bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rituximab for Desensitization during HLA-Mismatched Stem Cell Transplantation in Patients with a Positive Donor Specific Anti-HLA Antibody: A Prospective Study
Authors: Chang Ying-Jun , Xu Lan-Ping , Wang Yu , Zhang Xiao-Hui , Chen Huan , Chen Yu-Hong , Wang Feng-Rong , Han Wei , Sun Yu-Qian , Yan Chenhua , Tang Fei-Fei , Huo Ming-Rui , Mo Xiao-Dong , Zhao Xiang-Yu , Liu Kai-yan , Huang Xiao-Jun 
Published: November 2019

A prospective, clinical trial in non-manipulated haploidentical allografts was initiated to define the efficacy of a single dose of 375 mg/m2 rituximab as a desensitization regimen for donor-specific anti-HLA antibody (DSA)-positive patients with 2,000 ≤ mean fluorescence intensity (MFI) < 10,000. In the clinical cohort, compared to pretransplantation [median, 4791, n=28], the median MFI levels at day 7 [0], 14 [0], 21 [0], 30 [0], and 45 [0] following transplantation were significantly decreased (P<0.0001 for all). The incidence of primary poor graft function (PGF) in the clinical cohort (n=55) was comparable to that in the matched-pair cohort (n=110) negative for the DSA (5% vs. 1%, P=0.076), both of which were significantly lower than that in the historical cohort (n=22) with 2,000 ≤ MFI<10,000 without receiving any treatment to cope with DSA (27%, P<0.001, for all). Rituximab desensitization was associated with a reduced incidence of primary PGF (HR 0.200, P=0.023). The 3-year non-relapse mortality of patients in the clinical cohort and the matched-pair cohort were 23% and 24%, respectively, both of which were lower than that of patients in the historical cohort (37%), although no statistical significance was observed. These results led to a low 3-year overall survival in patients in the historical cohort (58%) compared to those in the clinical cohort (71%) and the matched-pair cohort (73%). These results suggest that a single dose of rituximab could be effectively used to desensitize patients and prevent the onset of primary PGF in patients who receive HLA-mismatched allografts.
This study is registered at http://www.chictr.org.cn/ChiCTR-OPC-15006672.

Disclosures
No relevant conflicts of interest to declare.
Deregulated CXCR4-CXCL12 Signaling Impacts on the Pathogenesis of DLBCL
Authors: Pansy Katrin , Feichtinger Julia , Ehall Barbara , Uhl Barbara , Greinix Hildegard T. , Prochazka Katharina , Beham-Schmid Christine , Neumeister Peter , Wrodnigg Tanja , Fechter Karoline , Deutsch Alexander 
Published: November 2019

Introduction:
The interaction of the chemokine receptor CXCR4 and its ligand CXCL12 appears to be implicated in many important biological processes such as proliferation, survival, migration, and/or invasion. Furthermore, it is important for normal leukocyte trafficking. Deregulation of this axis is frequently observed in several hematologic malignancies. In diffuse large B cell lymphomas (DLBCL), the CXCR4-CXCL12 axis is still largely unexplored and published data are contradictive. Hence, we comprehensively studied the CXCR4-CXCL12-axis in our DLBCL cohort as well as the effects of CXCR4 antagonists on lymphoma cell lines in vitro.
Methods:
We determined the CXCR4 and CXCL12 expression levels in NGCB- and GCB-DLBCL consisting of primary and transformed follicular lymphomas (n=77 in total), the corresponding bone marrow samples (n=63) and non-neoplastic germinal center-B cells (GC-B, n=5) serving as non-malignant control. To investigate the effects of CXCR4 antagonists in vitro and their function in regulation of important pathways (JNK, ERK1/2 and NF-κB) known to be involved in lymphomagenesis, we treated lymphoma cell lines with three different CXCR4 antagonists, AMD070, AMD3100 and WK1 (a novel nicotinic acid derivative of AMD070 - synthesized by us), followed by functional assays and gene expression profile.
Results:
CXCR4 was 140-fold higher expressed in DLBCL compared to non-neoplastic GC-B cells. Interestingly, higher CXCR4 expression correlated to a clinically advanced stage, to bone marrow infiltration and worse cancer-specific survival in DLBCL. Further expression analysis by using the corresponding bone marrow biopsies demonstrated that CXCL12 expression correlated to the lymphoma infiltration rate and that CXCR4 expression was reduced in remission under therapy. Moreover, two CXCR4 antagonists - AMD070 and especially WK1 - exerted pro-apoptotic effects on CXCR4 positive lymphoma cells in vitro and induced the expression of certain pro-apoptotic genes in CXCR4 positive cell lines. Remarkably, these effects were more pronounced for the WK1. Finally, WK1 treatment resulted in reduced expression of JNK-, ERK1/2- and NFκB/BCR-target genes.
Conclusion:
Our data demonstrate that the CXCR4-CXCL12 axis is involved in the pathogenesis of DLBCL. Since CXCR4 antagonists exert pro-apoptotic effects and impact lymphoma relevant pathways, they represent interesting molecules to develop novel therapeutic agents.

Disclosures
No relevant conflicts of interest to declare.
SIX1 Is a Novel Effector Gene in CALM-AF10 Leukemia
Authors: Aumann Waitman K. , Lavau Catherine P. , Harrington Amanda , Conway Amanda E. , Wechsler Daniel S. 
Published: November 2019

Background: The CALM-AF10 translocation is detected in ~10% of T-cell acute lymphoblastic leukemias (T-ALLs), and in some acute myeloid leukemias (AMLs). CALM-AF10 leukemias are characterized by high expression of proleukemic HOXA genes, which serve a critical role in hematopoiesis. We hypothesized that identification of novel CALM-AF10 effector genes may yield new therapeutic targets in this difficult to treat leukemia. We took advantage of our prior observation that the nuclear export factor CRM1/XPO1 tethers CALM-AF10 to HOXA genes by interacting with a nuclear export signal (NES) in CALM. Using next generation sequencing, we determined that, SIX1, similar to HOXA genes, is increased in CALM-AF10 leukemias and decreased in response to CRM1 inhibition with Leptomycin B (LMB). Design/Methods: RT-qPCR and Chromatin Immunoprecipitation were performed using both bone marrow progenitors and murine embryonic fibroblasts (MEFs) transduced with CALM-AF10 or an empty vector, with and without LMB. The ability of SIX1 to enhance self-renewal of hematopoietic progenitors was examined by measuring the colony-forming ability of transduced fetal liver hematopoietic progenitor cells. CRISPR-Cas9 was used to silence SIX1 in Human Embryonic Kidney 293 (HEK293) cells. Results: RT-qPCR confirmed overexpression of SIX1 in both CALM-AF10 transduced MEFs and CALM-AF10 leukemias, with decreased SIX1 expression observed in the presence of LMB. ChIP analysis showed that CALM-AF10 binds to the SIX1 gene locus. Overexpression of SIX1 in fetal liver cells was sufficient to increase the self-renewal potential of colony-forming progenitors. SIX1 was successfully knocked out in HEK293 cells without a significant effect on HEK293 proliferation. Conclusions: The SIX1 homeobox gene is highly expressed during development and its expression is silenced post-embryogenesis. Increased SIX1 expression has been reported in numerous solid tumors. We have determined that SIX1 is upregulated in CALM-AF10 leukemias, and increases the self-renewal potential of hematopoietic progenitors. Using CRISPR-Cas9 to silence SIX1, we have demonstrated that SIX1 is not essential for cell survival, and that its inhibition may impair CALM-AF10 leukemia cell proliferation. Thus, SIX1 may play a pathogenic role in leukemogenesis and is a potential therapeutic target in CALM-AF10 leukemias.

Disclosures
No relevant conflicts of interest to declare.
Impact of Iron Overload and Iron Removal Therapy on Outcome of Allogeneic Hematopoietic Stem Cell Transplantation in the Patients with Severe Aplastic Anemia
Authors: Pan Tianzhong , Tang Baolin , Zhu Xiaoyu , Liu Huilan , Song Kaidi , Wan Xiang , Yao Wen , Sun Zimin 
Published: November 2019

Abstract:The aim of this study was to evaluate the Impact of iron overload and iron removal therapy on the outcome of allogeneic hematopoietic stem cell transplantation (SCT) in patients with severe aplastic anemia (SAA). Forty-five transplant patients with SAA were retrospectively analyzed. Patients were divided into two groups according to whether there was iron overload or not: iron overload group (n=23) and non-iron overload group (n=22). The iron overload group had higher 1-year transplant-related mortality (TRM) (30.4% vs. 4.5%, P = 0.02), lower 1-year transfusion-free survival (TFS) (40.6% vs. 81.8%, P = 0.007) and lower 1-year overall survival (OS) (54.8% vs. 90.2%, P = 0.003) than the non-iron overload group. However, there was no significant difference in implantation rate, pre-engraftment syndrome (PES) and acute graft-versus-host disease (aGVHD, grade III-IV) between the two groups. Furthermore, the multivariate analysis revealed that iron overload was an independent risk factor affecting OS (HR = 36.88, P = 0.03). The patients in iron overload group were divided into two subgroups according to the time of iron removal using deferasirox 20mg/kg/d. Specifically, group A (n=17) was treated with deferasirox 20mg/kg/d for 3 months or less and group B (n=6) treated for more than 3 months. The results showed that group B had a lower one-year TRM (0% vs. 41.2% , P = 0.07), higher 1-year TFS (66.7% vs. 31.4%, P = 0.18) and higher 1-year OS (100% vs. 31.4% , P = 0.02) than group A. There were no significant difference in implantation rate, PES and aGVHD ( III-IV) between the two groups. In conclusion, the results indicate that iron overload before transplantation was associated with increased TRM and decreased OS, suggesting that iron overload was associated with worse transplantation outcomes. However, standardized iron removal therapy could reverse the adverse effects of iron overload on the outcomes of transplantation to a certain extent and improve the prognosis significantly.

Disclosures
No relevant conflicts of interest to declare.
Elevated ANXA1 Expression Causes Glucocorticoid Resistance in Acute Lymphoblastic Leukemia Via Activating the Wnt/β-Catenin Signaling Pathway
Authors: Liu Ping , Ma Dan , Wang Jishi 
Published: November 2019

Background: Acute lymphoblastic leukaemia (ALL) is one of the most common clonal malignant diseases in children, and it stems from unchecked proliferation of lymphoid progenitor cells. Glucocorticoids (GCs) such as prednisolone and dexamethasone are used as a chemotherapeutic drug in the treatment of ALL. GC-induced cell mortality is first mediated by the activation of glucocorticoid receptor (GR), followed by its translocation into the nucleus to activate or inhibit gene transcription. However, up to ~20% patients with leukemia relapse and become resistant to GCs. Therefore, a better understanding the molecular basis of chemoresistance in ALL would provide novel therapeutic opportunities for patients. Methods: By analyzing the published mRNA expression profiles (GSE5280; GSE94302) obtained from NCBI (https://www.ncbi.nlm.nih.gov/geo/), we found that higher expression of ANXA1 was significantly associated with decreased overall survival of ALL patients. We also examined the expression of ANXA1 at mRNA and protein levels in a variety of ALL cell lines by using qRT-PCR and western blot analyses. The mRNA and protein expression of ANXA1 in ALL cell lines and patients were determined using Real-time PCR and Western blot respectively. Functional assays, such as CCK-8, FACS, and Tunel assay used to determine the oncogenic role of ANXA1 in ALL progression. Furthermore, western blotting and luciferase assay were used to determine the mechanism of ANXA1 promotes chemoresistance in ALL cells. Results: The expression of ANXA1 was markedly upregulated in ALL cell lines and patients, and high ANXA1 expression was associated with relapsed/refractory ALL patients. ANXA1 overexpression confers glucocorticoids (GCs) resistance on ALL cells; however, down-regulated of ANXA1 sensitized ALL cell lines to GC both in vitro and in vivo. Additionally, ANXA1 upregulated the levels of FPRs by promoting Wnt/β-catenin signalling. Conclusions: Our findings provided evidence that ANXA1 is a potential therapeutic target for patients with ALL. Targeting ANXA1 signaling may be a promising strategy to enhance GC response during ALL chemo-resistance.

Disclosures
No relevant conflicts of interest to declare.
A Case of Acute Lymphocytic Leukaemia with t(3;13) and Central Nervous System Leukemia after Allogenic Cord Blood Transplantation
Authors: Hong Yaqun , Li Xiaofan , Jiafu Huang , Xu Zhenshu , Li Nainong 
Published: November 2019

Background: Acute lymphoblastic leukemia (ALL) is a neoplastic cancer characterized by clonal expansion of leukemic cells in lymph organs and bone marrow. Lots of kinds of different chromosomal translocation can be found in those leukemic cells. However, the role of the abnormal chromosomals and genes in leukemogenesis is not yet fully understood. Identifying new chromosomal translocation can facilitate a better understanding of pathogenesis of this disease.
Case presentation: We report a rare case of acute lymphocytic leukaemia with t(3;13)(q29,q21). The patient was diagnosed pre-B-ALL with no abnormal chromosomal or gene fusion and achieved complete remission (CR) after induction chemotherapy. Ten months later, she relapsed in the consolidation with cytogenetics test showing 46,XX,t(3;13)(q29,q21). Given no CR after 2 chemotherapy regimens, she received salvage cord blood transplantation. Regular intrathecal methotrexate was applied to prevent central nervous system leukemia. Good graft-versus-leukemia (GVL) effect was induced by daily injection of low dose of IL-2 two months post transplantation. Minimal residual disease (MRD) negativity was maintained until central nervous system leukemia was found 8 months after transplantation. A whole exome sequencing was performed. Nine driver mutation genes and seven tumor genes were found.
Conclusions: We highly suspect that the relapse in central nervous system after transplantation is associated with the rare chromosomal translocation.

Disclosures
No relevant conflicts of interest to declare.
Molecular Classification of Small B-Cell Lymphomas Using Digital Multiplexed Gene Expression Applicable to Formalin-Fixed Paraffin-Embedded Tissues
Authors: Xiao Min , Zhou Jianfeng , Zhang Wei 
Published: November 2019

Background: Small B-cell lymphomas encompass a group of lymphoid neoplasms with high heterogeneity. However, the lack of relatively specific diagnostic markers for most of these diseases make their diagnosis challenging.
Methods: Using NanoString platform, a random forest-based molecular classification model was trained on 203 malignant and 98 non-malignant formalin-fixed paraffin-embedded (FFPE) tissues. Candidate genes were selected from microarray gene expression data of 891 small B-cell lymphomas. All malignant biopsies were derived from individuals diagnosed as a well-defined small B-cell lymphoma entity, including 17 chronic lymphocytic leukemia/small lymphocytic B-cell lymphoma (CLL/SLL), 74 follicular lymphoma (FL), 34 classical mantle cell lymphoma (cMCL), 5 non-nodal mantle cell lymphoma (nnMCL), 56 mucosa-associated lymphoid tissue lymphoma (MALT), 3 splenic marginal zone lymphoma (SMZL), and 14 nodal marginal zone lymphoma (NMZL). A leave-one-out prediction (CLL/SLL, FL, MCL, MZL, and non-malignancy) was applied to test the accuracy of prediction of the model, and Gini Index of each gene was taken into consideration in simplifying the molecular assay applicable to routine test.
Results: A total of 154 candidate genes were initially selected for training the molecular classification model. Genes were initially selected for training the molecular classification model. Thirty-two genes were specifically highly expressed in a single subgroup with Gini Index more than 0.001. The most important genes for classification were CCND1, ZBTB32, MME, BHLHE41, SOX11, and RGS13. The most distinguishable entity was CLL/SLL (ZBTB32, CLNK, and CD200), followed by cMCL/nnMCL (CCND1 and SOX11), and FL (MME, RGS13, and ELL3). Using the molecular classification model trained on the most specific 33 genes, the consistency of diagnosis were 97.5%.
Conclusions: In conclusion, our molecular model robustly recognized different subtypes of small B-cell lymphomas in leukemic FFPE samples, which provides a novel approach to precise diagnosis and classification of small B-cell lymphomas.
Figure

Disclosures
Zhou: Nanjing Iaso Biotherapeutics Co. Ltd.: Other: Chairman of Advisory Committee of Science and Medicine.
The Role of Gα13 in Integrin β2-Dependent Neutrophil Transendothelial Migration
Authors: Chang Claire W , Cheng Ni , Du Xiaoping 
Published: November 2019

Upon sensing chemoattractant signals, leukocytes migrate towards the chemoattractant. If the signal is extravascular, circulating leukocytes roll on the surface of endothelial cells, firmly adhere and migrate through the endothelial monolayer to the inflammation site. Transendothelial migration of leukocytes requires integrins, particularly β2 family of integrins. We previously demonstrated that the heterotrimeric G protein subunit Gα13 interacts with the cytoplasmic domains of certain integrin β subunits including β2. To understand the role of Gα13 in integrin-dependent neutrophil migration, myeloid-specific Gα13 deficient mice have been generated by cross-breeding lysozyme M-cre mice with Gα13-floxed mice. Neutrophil migration was attenuated in myeloid-specific Gα13 knockout mice in a thioglycolate (3%) induced peritonitis model in vivo, and was also attenuated in transendothelial or mouse ICAM-1-coated transwell migration assays ex vivo. In contrast, endothelial cell/ICAM-independent transwell migration was not affected. Importantly, on an ICAM-coated surface, only the speed of migration of Gα13-deficient neutrophils were attenuated but migration directionality was not affected, suggesting that Gα13 plays an important role in motility rather than chemoattractant sensing. To further determine the role of Gα13-β2 integrin interaction in leukocyte migration, we examined the effect of a peptide, MB2mP6, derived from the Gα13 binding site of integrin β2 subunit. Like Gα13 deficiency, MB2mP6 inhibited integrin-dependent neutrophil transendothelial migration, but had minimal effects on leukocyte transwell migration in the absence of endothelial cell monolayer. Gα13 plays dual role in RhoA regulation. GPCR-activated Gα13 binds to and activates RhoGEFs resulting in RhoA activation, but Gα13 binding to integrin cytoplasmic domain is associated inhibition of RhoA. Interestingly, treatment with MB2mP6 enhanced RhoA activity of neutrophils adherent on ICAM1, suggesting that MB2mP6 mainly affects Gα13-β2 interaction. These data suggest that Gα13-integrin interaction plays an important role in the integrin-dependent transendothelial migration. Thus, Gα13-β2 is important in the immune and inflammatory functions of neutrophils.

Disclosures
No relevant conflicts of interest to declare.
High Dimensional Immune Profiling in Smoldering Multiple Myeloma Identifies Novel Organizing Features of the Tumor Microenvironment
Authors: Cho Hearn Jay , Perumal Deepak , Rahman Adeeb H , Kim-Schultze Seunghee , Yesil Jennifer , Auclair Daniel , Casneuf Tineke , Adams Homer , Parekh Samir , Gnjatic Sacha , Fernandez Nicolas 
Published: November 2019

Multiple myeloma (MM) is a malignancy of plasma cells that arises from premalignant Monoclonal Gammopathy of Undetermined Significance (MGUS) and often progresses through an asymptomatic Smoldering (SMM) phase lasting months or years before manifesting clinical symptoms warranting therapy. Current research indicates that the tumor microenvironment (TME) in the bone marrow may play a significant role in governing progression to symptomatic disease. Therefore, understanding of the interactions between malignant plasma cells and the TME in early disease states is critical in the pursuit of therapies that will prevent progression to symptomatic disease. We performed high dimensional genomic and immunologic analysis of bone marrow specimens from 73 subjects with SMM. We performed RNA-seq on the malignant plasma cells isolated by anti-CD138 magnetic bead positive selection, mass cytometry (CyTOF) and T cell receptor sequencing (TCR Seq) of CD138-depleted bone marrow mononuclear cells, and proteomics, seromic, and grand serology analysis of bone marrow plasma. These samples and assays provided a broad view of the tumor cells and the cellular and soluble components of the TME. Each of these assays identified self-organizing clusters of subjects, indicating that subgroups of SMM patients shared common characteristics in the tumor or TME populations. We then applied novel bioinformatic methods to compare data from pairs, trios, quartets, and quintets of assays to identify communities of subjects with similar immunologic and genomic characteristics. Integrated analysis of CyTOF, proteomic, and TCR Seq resolved three distinct communities with a high degree of significance. These communities shared distinct cellular and proteomic features that suggested early adaptive, activated adaptive, or innate immune characteristics. These results suggest that the continuum from MGUS to MM does not consist of a single pathway in either the tumor cells or the TME, and that complex interactions ultimately determine progression. This suite of assays (CyTOF, proteomics, and TCR Seq) may be applicable in translational and clinical studies to understand key tumor and immune determinants of SMM and lead to rationally designed therapy to replicate these conditions to prevent progression to symptomatic disease.

Disclosures
Cho: Genentech: Honoraria, Research Funding; Celgene: Honoraria, Research Funding; The Multiple Myeloma Research Foundation: Employment; Takeda: Research Funding; BMS: Consultancy; Agenus: Research Funding; GSK: Consultancy. Adams:Janssen Pharmaceuticals R&D: Employment, Other: Own Stock. Parekh:Foundation Medicine Inc.: Consultancy; Celgene Corporation: Research Funding; Karyopharm Inc.: Research Funding.
Patients with Bone and Bone Marrow Involvement Had Better OS and PFS in Patients with Aggressive Non-Hodgkin Lymphoma Treated with CD19 CAR T Cells
Authors: Zhou Lili , Aibin Liang , Li Shaoguang , Ye Shiguang , Li Ping 
Published: November 2019

Factors associated with complete remission and durable remission after CD19 chimeric antigen receptor (CAR)-modified T-cell immunotherapy for aggressive B-cell non-Hodgkin lymphoma (NHL) have not been identified. We report multivariable analyses of factors affecting response and progression-free survival (PFS) in patients with aggressive NHL treated with CD19 CAR T cells. The best overall response rate was 71%, with 32% of patients achieving complete remission. The median PFS and OS of patients with aggressive NHL who achieved complete remission were 11 months and 12 months. The median PFS and OS of patients with bone and bone marrow involvement without soft tissue involvement were 14 months and 18 months. The median PFS and OS of patients with soft tissue involvement were 1 months and 4 months. We report complete remission rates and long-term survival rates in patients with bone and bone marrow involvement were significantly higher than those in patients with soft tissue involvement.
Figure

Disclosures
No relevant conflicts of interest to declare.