bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genome-Wide Mapping of Binding Sites and Networks of Potential Target Genes of the Fusion Oncogene ETV6/RUNX1 in Acute Lymphoblastic Leukemia in Childhood
Authors: Kaulfuss Katja , Heiden Thomas , Hecht Jochen , Seeger Karl 
Published: October 2019

Abstract
Acute lymphoblastic leukemia (ALL) in childhood, a clinically and biologically heterogeneous disease, represents the most common malignant disease in childhood. Approximately 20-25% of B-cell precursor ALL (BCP-ALL) carry the cryptic chromosomal translocation t(12;21)(p13;q22), the most common reciprocal chromosomal translocation in childhood ALL. This translocation combines two transcription factors and essential regulators of normal hematopoiesis, ETV6 and RUNX1, into the fusion oncogene ETV6/RUNX1 (E/R; synonym TEL/AML1).
Recent studies in various animal models have strengthened the view that E/R positive cells give rise to preleukemic clones with a differentiation block in the pro/pre-B stage of B cell development that, after acquisition of additional mutations, may transform into full malignancy.
Regarding the molecular mechanism by which the chimeric fusion protein E/R causes gene expression changes, it is assumed that E/R binds with the runt homology domain of RUNX1 (RHD, DNA-binding domain) to RUNX1 target sequences of gene promoters and recruits corepressors and histone deacetylases through its ETV6 portion, leading to chromatin condensation and transcriptional repression. Thus, E/R appears to act mainly as an epigenetic repressor of genes that are normally activated by RUNX1. However, the precise mechanism of cellular transformation and the identity of E/R target genes are largely unknown.
Therefore, we used chromatin immunoprecipitation (ChIP), followed by next generation sequencing (ChIP-Seq) to identify E/R target genes in the E/R positive BCP-ALL cell lines REH and UoC-B6 as well as in primary patient material from children with relapsed E/R positive ALL. We were able to detect a core gene set of 335 candidate target genes common to all samples analyzed. Those genes could be assigned to 15 significantly overrepresented KEGG pathways (e.g. cell cycle, pathways in cancer, hematopoietic cell lineage and B cell receptor signaling pathway).
The results show, besides target genes already reported in the literature such as EPOR, MPO and IGLL1, numerous not previously described candidate E/R target genes, such as LEF1, E2F2, FLT3, FGFR1 and RUNX1 that are potentially important in the pathogenesis of E/R positive ALL and may lead to new treatment options.

Disclosures
No relevant conflicts of interest to declare.
Nanoparticle-Encapsulated Allogeneic T Cells Mitigate Graft-Versus-Host Disease but Retain Graft-Versus-Leukemia Activity
Authors: Zhang Lingling , Zhao Shuting , Devine Steven M. , He Xiaoming , Yu Jianhua 
Published: October 2019

Abstract
Allogeneic hematopoietic stem cell transplantation (HSCT) has curative potential for hematological malignancies, but is often associated with life-threatening complications including graft-versus-host disease (GVHD). The graft-versus-leukemia (GVL) activity which accompanies HSCT is responsible for eradication of tumor cells and prevention of relapse. GVHD and GVL are usually associated with each other and the separation of the two activities occurs in limited circumstances. In this study, we aimed to mitigate GVHD but retain GVL through transplantation of allogeneic T cells encapsulated with bio-degradable nanoparticle materials.
For the above purpose, donor T cells were encapsulated with chitosan and alginate through layer-by-layer coating using electrostatic deposition. Encapsulated donor T cells were characterized in vitro, and their ability to inhibit GVHD and retain GVL was determined in vivo after being transplanted together with non-encapsulated donor bone marrow (BM) cells in a C57BL/6 → BALB/c HSCT mouse model.
We found 85.7% of donor T cells were successfully encapsulated by the above method (Fig 1A). In vitro studies showed that the encapsulation did not change the phenotype of T cells as defined through the following parameters: size, viability, proliferation, antibody binding, cytokine secretion, and cytotoxicity of T cells (Fig. 1B and data not shown). Mice transplanted with encapsulated allogeneic T cells exhibited less severe acute GVHD and prolonged survival (Fig. 1 C-E). The mice showed a lower GVHD score, less liver damage, a smaller CD8/CD4 T cell ratio, and a higher number of donor BM-derived cells following transplantation with encapsulated donor T cells (Fig. 1 C-E and data not shown). When this GVHD model was combined with implantation of A20 lymphoma cells, GVL of encapsulated T cells was not compromised, while GVHD was still suppressed and the mouse survival also prolonged (Figure 2).
In summary, nanoencapsulation of T cells with bio-degradable materials attenuated the severity of GVHD but retained GVL, presenting a novel and potentially safer and effective approach of allogeneic HSCT for future clinical application.
Figure 1 Figure 1. Figure 2 Figure 2.

Disclosures
No relevant conflicts of interest to declare.
T-Cell Therapy for Multiple Myeloma Using NY-ESO-1+ T-Cell Antigen Presenting Cells (T-APC) Combined with Adoptive Cellular Transfer (ACT) to Augment Immunotherapy
Authors: Patel Krina K. , Olivares Simon , Hurton Lenka V. , Huls Helen , Qazilbash Muzaffar H. , Champlin Richard E. , Cooper Laurence JN 
Published: October 2019

Abstract
T cells have been previously genetically manipulated to act as a cellular vaccine to present viral antigens such as MP1 from influenza A. We have now genetically modified T cells as antigen presenting cells (T-APC) to express the tumor-associated antigen (TAA) NY-ESO-1 (Figure 1). This TAA is reportedly found in the majority of patients with high risk multiple myeloma, as well as other malignancies, but expression is absent from normal tissues. Therefore, immunologic targeting of NY-ESO-1 may be a potential treatment strategy for myeloma that is resistant to conventional therapies. One approach to improve therapeutic outcome is by adoptive transfer of T cells rendered specific for a TAA preferentially expressed on tumor cells. T-cell specificity can be redirected to intracellular TAAs by enforced expression of a characterized T-cell receptor (TCR) or chimeric antigen receptor (CAR) that recognizes processed antigen in the context of restricting human leukocyte antigen (HLA). However, persistence of these infused genetically modified cells, which is directly related to a favorable clinical response, may be variable. Therefore, T-APC may have a dual function of increasing persistence of adoptively transferred CAR+ and/or TCR+ T cells as well as inducing long term immunity through direct and cross priming of endogenous immune cells. To enhance these presentation functions, a new co-stimulatory molecule that tethers IL-15 to the cell surface (membrane-bound IL-15, mIL15) was expressed on NY-ESO-1+T-APC using the Sleeping Beauty (SB) gene transfer system. NY-ESO-1-specific HLA A2+ T cells expressing TCR and CAR could be selectively propagated ex vivo on autologous mIL15+NY-ESO-1+T-APC. The T-APC were also able to selectively propagate NY-ESO-1-specific T cells from autologous peripheral blood which can be described by binding of specific pentamer (Figure 2). Our data demonstrate that it is feasible to generate T-APC that can activate T cells engineered to have a defined specificity for NY-ESO-1 as well as grow out NY-ESO-1-specific T cells. The SB system is in place for genetic modification of clinical-grade T cells to express CAR, TCR, NY-ESO-1, and mIL15. Thus, we are proceeding to a human trial to infuse T-APC with and without genetically modified T cells in patients with NY-ESO-1+ tumors such as myeloma.
Figure 1 Figure 1.
Figure 2 Figure 2.

Disclosures
Cooper: InCellerate: Equity Ownership; Sangamo: Patents & Royalties; Targazyme: Consultancy; GE Healthcare: Consultancy; Ferring Pharmaceuticals: Consultancy; Fate Therapeutics: Consultancy; Janssen Pharma: Consultancy; BMS: Consultancy; Miltenyi: Honoraria.
Relationship between Gene Polymorphism of Recipient/ Donor Immune Pathway and Invasive Fungal Infection after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Shi Jimin , Xiao Haowen , Luo Yi , Tan Yamin , He Jingsong , Xie Wanzhuo , Ye Xiujin , Cai Zhen , Lin Maofang , Huang He 
Published: October 2019

Abstract

Invasive fungal infections are a major cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation(allo-HSCT). During the past 2 decades, changes in transplantation practices and strategies to diagnose and treat IFI have likely impacted the epidemiology of IFI. Polymorphism of genes, which take part in the allogeneic immune response after allo-HSCT, lead to allograft immunoreactivity differences between individuals. It also may affect IFI after transplantation.
Methods: We analyzed allo-HSCT recipients at our center during the period January 2001 through March 2009. We analysis gene polymorphism of innate immune gene TLRs among 240 pairs of specimens of recipients and donors, in order to find out IFI individual difference and high risk genotype of IFI after transplantation.
Results: There were 99 patients (41.2%) occured IFI after allo-HSCT. Hematological malignant diseases, serious acute GVHD, and extensive chronic GVHD were risk factor for IFI after allo-HSCT. Univariate analysis of the relationship between risk factors of IFI after all0-HSCT and the TLRs gene polymorphism, we found that in the 10 SNP loci in 5 TLRs , two polymorphisms of TLR8 gene, (+1 A/G, rs3764880; +354 C/T, rs2159377) effect the incidence of IFI after allo-HSCT. Other gene polymorphism of TLRs (TLR1, TLR2, TLR3, TLR9) had no significant effects on the risk of IFI after allo-HSCT.
Conclusion: The study of the relationship between the gene polymorphism of natural immune molecule TLRs and the risk factors after allo-HSCT showed that TLR8 genotype of donor stem cell significantly influenced the incidence of IFI after allo-HSCT at the first time. It provide the evidence to establishment a fungal infection index system, which based on risk stratification of genetic background before transplantation of donors and recipients stem cell, and provide basis for selection of unrelated donor and prevention and treatment of IFI

Disclosures
No relevant conflicts of interest to declare.
Research on the Key T Cells and Cytokines of aGVHD after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Tan Yamin , Luo Yi , Zhao Yanmin , Liao Weichao , Fu Huarui , Shi Jimin , Cai Zhen , Huang He 
Published: October 2019

Abstract
Objective: Acute graft versus host disease(aGVHD) is due to one of the most important causes of treatment related mortality after allo-HSCT. T cells play the key roles in the development of aGVHD after allo-HSCT, and the different subsets play different roles. Research on the reconstruction of T cell immune function and its cytokines in the occurrence of aGVHD has become one of the hottest topic in aGVHD research. We investigated the key T lymphocyte subsets and its cytokines, to explore the relationship between the characteristics of T cell subsets and the process of aGVHD. We desire to find some biomarkers and establish early diagnosis index for aGVHD, furthermore find an important target for immunotherapy. Ultimately the strategy could reduce aGVHD related mortality and improve long-term disease-free survival in transplantation patients.
Methods: 64 patients were included in the study, from May 2013 to January 2014. Blood samples of these patients were monitoring every week. Treg, Th1 and Th17 cells number were detected by flow cytometry. Serum levels of IL-2, IL-6, IL-10, IL-17, TNF-a and IFN-γwere detected by flow cytometry (CBA). ELISA technique was used for the detection of serum TGF- β level.
Results: 36 patients (56.3%) suffered from II-IV aGVHD, and 10 cases (15.6%) among them was diagnosed of III°-IV° aGVHD. Before the onset of aGVHD, Treg, Th1, Th17 cells levels had no statistical difference between aGVHD patients and non aGVHD patients. In aGVHD patient, serum levels of IL-2 (P=0.0417) and TGF- β (P=0.0168) elevated in the early stage of aGVHD. IL-2 (P=0.0231), IL-10 (P=0.0089),TGF-β (P=0.0138) serum levels after aGVHD was significantly higher than that in non aGVHD patients, and Treg cellsnumber was significantly lower than that of patients without aGVHD (P=0.0150). Using ROC and logistic regression analysis, we found a composite biomarker panel (IL-2, IL-10 and Treg cell, which could optimally discriminated patients with and without aGVHD, the formula is: 1.656+0.103×IL-2|0.052 ×IL-10|0.037×Treg. ROC curve of composite panel is 0.89 (95% CI, 0.78-0.99, P=0.0002), which is larger than that of any single index.
Conclusion: Increased IL-2, IL-10, TGF-β serum levels in early stage after allo-HSCT can help to establish the clinical diagnosis of aGVHD, and which is expected to provide a clinical basis for further target immunotherapy. Treg cell level was negatively related to the onset of aGVHD. Furthermore, we found a composite biomarker panel (IL-2, IL-10 and Treg cell), which could optimally discriminated patients with and without aGVHD. It could provide a reliable basis for early diagnosis of aGVHD.

Disclosures
No relevant conflicts of interest to declare.
Donor Cell Leukemia: A Prospective Study of Its Identification and Treatment
Authors: Ruiz-Delgado Guillermo J , Gonzalez-Ramirez Monica P. , Martagon-Herrera Nora A , Garces-Eisele Javier , Ruiz-Arguelles Alejandro , Gonzalez-Cortes Angelica , Ruiz-Arguelles Guillermo J. 
Published: October 2019

Abstract
Leukemia relapses occurring in donor cells, so called donor cell leukemias (DCL) after allogeneic hematopoietic stem cell transplantation have been reported in several cases and still are considered as rare diseases. Cytogenetic analysis, flow cytometry and molecular testing have been used to confirm this event in the cases so far reported. The etiology of DCL is unclear, and the reported literature does not suggest a common mechanism. The incidence of this condition is largely unknown, as well as the results of its treatment. We have prospectively searched for DCL in a 12-year period, in a single institution. In a group of 106 consecutive patients allografted because of leukemia; we have identified 7 cases of DCL; six of them were allografted because of relapsed acute lymphoblastic leukemia (ALL) and one because of paroxysmal nocturnal hemoglobinuria / aplastic anemia; these figures suggest that the real incidence of DCL has been underestimated in previous studies. All the patients were allografted from HLA-identical siblings, employing a reduced-intensity conditioning regimen. The cases of DCL appeared one to 40 months (median 10) after the allograft; the number of blast cells when the leukemic activity ensued was above 50% in all cases, whereas the chimerism studies revealed more than 90% cells of donor origin. The origin of the leukemia cells was shown by microsatellites in all cases and in three with a sex mismatch it was confirmed by the enumeration of XX / XY cells in the leukemic cells. The six patients with lymphoblastic DCL were treated prospectively with a pediatric-inspired combined chemotherapy schedule designed for “de novo” ALL patients. A complete response was obtained in 3/6 patients with lymphoblastic DCL these patients being alive in a complete remission at 11, 12 and 98 months after the diagnosis of DCL. The patient with hairy cell DCL has had a benign course, not needing any treatment. In the whole group the median survival (SV) has not been reached, the overall SV being 57% at 98 months. The long-term DCL survivors remain full chimeras and did not need a second transplant. It is concluded that the prevalence of DCL may be higher if it is prospectively looked for, and that acceptable therapeutic results are obtained if patients are treated as “de novo” leukemias employing combined chemotherapy.

Disclosures
No relevant conflicts of interest to declare.
Chemotherapy Versus Hypomethylating Agents for the Treatment of Relapsed Acute Myeloid Leukemia and Myelodysplastic Syndrome Following Allogeneic Stem Cell Transplant: A Retrospective Review
Authors: Motabi Ibraheem H , Liu Jingxia , Abboud Camille N. , Cashen Amanda F. , Stockler-Goldstein Keith E. , Uy Geoffrey L. , Vij Ravi , Westervelt Peter , DiPersio John F. 
Published: October 2019

Abstract
Allogeneic stem cell transplant (allo-SCT) is a potentially curative option for high risk acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). Disease relapse after transplant is the major cause of treatment failure and is associated with a poor outcome. Intensive chemotherapy followed by donor lymphocytes infusion (DLI) or a second SCT may result in a durable response in some patients, but is associated with increased toxicity. More recently, a less aggressive therapy with hypomethylating agents (HA) has been reported to have activity in treatment of post-SCT relapse. We compared the treatment outcomes of intensive chemotherapy with that of HA. The primary end points were overall response rate (ORR) and overall survival (OS). Secondary endpoints were complete remission (CR) rate and progression free survival (PFS). A total of 100 patients with AML in morphological evidence of relapse were included: 73 patients received chemotherapy and 27 patients received a HA. Fifty-six percent of patients in the chemotherapy group and 33% of patients in the HA group received at least one DLI after treatment. Treatment with chemotherapy resulted in a higher ORR (51% vs. 18.5%, p = 0.004) and a higher CR rate (40% vs. 7%, p= 0.002). The median OS (6 months vs. 4 months, p = 0.024) and PFS (8 months vs. 2.3 months, p = 0.053) were longer in chemotherapy group. The overall survival (OS) at one year was 32% in the chemotherapy group compared with 4% in the HA group (p = 0.024). Similar benefit of chemotherapy over HA was maintained in all treatment outcomes after controlling for the use of DLI. The use of chemotherapy followed by DLI may offer the greatest benefit ( ORR of 68%; 1-year OS of 44%; and median OS of 18 months). Conclusion: our results suggest that intensive chemotherapy may result in a better response rate and a better overall survival compared with hypomethylating therapy in patients with morphological relapse of AML after SCT. We propose prospective studies before the use of hypomethylating agents in this setting for medically fit patients.

Disclosures
Off Label Use: Rituximab is a genetically engineered monoclonal antibody that targets a specific protein, known as CD20 found on the surface of normal and malignant B-lymphocytes.Use of Rituximab treatment for other than FDA approved indications include a rare condition of acquired factor VIII inhibitors in individuals without inherited hemophilia as an autoimmune phenomenon that may lead to life-threatening bleeding. Due to the low incidence rate of acquired inhibitors, published data consists of only case reports, and reviews.. Vij:celgene: Honoraria, Research Funding.
Cpeb4-Mediated Translational Regulatory Circuitry Controls Terminal Erythroid Differentiation
Authors: Hu Wenqian , Yuan Bingbing , Lodish Harvey F. 
Published: October 2019

Abstract
While we have considerable understanding of the transcriptional networks controlling mammalian cell differentiation, our knowledge of post-transcriptional regulatory events is very limited. Using differentiation of primary erythroid cells as a model, we show that the sequence-specific mRNA-binding protein Cpeb4 is induced by the erythroid important transcription factors Gata1 and Tal1, is strongly upregulated during terminal erythroid development, and is essential for terminal erythropoiesis. By binding directly to the translation initiation factor eIF3 complex, Cpeb4 represses the translation of a large set of mRNAs, most of which are normally downregulated during terminal erythroid development. Cpeb4 also binds to its own mRNA to repress its translation, and ectopic expression of Cpeb4 blocks erythroid differentiation. Thus transcriptional induction and translational repression combine to form a negative feedback loop to control Cpeb4 protein levels within a specific range that is required for terminal erythropoiesis. Our study provides an example of how translational control is integrated with transcriptional regulation to precisely control gene expression during mammalian cell differentiation.

Disclosures
No relevant conflicts of interest to declare.
Dietary Supplementation with Ipriflavone Decreases Liver Iron Loading in Wild Type Mice
Authors: Patchen Bonnie , Gaun Vera , Cheng Aaron , Fraenkel Paula G. 
Published: October 2019

Abstract
Hepcidin, a peptide hormone produced in the liver, decreases intestinal iron absorption and macrophage iron release by causing internalization and degradation of the iron exporter ferroportin. Because its levels are inappropriately low in patients with iron overload syndromes, Hepcidin is a potential drug target. We previously conducted a chemical screen in human hepatocytes (HepG2 cells) to identify small molecules that upregulate Hepcidin transcript levels. One of the small molecules that we identified was ipriflavone, a synthetic isoflavone, which has been used to treat osteoporosis in postmenopausal women. To evaluate ipriflavone’s effect on iron homeostasis in a mammalian model, we placed groups of 5-week old C57BL/6 male mice on a soy-free, iron-sufficient diet, AIN-93G containing 220 mg/kg iron and 0, 250, 500 or 750 mg ipriflavone per kg of food (n=4 or 5 per group) for 50 days, then sacrificed the animals for analysis of changes in gene expression by quantitative realtime RT-PCR, liver and spleen iron content, blood indices, and intestinal ferroportin expression. While producing less than a two-fold increase in liver hepcidin transcript levels, ipriflavone supplementation was associated with a significant decrease in liver iron content (mean±SE): 39.22±2.06 µg iron/g tissue, p=0.0033, and 44.10±2.58 µg iron/g tissue, p=0.0124, at 500 and 750 mg ipriflavone per kg of food, respectively, vs 57.77±3.39 µg iron/g tissue in mice that did not receive ipriflavone. Ferroportin expression detected in intestinal epithelial cells by immunohistochemistry was notably decreased in mice receiving ipriflavone: 66.6% and 80% of the animals receiving 500 and 750 mg/kg groups, respectively, exhibited decreased ferroportin staining versus 40% of the group not receiving ipriflavone. There was no significant difference in hemoglobin, hematocrit, or spleen iron among the groups. In conclusion, we have shown that orally administered ipriflavone is effective in decreasing liver iron content and intestinal ferroportin expression in vivo. Future experiments will evaluate ipriflavone’s effects on iron homeostasis and hematopoiesis in genetic models of iron overload disorders.

Disclosures
No relevant conflicts of interest to declare.