bluuebunny/crossref_metadata_2025_split huggingface dataset:
New Approaches to the Regulation of Erythropoiesis and Hematoglobin Synthesis in β-Hemoglobinopathies: Studies with the Immunomodulatory Drug CC-4047.
Authors: de Parseval Laure A. Moutouh , Verhelle Dominique , Glezer Emilia , Brady Helen , Corral Laura G. , Morris Christopher L. , Muller George , Chan Kyle 
Published: April 2020

Abstract
Sickle cell disease (SCA) and β-thalassemia constitute worldwide public health problems and new therapies are needed. Inhibition of hemoglobin S (HbS) polymerization is a major target for therapeutic approaches in SCA. New experimental therapies including hydroxyurea (HU) have attempted to augment the synthesis of fetal hemoglobin (HbF) and improve upon current treatment. Clinical trial results have demonstrated that lenalidomide (Revlimid®), recently approved by the FDA, reduces or even eliminates the need for red blood cell transfusions in some anemic myelodysplastic patients. We have examined whether CC-4047, another IMiDs® immunomodulatory drug currently under evaluation for the treatment of hematological cancers could regulate erythropoiesis and hemoglobin synthesis. For this purpose, we used an in vitro culture model to differentiate human erythroid progenitors from bone marrow or peripheral blood CD34+ cells. We demonstrate that CC-4047 is a potent inducer of HbF that synergizes with HU, the sole drug approved for SCA, during erythroid differentiation of CD34 progenitors isolated from healthy donors and patients with SCA. In addition CC-4047 modulates erythropoiesis, slowing erythroid maturation and increased proliferation of immature erythroid cells. Unlike other inducers of HbF such as HU, 5-aza-cytidine and butyrate, CC-4047 is not cytotoxic. The percentage of F-cells are increased by 2 to 3 fold with pan-cellular expression of HbF and an increase of Fetal/Adult hemoglobin ratio that are relevant for SCA. Gene expression profiling of erythroid differentiated cells shows that CC-4047 regulates specific erythroid transcription factors and genes that participate in hemoglobin synthesis, and genes involved in cell cycle and cellular differentiation. CC-4047 controls globin gene expression during erythroid differentiation by increasing the rate of γ-globin and ε-globin transcription and by inducing sustained expression of fetal and embryonic hemoglobin synthesis.
Our results support the hypothesis that CC-4047, alone or in combination with certain current approved therapies comprising HU, can restore effective erythropoiesis and increase the ratio of fetal to adult hemoglobin. Moreover, CC-4047 has demonstrated anti-inflammatory and immunomodulatory effect in vivo that could help to limit the inflammatory state in sickle cell patients. In conclusion, administration of CC-4047 may represent an innovative new therapy for β-hemoglobinopathies.
Phase 2 Study of Bortezomib Weekly or Twice Weekly Plus Rituximab in Patients with Follicular (FL) or Marginal Zone (MZL) Lymphoma: Final Results.
Authors: De Vos Sven , Dakhil Shaker R. , McLaughlin Peter , Saleh Mansoor N. , Belt Robert , Flowers Christopher , Holladay Charles , Knapp Mark , Boral Anthony , Zhang Tracy , Goy Andre 
Published: April 2020

Abstract
Background: Bortezomib (btz, VELCADE®) has single-agent activity in relapsed/refractory non-Hodgkin’s lymphoma (NHL), including FL and MZL, on a twice-weekly schedule. Rituximab (R) is approved for relapsed/refractory CD20+ B-cell NHL. Preclinical studies show additive activity in lymphoma cell lines and lymphoma-bearing SCID mice. Btz and R have no overlapping toxicity. Weekly btz is active and safe in multiple myeloma trials.
Methods: Patients (pts) with measurable CD20+ FL or MZL, KPS ≥50%, and a response with TTP ≥4 months to any prior R-containing regimen, were randomized to btz 1.3mg/m2 twice weekly (d 1, 4, 8, 11 of 21-d cycle, 5 cycles, Arm A) or btz 1.6mg/m2 weekly (d 1, 8, 15, 22 of 35-d cycle, 3 cycles, Arm B). R 375mg/m2 was given weekly for 4 weeks from d 1, cycle 1, in both arms. Response was evaluated by independent radiology review (IRR) and by investigators using the IWRC.
Results: 81 pts (41 Arm A, 40 Arm B) were enrolled; median age: 64 yrs (63.0, 64.5); FL: 86% (80%, 93%); Stage IV: 47% (44%, 50%); KPS &lt;90%: 20% (17%, 23%); LDH &gt;ULN: 27% (32%, 23%); ≥1 extranodal site: 43% (41%, 45%); FLIPI score ≥2: 69% (71%, 68%). In total, 56% (54%, 58%) had ≥2 prior lines of therapy, 81% (80%, 82%) had prior CHOP/CVP ±R, 84% (78%, 90%) had prior R (alone or combination), and 8% (7%, 8%) had prior SCT. Median % of planned cumulative btz dose received: 75% (Arm A) and 99% (Arm B); median total btz: 19.5 and 19.0mg/m2; median dosing intensities per week: 1.6 and 1.3mg/m2. Response rates and TTP in evaluable pts are shown in Table 1. TTP curves by IRR are broadly overlapping between the arms (Figure). Most common AEs were fatigue, GI events, and peripheral neuropathy. Grade ≥3 AEs were more common in Arm A (54%) vs Arm B (35%), notably thrombocytopenia (10% vs 0%) and neutropenia (10% vs 3%), as were grade ≥4 AEs and serious AEs.
Conclusion: Weekly and twice weekly btz + R are active and well tolerated in relapsed/refractory CD20+ indolent NHL. The more convenient weekly regimen appears to offer similar efficacy with less toxicity. Weekly btz 1.6mg/m2 + R is being compared with R in a phase 3 study.
Response rates and time to events data By investigator By IRR Arm A, N=37 Arm B, N=40* Arm A, N=37* Arm B, N=40* *1 pt with no post-baseline assessment Response rate (CR+CRu+PR), n (%) 21 (57) 21 (53) 17 (46) 15 (38) CR+CRu 7 (19) 6 (15) 4 (11) 1 (3) PR 14 (38) 15 (38) 13 (35) 14 (35) SD 9 (24) 13 (33) 8 (22) 17 (43) PD 7 (19) 5 (13) 11 (30) 7 (18) Median TTP, mo 9.9 9.0 5.2 9.7 Figure Figure
Gene Expression Profile of Idiopathic Thrombocytopenic Purpura (ITP) Reveals Elevated Expression of Interferon Regulated Genes.
Authors: Sood Ruchira , Wong Wendy , Gotlib Jason , Jeng Michael , Zehnder James L. 
Published: April 2020

Abstract
Idiopathic thrombocytopenic purpura (ITP) is an acquired disorder of unknown etiology characterized by reduction in platelet counts. To search for mechanisms underlying the pathophysiology of ITP, we used DNA microarrays to determine the ITP whole blood expression profile. We hypothesized that transcriptional changes associated with the low platelet counts will provide us with new markers for improved diagnosis in addition to novel mechanistic information to understand and treat ITP. We analyzed 29 samples from ITP patients with ages ranging from 1–70 years, of which 15 samples were from either acute or chronic ITP patients with platelet counts ranging from 2,000–56,000 X109/L and additional 14 samples were collected from patients at different stages of recovery from ITP. We used Significance Analysis of Microarrays (SAM) to select genes that best distinguished the 15 ITP samples from 5 controls. We identified 490 differentially expressed genes constituting an ITP expression profile (Figure 1). One of the most significant findings was the relatively high expression of several genes known to be induced by Interferon (IFN) in the ITP samples. Elevated expression of interferon induced genes has also been reported in patients with several autoimmune diseases (Baechler, 2006) including SLE (Baechler, 2003; Bennet, 2003 & Crow, 2003) and systemic sclerosis (Tan, 2005) suggesting that this may a common theme underlying autoimmune diseases, including ITP. In addition, we previously found changes in expression of the interferon simulated genes during a cycle of changing platelet counts in two patients with cyclic thrombocytopenia strengthening a connection between interferon signaling and platelet counts. The greatest differential expression at high significance was observed in the interferon induced genes IFIT1 and IRF2 which showed 12 and 8 fold higher expression in ITP samples relative to normal samples. Additional genes found to be expressed at higher levels in ITP included EPSTI1, PARP 14 and INPP1, while genes with reduced expression in ITP were GNG10, PMX1 and G0S2. The ITP gene signature determined by us indicates a significant role for IFN signaling events in both pediatric and adult ITP, and further identifies additional differentially expressed genes. This approach points to a number of new targets to study in the pathogenesis and prognosis of ITP.
Figure Figure
Peripheral T Cell Apoptosis Is Associated with Homeostatic T Cell Proliferation and Plays a Significant Role in Post-Transplant Immune Deficiency.
Authors: Alpdogan Onder , Patel Neel , Suh David , Grubin Jeremy , McGoldrick Suzanne , Smith Odette M. , Lu Sydney , King Christopher , Kochman Adam , Hubbard Vanessa , van den Brink Marcel R.M. 
Published: April 2020

Abstract
Post-transplant peripheral T cell apoptosis is an important (but ill-defined) contributor to the delay in T cell recovery after an allogeneic bone marrow transplantation (BMT). Using murine bone marrow transplantation (BMT) models to analyze post-transplant peripheral T cell apoptosis, we found that donor BM-derived T cells had a higher percentage of apoptotic cells, with decreased BCL-2 levels in young (3-month old) and old (20-month old) recipients of T cell-depleted (TCD) and T cell-repleted allogeneic BMT. Allogeneic recipients of donor BM deficient of Fas, BAX and TRAIL, or with overexpression of BCL-2 and AKT showed no decrease in donor-derived apoptotic T cells, suggesting that these molecules are not directly responsible for post-transplant peripheral T cell apoptosis. However, we noted that both CD4+ and CD8+ Memory T cells (CD44high) exhibited increased apoptosis when compared to naive T cells post-BMT and therefore used RAG-2-eGFP transgenic mice to study whether T cell maturation affects peripheral T cell apoptosis. We found increased numbers of apoptotic cells in mature (eGFPlow) T cells compared to non-divided (eGFPhigh) recent thymic emigrants in RAG-2-eGFP transgenic mice and in recipients of RAG-2-eGFP transgenic (Tg) BM. Interestingly, most of the apoptotic T cells were of eGFPlowCD44high memory phenotype in both CD4+ and CD8+ T cells, while eGFPlowCD44low naive T cells showed no sign of enhanced apoptosis. Moreover, we found similar increased levels of apoptosis of homeostatically proliferating T cells (CD44high-memory-like cells) in recipients of lineage-depleted allogeneic RAG2-eGFP Tg BM or B6 BM. We then tested the effects of homeostatic T cell proliferation on peripheral T cell apoptosis in recipients of syngeneic CFSE-labeled T cells and found again increased levels of apoptosis in the fast proliferating populations of both CD4+ and CD8+ T cells. These cells expressed high levels of CD44 but down-regulated CD62L on their surface and in contrast to allogeneic proliferation exhibited low levels of CD25 expression and high level of IL-7Rα expression (75%). In addition, BCL-2 expression was down-regulated in the fast-proliferating cell population in both CD4+ and CD8+ T cells.
We conclude that newly generated donor BM-derived T cells have increased peripheral T cell apoptosis, which is associated with homeostatic proliferation and does not involve Fas, BAX, TRAIL, and AKT. This mechanism of peripheral T cell apoptosis, which has not been recognized previously, plays a significant role in post-transplant T cell deficiency and represents a promising target for novel strategies to overcome post-transplant immune deficiency in BMT recipients.
Rapid and Massive Expansion of Cord Blood Derived Cytokine Induced Killer (CIK) Cells: An Innovative Proposal for the Treatment of Leukemia Relapse after Cord Blood Transplantation.
Authors: Introna Martino , Franceschetti Marta , Ciocca Alice , Borleri Gianmaria , Conti Elena , Golay Josee , Rambaldi Alessandro 
Published: April 2020

Abstract
Cytokine induced killer cells (CIK) are CD3+/CD56+ T/NK cells with cytotoxic potential against leukemic and other tumor cells but not normal bone marrow in vitro and in vivo. They are expanded in vitro with rhIL-2 after stimulation of peripheral blood mononuclear cells with OKT3 and IFN-γ. We have shown in a recent phase I study that 107/kg allogeneic CIK cells can be safely given to patients relapsing after allogeneic bone marrow transplantation and show evidence of anti-leukemic activity in vivo with very little GVHD. Cord blood (CB) transplantation is progressively becoming an extensively used treatment for patients with malignant disorders. One major limitation of this procedure is the lack of donor derived cells to perform donor lymphocyte infusions in case of relapse. In order to be able therefore to extend the use of CIK cells to the CB transplantation setting, we have standardised a 21 days expansion protocol to produce CIK cells starting from very small amounts of nucleated cells isolated from cord blood. Using this protocol, 15x106 mononuclear cells (MNC) from CB containing a mean 0.3x106 CD3+/CD56+ yielded on average 805 x 106 MNC (50 fold expansion) containing 630 x106 CD3+/CD56+ cells (corresponding to a fold expansion of 1860 for CIK). In order to transfer the method to a clinical setting, we explored the possibility of expanding the residual cells recovered from the empty bags after CB transplantion. Three used CB bags were returned to the laboratory after transplantation and repeatedly washed. An average of 22 x106 nucleated cells could be recovered, yielding a mean 473 x106 CD3/CD56+ cells at the end of the culture period (1485 fold expansion of CIK cells). CIK cells generated from CB showed strong cytotoxic activity against a variety of tumor target cell lines including B and T lymphomas and myeloid leukemias (42–72% killing at a 30:1 E:T ratio). More importantly, they were cytotoxic against AML blasts isolated from 2 patients (41% lysis). During expansion CB derived CIK cells upregulated the NKG2D marker from a mean fluorescence intensity of 49 to 209. Furthermore they expressed perforin and granzyme molecules in &gt;90% of cells. These observations open up the possibility of a future clinical application of this protocol, performed in GMP conditions. Patients relapsing following cord blood transplantation may be treated with CIK cells expanded from the same cord blood unit, where donors would not be anymore available for cell mediated immunotherapy.
Dynamics of BCR-ABL Kinase Domain Mutations in Patients with Chronic Myeloid Leukemia (CML) after Treatment with One, Two or Three Tyrosine Kinase Inhibitors (TKI).
Authors: Jabbour Elias , Jones Dan , Kantarjian Hagop , O’Brien Susan , Garcia-Manero Guillermo , Giles Francis , Wierda William , Cortes Jorge 
Published: April 2020

Abstract
Dasatinib (D) and nilotinib (N) are potent tyrosine kinase inhibitors (TKIs) with activity against many imatinib (IM) resistant BCR-ABL kinase domain mutants, except T315I. In vitro mutant models have selected specific mutations occurring after incubation with IM, D and N. Therapy with these new TKI may select for patients with T315I or other mutations relatively insensitive to them. We assessed the change in mutation status of the bcr-abl kinase domain (codons 220 to 500) in 113 patients (pts) with CML who received therapy with D and/or N after imatinib failure. Median age was 60 years (range, 21 to 82 years). Seventy-one (63%) pts received prior interferon (IFN). Median time on imatinib was 28 months (range, 2 to 78 months). At the time of imatinib failure, mutations were detected in 46 of 85 (54%) pts who had DNA sequencing. The evolution of mutations after a second TKI was as follows (Table 1).
Twenty pts received a third TKI after failing IM and a second TKI. The evolution of mutations in this cohort was as follows (Table 2).
Overall, 19 of 101 evaluable pts (19%), cases had new mutations emerge following TKI switch 17 after a 2nd TKI (12 nilotinib, 5 dasatinib), and 2 after a 3rd TKI (2 dasatinib). We analyzed whether these N- and D-associated new mutations were at sites that have been detected following D and N treatment in vitro (Burgess et al, PNAS 2005; Bradeen et al, Blood 2006; Von Bubnoff et al, Blood 2006). Only 14/46 (30%) kinase domain mutations that developed after D (7) or N (7) corresponded with an in vitro-identified site. Only 5 of 134 (4%) mutations identified were T315I (3 after dasatinib, 2 after nilotinib), but the mutation status of these patients was unknown after IM. We conclude that the spectrum of mutations that develops in vivo after TKI switch is broader and includes common imatinib-resistance sites as well. There appears to no marked increase in the incidence of T315I mutation after TKI switch.
Table 1. Dynamics of mutations after 2nd TKI Post IM mutation No. Post-2nd TKI Mutation (New + Same + Lost) *1 pt acquired new mutation with persistence of pre-existing mutation, 1 lost 3 mutations and acquired 1, and 1 pt lost 2 mutations. Nilotinib Dasatinib Absent 39 8+NA+NA/21 3+NA+NA/18 Present 46 3+20+3/26 2+16+2*/20 Unknown 28 8/9 13/19
Table 2. Dynamics of mutations after 3rd TKI Post IM mutation No. Post-3nd TKI Mutation (compared to status after 2nd TKI) (New + Same + Lost) Nilotinib Dasatinib Absent 5 0/1 1+NA+NA/4 Present 12 0+1+0/1 2+6+3/11 Unknown 3 1/3 NA
The Racial Disparity Gap in Pediatric Leukemia and Lymphoma Survival Has Been Eliminated in Children but Not in Older Adolescents and Young Adults.
Authors: Viny Aaron , Hilden Joanne M. , Stallion Anthony , Bleyer W. Archie 
Published: April 2020

Abstract
Overcoming disparities in the successful treatment of minority cancer patients has been a major challenge. In particular, blacks/African Americans face a higher likelihood of developing and dying from each of the four most common malignancies in the United States (breast, prostate, colon, and lung cancer) as well as from other cancers. Among children, leukemia and lymphoma have also shown a worse outcome in black patients versus whites. Given the success of the Children’s Oncology Group (COG) in reducing the racial deficit in leukemia and lymphoma, we examined national survival data from the U.S. Surveillance, Epidemiology and End Results (SEER) program for trends in survival difference between white and blacks with leukemia and lymphoma, as a function of age at diagnosis. The annual 5-year relative survival rate trends during 1975 to 2000 for all leukemia and all lymphoma (including Hodgkin lymphoma) showed convergence in white and black children (age &lt;10), comparable trends in 10- to 19-year-olds, and divergence in those 20 to 29 years of age with blacks doing progressively worse than whites. The age dependent patterns were qualitatively similar for acute lymphoblastic leukemia, acute myelogenous leukemia, and non-Hodgkin lymphoma, with convergence in &lt;10 year-olds and divergence in 20- to 29-year-olds apparent in each type of leukemia and lymphoma. For the year 2000, the trends in 20- to 29-year-olds project a 20 to 25% survival deficit for blacks versus whites for acute myelogenous leukemia and a 5 to 10% deficit for other leukemia and lymphomas. The fact that the deficits between white and black patients with leukemia and lymphoma have been overcome for pediatric patients in the United States indicates that this can also be achieved in young adults and older adolescents. A major solution would be access to clinical trials through COG or other clinical trials for the next older age group. The current National Cancer Institute Progress Review Group offers recommendations that are relevant to these disparate outcomes observed in this study; these will be reviewed.
Figure Figure
RUNX1/AML1 DNA Binding Domain and ETO/MTG8 NHR2 Dimerization Domain Are Critical to AML1−ETO9a Leukemogenesis.
Authors: Yan Ming , Hiebert Scott , Zhang Dong-Er 
Published: April 2020

Abstract
The 8;21 translocation, which involves the gene encoding the RUNX family DNA binding transcription factor AML1 (RUNX1) on chromosome 21 and the ETO (MTG8) gene on chromosome 8, generates AML1−ETO fusion proteins. Previous analyses have demonstrated that full length AML1−ETO blocks AML1 function and requires additional mutagenic events to promote leukemia in mice. More recently, we have identified an alternatively spliced form of AML1−ETO, AML1−ETO9a, from t(8;21) AML patient samples (Yan et al. Nat. Med.12:945–949, 2006). AML1−ETO9a lacks the C−terminal NHR3 and NHR4 domains of AML1−ETO and is highly leukemogenic in mice. Here, we report that the AML1 DNA binding domain and the ETO NHR2 dimerization domain, but not the ETO NHR1 domain are critical for the induction of acute myeloid leukemia by AML1−ETO9a. Using retroviral mediated gene expression and hematopoietic cell transplantation in recipient mice, we examined AML1−ETO9a, AML1−ETO9a without the NHR1 domain [AML1−ETO9a (dNHR1)] or the NHR2 domain [AML1−ETO9a(dNHR2)], without a histone deacetylase/Sin3A interacting domain between NHR1 and NHR2 [AML1−ETO9a(d350–428)], and mutant AML1−ETO9a proteins that have lost the capacity to bind DNA [AML1−ETO9a(L148D)] and [AML1−ETO9a(R173Q)] in leukemogenesis. All of the mice transplanted with AML1−ETO9a (n =11) and AML1−ETO9a(dNHR1) (n = 12) expressing cells developed acute myeloid leukemia with a similar phenotype (Lin−/c−kit+) within 21 weeks. The median survival times of mice with AML1−ETO9a and AML1−ETO9a(dNHR1) are 9.4 weeks and 10.5 weeks, respectively. Furthermore, all of the mice expressing AML1−ETO9a(d350–428) (n = 11) also developed leukemia with a median survival time of 17.2 weeks. Significant numbers of AML1−ETO9a(d350–428) expressing cells are positive for myeloid markers CD11b and Gr1 in these leukemic mice. In contrast, none of the mice with AML1−ETO9a(dNHR2) (n = 14), AML1−ETO9a(L148D) (n = 8), and AML1−ETO9a(R173Q) (n = 8) expressing hematopoietic cells developed leukemia. Taken together, these data suggest that the AML1 DNA binding domain and the ETO NHR2 domain are required for AML1−ETO9a induced leukemia development and the region between amino acids 350 and 428 of AML1−ETO9a also affects the differentiation stage and latency of leukemogenesis.