bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bendamustine-Bortezomib-Desametasone (BVD) in the Management of Relapsed and Refractory Multiple Myeloma : A REAL-Life Experience
Authors: Cerchione Claudio , Catalano Lucio , Pareto Anna Emanuele , Basile Santina , Marano Luana , Peluso Ilaria , Simeone Luigia , Vitagliano Orsola , Nappi Davide , Palmieri Salvatore , Rocco Stefano , Ferrara Felicetto , Pane Fabrizio 
Published: October 2019

Abstract
Introduction :
Bendamustine is a bifunctional alkylating agent, with low toxicity, proved to be effective in relapsed, refractory and in new diagnosed Multiple Myeloma (MM).
It has been evaluated efficacy and tolerance of Bendamustine, in combination with bortezomib-dexamethasone (BVD) in patients with relapsed and refractory MM (rrMM), whose prognosis is particularly severe. A regional retrospective real-life analysis of patients with rrMM who had been treated with BVD as salvage therapy has been performed.
Methods :
56 patients (31 M/25 F), with rrMM, median age at diagnosis 57.3 years (r. 36-82), median age at start of treatment 61.8 years (r.37-83) treated with several lines of treatments (median 6, r. 2-11), every refractory to all the drugs previously received (also Bortezomib), received BVD (Bendamustine 90 mg/sqm days 1,2; Bortezomib 1.3 mg/sqm days 1,4,8,11, Dexamethasone 20 mg days 1,2,4,5,8,9,11,12, Pegfilgrastim day +4) every 28 days, until progression.
ISS was equally distributed, and cytogenetic was evaluable in 12 patients, and in particular one del13q and one t(11;14). All the patients had previously been treated with schedule containing bortezomib and IMIDs, and 30% had also received radiotherapy. 67% of them had undergone at least to a single auSCT. All patients were relapsed and refractory to last therapies received before BVD.
Results :
Bendamustine was well tolerated, with grade 3 transfusion-dependent anemia in 41% of patients, and 37% grade 3 neutropenia (no ospedalization was required, no septic shocks were observed). No severe extrahematologic toxicity was observed, only grade 1 gastrointestinal side effect (nausea), treated by common antiemetic drugs.
According to IMWG, after a median follow-up of 14 months (r.2-36), ORR was 64% (36/56: 4 CR, 7 VGPR, 16 PR, 9 MR) with 8 PD and 12 patients in SD, which can be considered as an impressive result in this subset of rrMM patients. In particular, for 11 patients, BVD was, after having achieved at least a PR, a bridge to second auSCT, and for two patients a bridge to alloSCT.
Median time to response was 1.2 months (range, 1-3), median OS from diagnosis was 62.7 months (range, 6-151), median OS from start of Bendamustine was 9.8 months (range 2-36).
Conclusion :
BVD has shown significant efficacy in a particularly severe setting of patients, relapsed and refractory to all available therapeutic resources, and, in particular cases, it could be considered as a bridge to a second autologous or allogenic SCT.

Disclosures
No relevant conflicts of interest to declare.
Allogeneic Hematopoietic Cell Transplantation for Lymphoma: Baseline and Post-Transplant Prognostic Factors
Authors: Ko Sun-Hye , Lee Jung-Hee , Lee Je-Hwan , Park Han-Seung , Choi Eun-Ji , Lee Young-Shin , Kang Young-Ah , Lee Kyoo-Hyung 
Published: October 2019

Abstract
Background: Allogeneic hematopoietic cell transplantation (HCT) has been increasingly used for the treatment of lymphoma. The outcomes of HCT in lymphoma depend on baseline patient characteristics including chemosensitivity and International Prognostic Index (IPI). Several studies have shown that post-transplant occurrence of acute or chronic graft-versus-host disease (GVHD) and immune recovery might be associated with the outcomes. In this retrospective study, we investigated baseline and post-transplant prognostic factors in lymphoma patients receiving allogeneic HCT.
Patients and methods: Between May 1998 and December 2015, a total of 61 patients underwent allogeneic HCT for lymphoma and the median age was 39 years (range, 16-62 years). Thirty four patients (55.7%) had chemo-sensitive disease and 24 patients had received autologous HCT. Fifty-six of 61 patients received reduced-intensity conditioning regimens. We evaluated tumor response, overall survival (OS), progression-free survival (PFS), non-relapse mortality (NRM), and event-free survival (EFS) after allogeneic HCT along with potential prognostic factors including GVHD and immune reconstitution.
Results: Objective tumor response after HCT was observed in 41 (67.2%; complete 30 and partial 11). The 5-year probabilities of OS, NRM, PFS, and EFS were 30.7%, 23.5%, 41.3%, and 24.0%, respectively. Among the baseline characteristics, chemosensitivity had a significant impact on OS, NRM, and EFS. Persistent disease status at allogeneic HCT had an adverse effect on OS and low IPI risk category at HCT was associated with longer OS and EFS. HCT co-morbidity index ≥1 was an independent prognostic factor for higher NRM. Grade III-IV acute GVHD was associated with lower OS and higher NRM. Severe chronic GVHD had higher OS (60.0% vs. 19.8%, P=0.002), PFS (68.8% vs.35.9%, P <0.001), and EFS (46.7% vs. 19.1%, P<0.001) than those without severe chronic GVHD. Patients who achieved higher CD3+, CD4+ or CD8+ cell counts on post-transplant 1 month showed significantly higher OS and EFS than those who did not.
Conclusion: Chemosensitivity was the most important prognostic factor in allogeneic HCT for lymphoma. Acute GVHD had unfavorable impact, whereas, chronic GVHD had favorable impact on post allogeneic HCT outcomes. Early immune recovery could predict allogeneic HCT outcomes.

Disclosures
No relevant conflicts of interest to declare.
Opposing Effects of PD-1/PD-L1/L2 Engagement and IFN-γ/TNF-α in the Treatment of AML w/ Anti-CD33 Chimeric Antigen Receptor-Modified T Cells
Authors: Basham Jacob Halum , Geiger Terrence L. 
Published: October 2019

Abstract
Chimeric antigen receptor-modified T lymphocytes (CART cells) have shown benefit as an adjuvant immunotherapy in the treatment of B cell malignancies. This success of re-targeted T cells has not been extended to other hematologic malignancies. We have developed an immunotherapeutic approach to treat acute myeloid leukemia (AML) using CAR T cells re-directed against the myeloid-specific antigen CD33 (CART-33). CART-33 cells are potent and specific in eliminating AML cells in vitro and in vivo. Despite this, CART-33 cells have shown poor in vivo expansion and persistence in NOD-SCID IL2rγ (-/-) (NSG) AML xenograft models. To address the reason for this, we assessed the impact of AML-expressed programmed death ligands 1 & 2 (PD-L1/2) on CART-33 cell activity. PD-L1 inhibits T cell functions upon binding PD-1, which is upregulated with T cell activation. Less is known about PD-L2's effect. Interferon-gamma (IFN-γ), a primary effector cytokine secreted by CD4+ and CD8+ effector T cells, is a known potent inducer of PD-L1 on AML blasts. Using AML cell lines U937, Oci-AML3, CMK, and MV4-11 we show that IFN-γ, TNF-α, and activated CART-33 supernatant can induce up-regulation of PD-L1 and PD-L2 on AML. IFN-γ and TNF-α synergize strongly in up-regulating PD-1 ligands on AML. The kinetics and induction of PD-L2 are distinct from that of PD-L1. Although PD-L1 is well documented to suppress T cell function via ligation of T cell expressed PD-1, induction of PD-L1/L2 had no effect on the cytolytic activity of CART-33 cells against AML in short term (<48 h) cultures. Paradoxically, 24 hr pre-treatment of AML with either IFN-γ or CART-33 supernatant increased AML susceptibility to killing by CART-33 cells despite elevated expression of PD-L1/L2 by AML. Our results highlight the regulatory complexity of AML cytolysis by re-targeted T lymphocytes, and argue that tumor-expressed PD-L1 and PD-L2 impacts the sustainability, but not short-term killing activity, of adoptively transferred CAR T cells in the treatment of AML.

Disclosures
No relevant conflicts of interest to declare.
The Development of a qPCR Assay for the Evaluation of the Dendritic Cell Chimeric Antigen Receptor Transcriptome
Authors: Leyfman Yan , Daniyan Anthony , Brentjens Renier J. 
Published: October 2019

Abstract
Chronic lymphocytic leukemia (CLL) is a hematological malignancy of B cells that accounts for a third of all new leukemia cases, but is incurable with standard first-line chemotherapies. Hematopoietic stem cell transplants offer the only curative treatment but are restricted to those with a matching donor. The recent advent of adoptive transfer therapy utilizing Chimeric Antigen Receptor (CAR) T cells, which are synthetic receptors that combine the target-specificity of antibodies to the effector capabilities of T cells, have emerged as a promising anticancer therapy due to its targeted tumoricidal activity. Clinical trials utilizing CD19-targeted CAR T cells have demonstrated strong efficacy against acute B-cell leukemia, but those performed in CLL have not been as successful for unclear reasons. This may be due to the dysfunctional tumor microenvironment which renders key immune effectors ineffective. Dendritic cells (DCs), a potent antigen presenting cell that functions as a bridge between the innate and adaptive immune systems, are rendered functionally and quantitatively deranged with an impaired ability to present antigen and thereby stimulate immunogenic responses. These limitations can potentially be rectified through an effective recruitment of an endogenous antitumor immune response and thus we hypothesize that genetically modified DCs should be capable of repolarizing the tolerogenic microenvironment induced by CLL toward one of immunogenicity to promote tumor eradication. In this study, we developed a qPCR assay to assess the DC CAR transcriptome. Because our CD19-targeted DC CARs with a CD40 intracellular domain signal via NF-κB to activate downstream elements, we needed to first assess the fidelity of our assay. Using the immortalized mouse DC cell line JAWS II, which lacks CD40, we used a potent TLR4 agonist, MPL, to stimulate our DCs through MyD88, an alternative pathway that also activates NF-κB. We saw elevated expression of downstream NF-κB elements, including IL-6, IL-10, IRF3, and IRF7. Establishing the sensitivity of our assay, we next evaluated the transcriptome of DC CARs by coculturing our CD19-targeted DC CARs containing a CD40 intracellular domain with HEK 293 cells, a human embryonic kidney cell line, retrovirally transduced with a human CD19 antigen to activate our CAR DCs and assess downstream gene expression. We observed expected elevation of downstream NF-κB elements including IL-6, IL-10, NOS2, IRF3, IFN-γ, TNFα, IL12p40, and TGF-β, indicating the viability of this assay. These positive results confirm the multidisciplinary potential of our assay as a promising technology that will allow us to better augment future CARs.

Disclosures
Brentjens: Juno Therapeutics: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding.
Regulation of T-Cell Activation and Differentiation
Authors: Vahedi Golnaz 
Published: October 2019

Abstract
Enhancers regulate spatiotemporal gene expression and impart cell-specific transcriptional outputs that drive cell identity. Super-enhancers (SEs), also known as stretch-enhancers, are a subset of enhancers especially important for genes associated with cell identity and genetic risk of disease. CD4+ T cells are critical for host defence and autoimmunity. Here we analysed maps of mouse T-cell SEs as a non-biased means of identifying key regulatory nodes involved in cell specification. We found that cytokines and cytokine receptors were the dominant class of genes exhibiting SE architecture in T cells. Nonetheless, the locus encoding Bach2, a key negative regulator of effector differentiation, emerged as the most prominent T-cell SE, revealing a network in which SE-associated genes critical for T-cell biology are repressed by BACH2. Disease-associated single-nucleotide polymorphisms for immune-mediated disorders, including rheumatoid arthritis, were highly enriched for T-cell SEs versus typical enhancers or SEs in other cell lineages. Intriguingly, treatment of T cells with the Janus kinase (JAK) inhibitor tofacitinib disproportionately altered the expression of rheumatoid arthritis risk genes with SE structures. Together, these results indicate that genes with SE architecture in T cells encompass a variety of cytokines and cytokine receptors but are controlled by a 'guardian' transcription factor, itself endowed with an SE. Thus, enumeration of SEs allows the unbiased determination of key regulatory nodes in T cells, which are preferentially modulated by pharmacological intervention.

Disclosures
No relevant conflicts of interest to declare.
Surviving Change: The Metabolic Journey of Hematopoietic Stem Cells
Authors: Passegue Emmanuelle 
Published: October 2019

Hematopoietic stem cell (HSC) activation is accompanied by mitochondria activation and a shift in metabolic activity from glycolysis to OXPHOS, which provides energy and increases the production of ROS and other mitochondrial metabolites that can act both as signaling molecules and substrates/co-activators for epigenetic enzymes. Metabolically activated HSCs are poised to undergo lineage priming and produce different lineage-biased multipotent progenitors (MPP). However, activated HSCs must also return to quiescence to maintain the stem cell pool. In this context, autophagy plays an essential role by clearing activated mitochondria to allow OXPHOS-driven HSCs to efficiently revert to a mostly glycolysis-based quiescence metabolism. Without autophagy, HSCs display an overactive OXPHOS-driven metabolism that promotes myeloid-biased differentiation and loss of stemness, likely as a direct consequence of epigenetic reprogramming. At steady state, blood production reflects the differential generation by HSCs of a small number of myeloid-biased MPP2/3 and a large number of lymphoid-biased MPP4, which both contribute to myeloid output. In contrast, during blood regeneration, activated HSCs are induced to overproduce MPP2/3, and MPP4 are reprogrammed towards almost exclusive myeloid output in large part due to cytokine stimulations and the triggering of specific regulatory pathways. Altogether, the metabolic activation of HSCs and the remodeling of the MPP compartment represent emergency myelopoiesis pathways that are transiently activated during regeneration, and are continuously triggered in myeloid disease conditions.

Disclosures
No relevant conflicts of interest to declare.
Specialized Ribosomes: A New Frontier in Gene Regulation, Organismal Biology, & Evolution
Authors: Barna Maria 
Published: October 2019

Abstract
The central dogma of molecular biology has for decades served as an explanation for the flow of genetic information within a biological system. In so far as the normal flow of biological information from mRNA to protein, the ribosome has been perceived to decode the genome with essentially machine-like precision; serving as an integral but largely passive participant in the synthesis of all effector proteins across all kingdoms of life. Importantly, a large class of human diseases collectively known as 'ribosomopathies' are characterized by mutations in ribosome components that lead to devastating human conditions including bone marrow failure for which the underlying molecular basis remains poorly understood. In this respect, our research has changed the view that ribosomes carry out largely rote-like functions by demonstrating that not all of the millions of ribosomes within each cell are the same and that ribosome heterogeneity provides a novel means for diversity of the proteins that can be produced in specific cells, tissues, and organisms. I will present our work centered on providing a roadmap for the characterization of ribosome composition at a single cell level and during cellular differentiation. We employed a highly quantitative mass spectrometry-based approach to precisely quantify the abundance of each ribosomal protein (RP) as well as a large cohort of auxiliary ribosome associating factors belonging to actively translating ribosomes within embryonic stem cells. This led to the identification of a subset of ribosomes that are heterogeneous for RP composition. To further address the functional role of ribosome heterogeneity in translational control of the mammalian genome, we employed CRISPR/Cas9 to endogenously tag and purify heterogeneous ribosome populations. We then developed an adapted ribosome profiling method to precisely quantify and characterize the nature of mRNAs translated by distinct heterogenous ribosomes genome-wide. This led to the identification of subpools of transcripts, critical for key cellular processes including cell signaling, metabolism, growth, proliferation and survival, which are selectively translated by specific types of ribosomes. Most remarkably, there are specific metabolic pathways where almost every single component is selectively translated by specialized ribosomes demarcated by a single RP. I will further present recent findings on the mechanisms by which ribosome-mediated control of gene expression is encoded by structured RNA regulons within 5'UTRs. Together, these studies reveal a critical link between ribosome heterogeneity and specialized translational control of the mammalian genome, which adds an important layer of control to the post-transcriptional circuitry of gene regulation and may be critically perturbed in human diseases.

Disclosures
No relevant conflicts of interest to declare.
Nucleic Acids and Protein Metabolism in Acute Leukemia Cells
Authors: GAVOSTO FELICE , MARAINI GIOVANNI , PILERI ALESSANDRO 
Published: October 2019

Abstract
DNA, RNA and protein metabolism was investigated by means of a high resolution autoradiographic technic in normal and acute leukemia blast cells by studying the incorporation of tritiated thymidine, uridine, leucine and phenylalanine. A strikingly lower percentage of cells labelled with thymidine was demonstrated in acute leukemia and was interpreted as evidence of a decreased proliferative capacity. A very significantly lower uptake of uridine, leucine and phenylalanine was detected in acute leukemia cells.
In normal and leukemic cells, amino acid incorporation occurred both in the nucleus and in the cytoplasm; uridine was incorporated exclusively in the nucleus during the first hour of incubation and the cytoplasm became labelled only in a later period.
The constant ratio between uridine and amino acid incorporation detected in normal myeloblasts was always altered in acute leukemia cells.
The lower RNA and protein metabolism and its dissociation in acute leukemia cells was discussed as related to the well-known maturation defect of these cells.
Studies on Agranulocytosis. III. The Reduced Glutathione (GSH) Content of Leukocytes of Normals and Patients Recovered from Agranulocytosis
Authors: PISCIOTTA ANTHONY V. , DALY MARY 
Published: October 2019

Abstract
1. A method for the direct determination of GSH in leukocytes is described. Treatment with alkali (0.5 M. NaOH) effects complete solution of the white cells and after deproteinization (5 per cent HPO3), the GSH is determined by the sensitive "alloxan 305" procedure. Control studies showed that under the conditions of the test, GSH is not affected by the alkali and no splitting of soluble minus SH from protein occurs.
2. Using this method, the amount of reduced glutathione content of normal leukocytes was found to be 5.2 ± 1 mg, per 1010 WBC.
3. No differences from normal levels were detected for the leukocytic GSH of patients with mental disease and those susceptible to agranulocytosis. Incubation of whole blood with the drug which caused agranulocytosis had no effect upon the GSH content of leukocytes.
Serum Protein Binding of Myoglobin, Hemoglobin and Hematin
Authors: WHEBY MUNSEY S. , BARRETT O'NEILL , CROSBY WILLIAM H. 
Published: October 2019

Abstract
Using paper and starch-gel electrophoresis we have demonstrated binding of hemoglobin, myoglobin and hematin by human serum protein which is not a haptoglobin. This protein was present in all sera tested except that from two patients with hemolytic anemia and was also present in serum genetically deficient in haptoglobin.
The binding capacity of this protein is low and contributes only slightly to the total hemoglobin binding capacity of normal serum. However, it may represent most or all of the binding in sera from patients with hemolytic disease.
Au Unusual Lymphomatous Disease Associated with Intracytoplasmic Crystals in Lymphoplasmocytoid Cells
Authors: GOLDBERG ARTHUR F. 
Published: October 2019

Abstract
A case of an unusual lymphomatous disease associated with lymphoplasmocytoid cells containing intracytoplasmic crystals is presented. These crystals are considered to be the secretory product of these cells. They are discussed in the light of recent studies on the secretory activity of plasmocytes and related cells.
A specific diagnosis of the patient's disease is not made although the main cell type present closely resembles that occurring in Waldenström’s macroglobulinemia.
THE ANEMIA OF INFECTION, VI. THE INFLUENCE OF COBALT ON THE ANEMIA ASSOCIATED WITH INFLAMMATION
Authors: WINTROBE M. M. , GRINSTEIN M. , DUBASH J. J. , HUMPHREYS S. R. , ASHENBRUCKER HELEN , WORTH WANDA 
Published: October 2019

Abstract
The influence of cobalt on the anemia associated with inflammation has been studied in three experiments involving observations in 108 rats.
It was found that by the simultaneous administration of cobalt the anemia associated with inflammation, as produced by the injection of turpentine, could be prevented from developing and polycythemia appeared instead.
This effect was accompanied by hypoferremia and an increase in erythrocyte protoporphyrin values similar to those encountered when anemia develops in association with inflammation.
Similar, though less marked, chemical changes were observed when only cobalt was given and polycythemia was produced.
A decrease in plasma albumin was noted in rats injected with cobalt or turpentine, or both, but this was not accompanied by an increased excretion of urinary nitrogen as measured by the urine urea and ammonia.
The observations cited are consistent with the hypothesis that cobalt favorably influences the utilization of iron for the synthesis of hemoglobin.
THE INFLUENCE OF ANEMIA ON PHAGOCYTIC FUNCTIONS AND RESISTANCE TO INFECTION IN MICE
Authors: BERRY L. JOE , HALLER EVELYN C. 
Published: October 2019

Abstract
The increase in phagocytic function of neutrophiles of mice was measured during the progressive development of anemia due to blood loss. The animals were bled from the tail, 2 per cent of total body weight being removed twice weekly for nine bleedings. Phagocytosis was 161 per cent of normal at the end of this period, and the erythrocyte totals and hemoglobin values were approximately two thirds of normal. The anemic mice were infected intraperitoneally with a suspension of Salmonella typhimurium and their survival rate was compared with normal mice similarly infected. Twenty-eight of 63 anemic mice survived and 14 of 63 normal mice survived.
In a second experiment, mice were subjected to bleeding every third day for nine bleedings, and were then infected along with a control group. One half of the anemic animals were bled for three additional bleedings after the infection. Two of 48 control animals survived, 5 of 47 anemic mice, additionally bled, survived, and 19 of 47 anemic mice, not bled after infection, survived. This suggests that the increase in phagocytosis that accompanies anemia may be, at least in part, responsible for the increased resistance to infection.