López-Otín Carlos , Subramaniam V Nathan 
Published: October 2019

Abstract
Iron deficiency and iron overload are common clinical conditions that impact on the health and wellbeing of up to 30% of the world’s population. Understanding mechanisms regulating iron homeostasis will provide improved strategies for treating these disorders. The liver-expressed proteins matriptase-2 (encoded by TMPRSS6), HFE and transferrin receptor 2 (TFR2) play important and opposing roles in systemic iron homeostasis by regulating expression of the iron regulatory hormone hepcidin. Mutations in TMPRSS6 lead to iron refractory iron deficiency anemia, whereas mutations in HFE and TFR2 lead to the iron overload disorder hereditary hemochromatosis. To elucidate the competing roles of these hepcidin regulators, we created mice lacking matriptase-2, Hfe and Tfr2.
Tmprss6 -/-/Hfe-/-/Tfr2-/- mice had iron deficiency anemia resulting from hepatic hepcidin over-expression and activation of Smad1/5/8, indicating that matriptase-2 predominates over Hfe and Tfr2 in hepcidin regulation. Surprisingly, this anemia was more severe than in the Tmprss6-/- mice, demonstrated by more extensive alopecia, lower hematocrit and significant extramedullary erythropoiesis in the spleen. There was increased expression of erythroid-specific genes in the spleens of Tmprss6-/-/Hfe-/-/Tfr2-/- mice, consistent with the extramedullary erythropoiesis. Expression of Tfr2 but not Hfe in the spleen was increased in the Tmprss6-/- mice compared to wild type and correlated with the expression of erythroid genes, suggesting that Tfr2 is expressed in erythroid cells. Further analysis of gene expression in the bone marrow suggests that the loss of Tfr2 in the erythroid cells of Tmprss6-/-/Hfe-/-/Tfr2-/- mice causes a delay in the differentiation process leading to a more severe phenotype.
In conclusion, our results indicate that Hfe and Tfr2 act upstream of matriptase-2 in hepcidin regulation or in a way that is overridden when matriptase-2 is deleted. These results indicate that inhibition of matriptase-2 would be useful in the treatment of iron overload conditions such as hereditary hemochromatosis. We have also identified a novel role for Tfr2 in erythroid differentiation that is separate from its canonical role as a regulator of iron homeostasis in the liver. This important role of Tfr2 in erythropoiesis only becomes apparent during conditions of iron restriction. Our results provide novel insights into mechanisms regulating and linking iron homeostasis and erythropoiesis.

Disclosures:
No relevant conflicts of interest to declare.
Either Combined-Modality Or Radiotherapy Alone Provide Favorable Outcome In Stage I-II Mantle Cell Lymphoma: A Report Of 82 Patients From The International Lymphoma Radiation Oncology Group (ILROG)
Authors: Dabaja Bouthaina S. , Tsang Richard , Qi Shunan , Allen Pamela , Hodgson David C. , Ricardi Umberto , Hoppe Richard T. , Ng Andrea , Specht Lena , Li Ye-xiong , Terezakis Stephanie , Constine Louis , Yahalom Joachim 
Published: October 2019

Abstract
Patients with Mantle cell lymphoma (MCL) often present with advanced-stage and have a poor prognosis. Although MCL is highly radiosensitive, RT is rarely used other than for palliation. Stage I-II presentation is rare, and there is only limited data on treatment outcome with less aggressive combined modality approaches or RT alone. To assess the outcome of this group of patients, we retrospectively reviewed 82 patients referred to radiation oncology at 9 ILROG institutions between 1990 and 2012. Of the 82 patients, 65 (79%) were males, 44 (54%) were > 60 years; 47 (57%) were stage I, 61 (74%) presented in the head and neck areas, 51 (62%) had extranodal presentation, 14 (17%) were bulky (>5 cm). Treatment approaches consisted of combined modality (CM) with chemotherapy and consolidative radiation therapy (RT) in 66 (80%) and definitive RT alone in 16 (20%). The median radiation dose used was 35 Gy (range 12-45). Chemotherapy consisted of cyclophosphamide, adriamycin, vincristine, and prednisone (CHOP) or CHOP-like, regimens in 62 (57%), rituximab was added to chemotherapy in 51 (62%). The median follow-up was 60 months (range 5-245). At last follow up, 26 (32%) patients relapsed (70% of relapses were outside the radiation field) and 21 (26%) died (2 deaths unrelated to disease or treatment).
The 5-and 10-years overall survivals (OS) were 71% and 63%, respectively. The 5-and 10-years disease free survivals (DFS) were 75% and 39%, respectively.
On univariate analysis, the following characteristics were significantly associated with DFS and OS: age as a continuous variable (HR=1.05, p=0.005 for DFS; HR=1.05, p=0.006 for OS) and bulky disease (HR=2.78, p=0.03 for DFS; HR= 3.18, p=0.02 for OS). On multivariate analysis, both DFS and OS were significantly influenced by: age as a continuous variable (HR=2.09, p=0.03; HR=1.05, p=0.005), bulky disease (HR=2.23, p=0.05; HR=3.1, p=0.03), and female gender only affected DFS (HR=2.16, p=0.03).SurvivalAll (%)Combined modality (%)RT alone up-front(%)5-years OS71689210-years OS6362745-years DFS75738910-years DFS393657

Conclusion
Patients who presented with stage I-II MCL and were treated with programs that included consolidative local radiotherapy or even radiotherapy alone demonstrated favorable disease control and relatively long survival. It appears that radiotherapy as a sole modality can offer in selected patients a comparable DFS and OS to a combined modality approach.

Disclosures:
No relevant conflicts of interest to declare.
Using Forced Chromatin Looping To Overcome Developmental Silencing Of Embryonic and Fetal β-Type Globin Genes In Adult Erythroid Cells
Authors: Rupon Jeremy W , Deng Wulan , Wang Hongxin , Gregory Philip D , Reik Andreas , Dean Ann , Blobel Gerd A. 
Published: October 2019

Abstract
The β-genes undergo developmental activation and silencing in part by competing for their upstream enhancer, the locus control region (LCR). In adult erythroid cells, the LCR contacts the β-globin gene promoter by forming a loop that precludes interaction with the embryonic and fetal β-type globin genes. Reversing this developmental gene expression switch in favor of embryonic/fetal genes has therapeutic implications for patients with hemoglobinopathies. Here we employed a forced chromatin looping approach to activate the silenced murine embryonic βh1-globin gene and the human fetal γ-globin gene in adult erythroid cells. We have previously shown that forced recruitment via artificial zinc finger proteins of Ldb1, a protein necessary for long-range chromatin interactions at the β-globin locus, can trigger chromatin loop formation and transcription initiation. Here, we designed a zinc finger protein targeting the βh1 promoter, fused it to the self-association domain of Ldb1 (βh1-Ldb1), and introduced it into an adult murine erythroid cell line that normally produces nearly 100% adult β-globin. βh1-Ldb1 expression activated βh1-globin transcription up to 3000-fold accounting for ∼20% of total β-globin expression. βh1-Ldb1 similarly increased expression of βh1-globin in fetal liver derived primary erythroid cells. These results are striking given the degree to which murine embryonic globin genes are normally repressed. To test whether the activity βh1-Ldb1 was due to a looped interaction of the βh1 promoter with the LCR, we introduced βh1-Ldb1 into fetal liver derived erythroblasts from mice in which the LCR had been deleted. βh1-Ldb1 was virtually inactive in the absence of the LCR, demonstrating the dependence on the LCR and, by inference, long range looping of βh1-Ldb1 function. We next extended this approach to the human β-globin locus in an effort to activate expression of the fetal γ-globin gene in adult erythroid cells. Ldb1 was fused to a previously described γ-globin promoter binding zinc finger protein, GG1, to generate GG1-Ldb1. Introduction of GG1-Ldb1 into adult primary human erythroid cells strongly activated γ-globin expression with a concomitant reduction in β-globin transcription. Strikingly, γ-globin accounted for nearly 90% of total β-type globin transcription. Furthermore, fetal hemoglobin expression was nearly pan-cellular as determined by flow cytometry. These results demonstrate the power of forced chromatin looping to reprogram developmental regulation of gene expression, and provide a novel proof of concept for activating the γ-globin gene for the benefit of patients with hemoglobinopathies.

Disclosures:
Gregory: Sangamo BioSciences: Employment. Reik:Sangamo BioSciences: Employment.
Primary Cold Agglutinin Associated Lymphoproliferative Disease: A B-Cell Lymphoma Of The Bone Marrow Distinct From Lymphoplasmacytic Lymphoma
Authors: Randen Ulla , Trøen Gunhild , Tierens Anne Maria , Steen Chloe , Abdirashid Warsame , Klaus Beiske , Geir Tjønnfjord , Sigbjørn Berentsen , Delabie Jan 
Published: October 2019

Abstract
Primary chronic cold agglutinin disease (CAD) is a rare hemolytic disease caused by the production of cold agglutinin antibodies that bind to erythrocyte I/i blood group antigens at low temperatures. The binding is mediated by monoclonal IGHV4-34-encoded cold agglutinins. CAD is a disease of older individuals with a median age of 76 years and accounts for about 15 % of all cases of autoimmune hemolytic anemia. The estimated incidence of CAD is 1-4/106 worldwide. The aim of this study is to better characterize the underlying lymphoproliferative disorder in CAD patients by review of bone marrow histology, immunophenotypic and genetic analysis. The bone marrow trephine biopsies of 54 patients with CAD showed circumscribed intraparenchymatous nodules with small monotonous monoclonal B-cells (median infiltration: 10% of marrow cells) with a CD20+, IgMs+, IgDs+, CD27+, CD5-/+, CD11c-/+, CD23-, CD38- immunophenotype. Neither plasmacytoid cytological features nor expression of plasma cell differentiation-associated transcription factors IRF4, BLIMP1 and XBP1 were expressed. However, a limited number of mature monoclonal IgM+, IgD- plasma cells were present outside the lymphoid nodules and were diffusely scattered throughout the marrow. Thus, the bone marrow in CAD shows a characteristic lymphoid infiltration pattern that is different from lymphoplasmacytic lymphoma that typically shows an admixture of lymphoid cells, plasmacytoid cells and plasmacells and that, not infrequently, shows a peritrabecular localisation. Of interest, the MYD88 L265P mutation, typical of lymphoplasmacytic lymphoma, was not detected (17/17 cases, sensitivity 3 %). Somatically mutated monoclonal IGHV4-34 gene rearrangement was demonstrated in eight patients with frozen samples (mean sequence homology 95.4%). Unexpectedly, no mutations of BCL6 intron 1 were demonstrated, except in one patient. BCL6 intron 1 mutations usually accompany immunoglobulin gene mutations in B lymphocytes that have matured in the germinal center. In conclusion, the histological, immunophenotypic and genetic properties of CAD are proof that CAD is a lymphoproliferative disease that is distinct from lymphoplasmacytic lymphoma with which it has previously been confounded. The disease likely originates from B lymphocytes that have not matured in the germinal center.

Disclosures:
No relevant conflicts of interest to declare.
Lenalidomide, Bendamustine, and Rituximab As First-Line Therapy For Patients >65 Years With Mantle Cell Lymphoma: Preliminary Results From The Nordic Lymphoma Group MCL4 (LENA-BERIT) Phase I-II Trial
Authors: Jerkeman Mats , Albertsson-Lindblad Alexandra , Kolstad Arne , Laurell Anna , Räty Riikka , Grønbæk Kirsten , Sundberg Jan , Pedersen Lone Bredo , Geisler Christian 
Published: October 2019

Abstract

Background
Mantle cell lymphoma is a disease of the elderly, with a median age of 70 years. Younger patients may be treated with potentially curative treatment including high dose chemotherapy. For elderly patients, however, no standard therapy has been defined. In the current trial, we investigate if the addition of lenalidomide (LEN) to rituximab (R)+bendamustine (B) (B 90 mg/m2 D1-2 and R 375 mg/m2 D1) followed by maintenance with LEN for 7 months may enhance efficacy, with manageable toxicity, for the older population of MCL patients.

Methods
Eligibility criteria were age > 65 years, or ≤ 65 years, unable to tolerate high dose chemotherapy, with untreated mantle cell lymphoma, stage II-IV. BR was given for 6 cycles q4w. In the phase I part, the MTD of LEN was established as 10 mg days 1-14 during the induction phase, cycles 2-6. Prednisolone 20 mg days 1-14 was given during cycle 2. When LEN was initially given from cycle 1, we encountered unexpected grade III-IV toxicity in the form of cutaneous and allergic reactions. In the maintenance phase, LEN single therapy was given as follows: cycles 7-8 - 10 mg days 1-21, cycles 9-13 - 15 mg days 1-21.

Results
The trial was concluded June 1, 2013, after inclusion of 51 patients, of whom 24 were in the phase I part. The median age is 72 years. According to MIPI, 55% were high risk. Presently, 29 patients are evaluable for response after 6 cycles LBR. ORR is 28/29 (97%), CR+CRu 23 (79%). 17 out of 28 evaluable patients (61%) were MRD-negative after 6 cycles. After a median follow-up of 18 months, the median PFS has not been reached, and the estimated PFS at 2 years is 74%. Eight patients have died, 3 due disease progression, 3 due to treatment related toxicity, 1 of lung cancer in a heavy smoker, 1 of CMML. Overall survival at 2 years is 87%.

Conclusions
When omitted in cycle 1, lenalidomide in combination with R-bendamustine is feasible as first-line therapy in older patients with MCL, and is associated with a high response rate, also as assessed by MRD. The long term efficacy of this regimen remains to be established by longer follow-up.

Disclosures:
No relevant conflicts of interest to declare.
Altered Expression and Mutation Analysis Of Heterogeneous Nuclear Ribonucleoprotein k In Bone Marrow Of Primary Myelofibrosis Patients
Authors: Bozinovic Ksenija , Manshouri Taghi , Post Sean M. , Zhang Ying , Gallardo Miguel , Zhang Xiaorui , Verstovsek Srdan 
Published: October 2019

Abstract
The heterogeneous nuclear ribonucleoproteins (hnRNPs) are RNA/DNA-binding proteins that consist of several family members: A1, A2, B1, C2 and K. hnRNPs have been implicated in numerous cellular processes and when dysregulated, have been suggested to impact tumorigenesis. hnRNP A2/B1 is overexpressed in some lung cancers and has been suggested to be a useful early detection marker for lung carcinoma. hnRNP K has been reported to associate with colon cancer and directly interacts with DNA, RNA, and proteins to regulate gene expression in numerous cellular processes involved in mitogenic responses and tumorigenesis. Loss of hnRNP K expression results in defects in differentiation and apoptosis, and increased hnRNP K expression has been associated with loss of apoptosis and an increase in cellular proliferation.
To explore the possibility of altered hnRNP K expression or mutations in primary myelofibrosis, we isolated mRNA from primary blood or bone marrow mononuclear cells from patients with myelofibrosis (n=62) and healthy controls (n=19). We examined hnRNP K levels and determined that hnRNP K is significantly overexpressed in myelofibrosis (p= <0.0001). Sequencing of the hnRNP K locus revealed a single nucleotide alteration (A-to-G) one allele of intron 5 in 43% of myelofibrosis patients (n= 21) In contrast, only one out of 11 control samples harbored this alteration ((x2 p = 0.05). At present, we are investigating whether G allele is a mutation or a SNP. Furthermore, western blotting indicates an increase in phosphorylation of serine-248 in myelofibrosis patients. Together, these data suggest altered hnRNP K expression and mutations in myelofibrosis that might play a significant role in myeloproliferation and hematopoietic differentiation in myelofibrosis patients. Given the significant impact that hematopoietic differentiation has on leukemogenesis, it is imperative to decipher

Disclosures:
No relevant conflicts of interest to declare.
The PTPN22 R620W Polymorphism Is Not Associated With Clinical and Molecular Correlates Of Chronic Lymphocytic Leukemia (CLL) In Patients Originating From a Region With Low Prevalence Of The Autoimmunity Risk Variant
Authors: Panovska-Stavridis Irina , Trajkova Sanja , Eftimov Aleksandar , Ivanovski Martin , Dukovski Dusko , Marija Popova-Simjanovska , Marija Staninova , Cevreska Lidija , Dimovski Aleksandar 
Published: October 2019

Abstract
PTPN22 is a protein tyrosine phosphatase that modulates antigen-receptor signaling in T and B cells. A PTPN22 R620W polymorphic allele (C1858T, rs2476601) has been associated with increased risk for the development of multiple autoimmune diseases in patients originating from Northwestern Europe, whereas this association is less clear in patients originating from Southern Europe where the 620W (1858T) allele is less frequent. A higher frequency of the 620W risk allele has recently been reported in Chronic lymphocytic leukemia (CLL) patients originating from Northwest Europe. These data adds to the recent findings that PTPN22 is markedly overexpressed in the majority of CLL patients and possibly contributes to the pathogenesis of the disease by affecting apoptosis/survival of antigen stimulated CLL B-lymphocytes.
In order to extend further those observations we conducted a retrospective study on the frequency of the PTPN22 R620W variant in 87 CLL patients and 108 age/sex matched normal controls without a history of malignant disease originating from the Republic of Macedonia (Southeastern Europe) and compared the association of the variant with various clinical/molecular correlates.
Similar allele frequency of the minor T allele (0.06 and 0.063) and genotype distribution (0.88, 0.115 and 0.005 of CC, CT and TT genotypes) was found in CLL patients and normal controls, respectively. Both study groups were in Hardy-Weinberg equilibrium. Also, the PTPN22 R620W variant was not associated with any of the studied clinical and biological parameters, such as blood counts, CD38 expression, IGHV mutation status, incidence of autoimmune hemolytic anemia and autoimmune thrombocytopenia, immunoglobulin levels, time-to-treatment, progression free survival and overall survival.
We conclude that the PTPN22 R620W variant is not a risk factor for the development or progression of CLL in patients originating from the Southeastern European region. The differences of our data from the reported findings for CLL patients originating from Northwest Europe mirrors the differences observed in patients with various autoimmune diseases, suggesting a possible gene-environmental factor that affects the influence of the R620W variant on lymphocyte homeostasis.

Disclosures:
No relevant conflicts of interest to declare.
CD1d Expression Is Higher In Chronic Lymphocytic Leukemia Patients With Unfavorable Prognosis
Authors: Bojarska-Junak Agnieszka , Hus Iwona , Dmoszynska Anna , Rolinski Jacek 
Published: October 2019

Abstract
Chronic lymphocytic leukemia (CLL) is characterized by a very heterogeneous clinical course, which is slow and indolent in most of the patients, however some patient experience rapid disease progression and anticancer therapy is required shortly after the diagnosis. Many issues in CLL development and progression are still unclear. The functional consequences of CD1d expression on tumour cells are not well understood. However, increasing evidence suggests that they may affect iNKT cells.The role of CD1d expression in CLL immunopathogenesis remains undefined. In this study, we investigated the potential role of CD1d in CLL by analyzing the level of CD1d expression on leukemic B cells in peripheral blood of120 patients and assessed its correlation with prognostic markers such as ZAP-70 and CD38 expression, Rai stages and unfavourable cytogenetic changes.Measuring CD1d expression by flow cytometry and qRT-PCR, we showed lower CD1d molecule and CD1d mRNA expression in B cells of CLL patients than of healthy controls. The frequency of CD1d+/CD19+ cells, CD1d staining intensity and CD1d transcript levels increased with the disease stage. CD1d expression was positively associated with ZAP-70 and CD38 expressions as well as with unfavourable cytogenetic changes (17p deletion, 11q deletio),. We established the relationship between high CD1d expression and shorter time to treatment and overall survival. The percentage of CD1d+/CD19+cells inversely correlated with the percentage of iNKT cells. iNKT cells ζ-chain expression was downregulated in the high-CD1d group.These results suggest that high CD1d expression is associated with poor prognosis of CLL and might be involved in disease progression.

Disclosures:
No relevant conflicts of interest to declare.