bluuebunny/crossref_metadata_2025_split huggingface dataset:
Endogenous Elevations of Short Chain Fatty Acids (SCFAs) Can Up-Regulate Embryonic Globin Gene Expression.
Authors: Bhatia Himanshu , Hallock Jennifer , Sterner Lauren , Miyazaki Toru , Dean Ann , Little Jane 
Published: April 2020

Abstract
Persistence of fetal hemoglobin can ameliorate adult beta (β)-globin gene disorders. Since SCFAs can affect embryonic and fetal globin gene expression, we examined their role during development. Murine globin gene expression, β-type (embryonic βH1, and epsilon-y, εY, and adult βmajor), and alpha (α)-type (embryonic zeta, ζ, &gt;α, adult α), were compared between wildtype (wt) and transgenic mice, in which a key enzyme for SCFA metabolism, PCCA, had been knocked out (PCCA−/−, (Miyazaki et al, 2001). E10.5 PCCA−/− yolk sac (n= 9), showed increased α, βH1 and ζ gene expression, at respectively 2-, 2.6- and 1.6-fold relative to wt (n=13, p&lt;.05), and εY gene expression, at 1.7-fold (p=0.07). The embryonic-to-adult globin gene switch was modestly delayed in yolk sacs from E12.5 PCCA−/− (n=9) vs. wt (n=4) and E 14.5 PCCA−/− (n=6) vs. wt (n=6). % embryonic β-type globin gene expression (% βH1 and εY of total β globin) was 77±6 PCCA−/− and 74±3 wt at E12.5, p=n.s., and 42±13 PCCA−/− and 21±3 wt at E14.5, p&lt;.05; % emvbryonic α-type expression (% ζ of total α) was 32±3 PCCA−/−, 25±1wt at E12.5, p&lt;.05 and 7±2 PCCA−/− and 4±1 wt at E14.5, p&lt;.05). Embryonic globin gene expression in E 12.5 and 14.5 fetal livers was not different between PCCA−/− and wt embryos. Cultures of pooled E14.5 wt fetal liver cells (FLCs, n=4 separate experiments), however, suggested that embryonic globin genes can be activated in FLCs. The percent of total β-type globin gene expression that was embryonic after culture with butyrate (1mM) was 11.6±2.6%, with propionate (2.5 mM) was 3.6±0.2%, and insulin/erythropoietin or basal media was 0.03±0.03% and 0.42±0.26% respectively (p&lt;.05 relative to SCFAs). Dose-response with propionate (n=2 seaparate experiments) suggest inadequate endogenous propionate levels for activation in PCCA −/− fetal liver, as % embryonic β-type globin gene expression rose above basal levels only at concentrations of 1 to 5 mM (2.5 mM maximal) but not at &lt;0.6 mM. We conclude that endogenous SCFAs, at levels achievable in vivo can activate embryonic globin gene expression during development in the murine yolk-sac. However, higher levels than achievable endogenously currently are necessary to produce this effect in murine fetal livers.
Regulation of TLR Expression and Activation in Human Mesenchymal Stromal Cells by IFN-a, IFN-g and TNF-a.
Authors: François Moïra , Romieu-Mourez Raphaëlle , Boivin Marie-Noëlle , Galipeau Jacques 
Published: April 2020

Abstract
Members of the toll-like receptor (TLR) family bind to pathogen-associated molecular patterns and subsequently induce the activation of the innate and adaptive immune responses. We investigated here the expression and role of TLRs in mesenchymal stromal cells (MSC), which are bone marrow-derived mesenchymal progenitors with recently described antigen presenting cell (APC) properties [Stagg et al, Blood (March 2006), Romieu et al., JI (in press)]. We observed that human MSCs express basal levels of TLR3 and TLR4, and these responded to their respective ligands, e.g. polycytidylic acid (poly I:C) or lipopolysaccharide (LPS), by the production of IL-6, IL-8 and IL-1b but not of active IL-12 p70 dimer. Exposure of MSCs to TNF-a increased the expression of TLR2 and TLR4 while IFN-a strongly upregulated TLR3 expression. By contrast, IFN-g had only a marginal effect on the expression of any TLR. IFN-a and poly I:C, as well as IFN-g and poly I:C or LPS were observed to work in synergy for the up-regulation of expression of several cytokines or inflammatory mediators, such as IL-12 p35 (IL-12A), TNF-a, RANTES, TRAIL, IFN-b and NOS2A in MSC. In contrast, none of these treatments induced the upregulation of IL-12 p40 (IL-12B), which may explain the lower levels of IL-12 p70 produced in MSCs compared to primary macrophages. Since c-Rel/p50 NF-kB dimers as well as IFN-induced IRF-1 were shown to control IL-12B and/or IL12A promoters, respectively, we next investigated expression of these transcription factors in MSCs. Results showed that MSC expressed the ubiquitous RelA NF-kB subunit but displayed significantly lower expression of c-Rel compared to macrophages. IRF-1 was upregulated following treatments with IFN-g or, to a lesser extent, with IFN-a. Therefore, the lower levels of IL-12 p70 seen in MSCs compared to macrophages may be explained by the absence of c-Rel in MSCs, which hinders the upregulation of IL-12A and IL-12B after exposure to single TLR ligands. However, the induction of IRF-1 by IFN-g or IFN-a may explain the synergistic effect observed in the upregulation of IL-12A in MSC treated with IFN-g or IFN-a and TLR ligands. In conclusion, TLR expression in human MSCs plays a role in their immune activation, although the outcome of this activation is different from the result of macrophage activation by TLR.
IREM-1 Is a Cell Surface Receptor Expressed in AML Blasts: Potential for Targeted Immunotherapy in AML.
Authors: Zhao Xiaoxian , Korver Wouter , Singh Shweta , Hsi Eric D. , Abo Arie 
Published: April 2020

Abstract
Background: Acute myeloid leukemia (AML) in adults is associated with a poor prognosis, with many patients suffering from treatment-resistant relapse. Novel targeted therapies are needed. Immune Receptor Expressed by Myeloid Cells (IREM-1), a member of the CD300L gene family, is an inhibitory receptor expressed by myeloid cells. We have generated monoclonal antibodies (mAbs) specific for IREM-1, characterized IREM-1 expression in AML blasts and explored the therapeutic potential of these antibodies in AML.
Methods: mAbs were generated against the extracellular domain of IREM-1. Fresh peripheral blood or bone marrow cells from AML patients, healthy donors, or cell lines OCI-AML-2 (IREM-1+) and K562 (IREM-1−) were used for expression studies by flow cytometry (FC) with FITC-conjugated mAbs. Immunohistochemistry (IHC) was carried out on Tissue Microarrays (TMAs) of normal human tissues. mAbs were evaluated for their ability to induce lysis in Complement-dependent cytotoxicity (CDC) assays against cell lines in vitro and AML patient cells ex vivo.
Results: By IHC, normal tissues lacked expression of IREM-1, with the exception of spleen and tonsil. However, using FC, expression of IREM-1 was demonstrated on monocytes, granulocytes and dendritic cells, but not on lymphocytes and platelets. FC of AML blasts showed that 18/24 (75%) cases expressed IREM-1 (range: 23.8–93.2%). The 6 negative cases of IREM-1 include 2 AML cases of inv (16) (p13q22). IREM-1 was not detectable in acute lymphocytic leukemia (ALL) blasts (n=5). CD34+ hematopoietic stem cells of control bone marrows were largely negative (n=7), only one case shown low level expression. Functionally, anti-IREM-1 antibodies mediated CDC in OCI-AML-2, while no effect was seen in IREM-1 negative K562 cells. Human embryonic kidney 293 cells only became susceptible to anti-IREM-1 antibody mediated killing after transfection with IREM-1. Furthermore, an anti-IREM-1 mAb showed dose-dependent CDC activity in all IREM-1+ AML cases tested (n=4), while no CDC activity was observed against IREM-1− AML cases (n=3) and ALL blasts (n=3).
Conclusion: Our study identifies IREM-1 as a novel cell surface antigen expressed in majority of AML blasts. We have generated mAbs that are capable of mediating CDC activity against AML blasts. Currently we are testing the killing activity of anti-IREM-1 chimeric Ab and undertaking mouse xenograft model to evaluate the therapeutic impact of this target in vivo, which would offer opportunities for therapeutic intervention.
A Single Nucleotide Polymorphism in CD19 Defines a Novel Target for Immunotherapy of B Cell Malignancies with CD4+ Cytotoxic T Cells.
Authors: Spaapen Robbert M. , Lokhorst Henk M. , van den Oudenalder Kelly , Otterud Brith , Dolstra Harry , Leppert Mark , Minnema Monique C. , Bloem Andries C. , Mutis Tuna 
Published: April 2020

Abstract
The Graft-versus-Tumor (GvT) effect of HLA-matched allogeneic stem cell transplantation (allo-SCT) is largely mediated by donor-derived alloreactive CD4+ and CD8+ T cells. Major targets of this curative effect are the minor Histocompatibility antigens (mHags) expressed on the malignant cells. Here we report the first mHag encoded by a hematopoietic gene and recognized by HLA class II (HLA-DQA1*05/B1*02)-restricted CD4+ T cells. This antigen is encoded by a single nucleotide polymorphism (SNP) in the B cell lineage-specific CD19 gene, a highly important target antigen for immunotherapy of almost all B cell malignancies. We identified this antigen using a novel and powerful genetic strategy, in which a phenotype-genotype correlation scanning was the key step for fine-mapping the genetic locus defined by pair-wise linkage analysis. In functional assays, CD4+ T cells specific for the CD19L-encoded mHag mediated effective peptide-dependent maturation of DCs and polarized them to produce significant levels of interleukin-12. In another assay, the CD19L-mHag-specific T cells facilitated the proliferation of a CD8+ mHag-specific T cell clone in an antigen-dependent manner. Even more important, they also lysed CD19L-positive malignant cells, illustrating the therapeutic advantages of targeting this CD19 L -derived, HLA class II-restricted mHag. The currently available immunotherapy strategies enable the exploitation of these therapeutic effects within and beyond allo-SCT settings.
Leukemia-Reactive Cytotoxic CD8+ and CD4+ T-Cells Specific for Novel WT-1 Epitopes Are Generated In Vitro by Sensitization with Overlapping Pentadecapeptides (15-mers) Spanning the Wilms Tumor Protein.
Authors: Doubrovina Ekaterina , Dupont Jacob , Kanaeva Elena , O’Reilly Richard J. 
Published: April 2020

Abstract
The Wilms Tumor protein, WT-1 is differentially expressed in 60–80% of acute leukemias and CML. Evidence also suggests WT-1 expression in clonogenic leukemia stem cells, making WT-1 an attractive target for T-cell therapy. However, to date, only a limited number of immunogenic epitopes have been identified. To address this limitation, we employed a pool of 15-mers with 10 amino acid overlaps spanning the complete sequence of WT-1 loaded on autologous monocyte derived dendritic cells or EBV transformed BLCL to sensitize T-cells from normal donors of varied HLA genotypes. Specific 15-mers eliciting WT-1 specific responses were identified by quantitating IFNγ+ CD8+ or CD4+ T-cell responses to secondary stimulation with 15-mer subpools organized in a mapping grid with single overlaps. Phenotypes of T-cells producing IFNγ+ in response to identified specific 15-mers were determined by FACS analysis. We identified the HLA class I or II allele presenting each epitope by assessing the cytolytic activity and/or cytokine production of the pooled-peptide sensitized T-cells in response to 2° stimulation with a panel of PHA blasts loaded with the identified 15-mer, each sharing a single HLA allele with the T-cell donor. T-cells from each of 15 donors tested generated WT-1 peptide reactive T-cells in response to the WT-1 peptide pool. Of 23 epitopes identified that elicited T-cell responses, 19 were heretofore unrecognized epitopes presented by HLA A0201(N=4), A0301(N=2), A2402 (N=2), B3501(N=3), B0701(N=1), B4402(N=2), B3808(N=1), DRB, 0401(N=2), DRB10402(N=1) and DRB1 0701(N=1). Each of the 15 new epitopes presented by class I alleles elicited IFNγ+ CD8+ T-cells that were also cytotoxic against leukemic blasts sharing the restricting allele. All four epitopes presented by class II HLA alleles elicited IFNγ+ CD4+ T-cell responses, of which 2 were also cytotoxic against HLADR+ leukemia blasts expressing the restricting allele. Responses to these newly identified epitopes were comparable to or exceeded those elicited by the known immunogenic peptide epitope 126–134RMFPNAPYL presented by HLAA0201. Like RMF-specific T-cells, T-cells specific for 4 of the new epitopes tested to date do not exhibit activity against normal hematopoietic progenitor cells. Thus, by sensitizing T-cells from unselected normal donors, with autologous DCs loaded with a pool of overlapping 15-mers spanning WT-1, we have generated T-cells specific for a series of new epitopes of WT-1, each, presented by a distinct class I or II HLA allele, which also exhibit HLA-restricted cytotoxic against WT-1+ leukemic cells. Clinical trials of adoptive therapy with WT-1 specific T-cells generated by this approach are being initiated for adoptive therapy.
A Novel Measurement of the Rate of Spontaneous Mutations in Human Myeloid Cells.
Authors: Araten David J. , Mulloy James C. , Krejci Ondrej , DiTata Kimberly 
Published: April 2020

Abstract
The mutation rate (μ) is likely to be a critical parameter in leukemogenesis, but has not previously been measured in human myeloid cells. Using the PIG-A gene as a sentinel, we have recently measured the mutation rate in B-lymphoblastoid cell lines (BLCLs) and shown that it is elevated in cell lines derived from lymphoid malignancies. Here we have measured μ in normal human CD34+ cells derived from cord blood samples that have been transduced with an AML-ETO fusion gene, growing under the influence of cytokines. PIG-A is an X-linked gene essential for the synthesis of glycosylphosphatidylinositol (GPI). Due to X-inactivation in females and hemizygosity in males, a single mutation can produce the GPI (−) phenotype. From patients with PNH, it is known that a broad spectrum of mutations can inactivate PIG-A. Mutants do not express GPI-linked surface proteins such as CD55 or CD59 and do not bind to the FLAER reagent (which binds GPI directly), but do express transmembrane proteins (e.g. CD45). To measure the rate of new mutations in vitro, we first flow-sorted each culture to collect CD59(+) cells, in order to exclude pre-existing mutants. Then we expanded the collected cells over 3 weeks; using cell counts, we calculated population doublings in culture (d), which ranged from 5.1 to 7.4. Cells were then stained sequentially with biotinylated FLAER, murine anti-CD55 and anti-CD59 antibodies, rabbit anti-mouse PE and streptavidin-PE secondary conjugates, and CD45-FITC. Live cells were identified by forward and side scatter, propidium iodide exclusion, and expression of CD45. The mutant frequency (f) was calculated as the number of GPI (−) cells [which do not bind anti-CD55, CD59 or FLAER] divided by the total number of cells analyzed. An average of 1.2 million cells were analyzed to detect rare mutants. The mutation rate was calculated by the formula μ = f/d. Among 5 cultures derived from 2 different cord blood samples, the mean mutation rate was 12.4 × 10−7 (range 5.1 to 21.4 × 10−7) per cell division. This value is consistent with mathematical models of the mutation rates in human populations as well as our previous experimental determination of μ in BLCLs. We also analyzed 6 cell lines in which p53 had been disrupted by various methods, including shRNA, a dominant-negative p53 mutant, and introduction of HPV E6/E7. Overall, disruption of p53 resulted in a 30% increase in the mutation rate (p=.02). We conclude that spontaneous mutations in human myeloid cultures are rare but measurable and that disruption of p53 results in a modest increase in μ. We predict that the ability to measure this critical parameter in human hematopoietic cells will facilitate the investigation of the role of specific oncogene and tumor suppressor mutations in inducing hypermutability.
A New Fusion Gene NUP98-IQCG Identified in an Acute T/Myeloid Leukemia with t(3;11)(q29q13;p15) Translocation.
Authors: Pan Qin , Zhu Yong-Jin , Gu Bai-Wei , Cai Xun , Bai Xue-Tao , Yuan Hai-Yang , Zhu Jiang , Chen Zhu , Xue Yong-Quan , Chen Sai-Juan 
Published: April 2020

Abstract
NUP98 has been involved in multiple recurrent chromosome rearrangements in leukemia. We identified a novel fusion between NUP98 and IQ motif containing G (IQCG) from a de novo acute T/myeloid leukemia harboring t(3;11)(q29q13;p15). IQCG has 2 putative coiled-coil domains and an IQ domain. The FG repeat from NUP98 and the coiled-coil domain from IQCG were retained in the fusion protein. We demonstrated that NUP98-IQCG could form homodimer or heterodimerize with either NUP98 or IQCG, bind Co-activators and/or Co-repressors, and show transcriptional activity in vitro. Expression of NUP98-IQCG inhibited 32Dcl3 cell apoptosis induced by Ara-C, and partially blocked granulocyte differentiation induced by G-CSF. Colony forming assay also indicated that NUP98-IQCG was able to stimulate proliferation and partially block differentiation of hematopoietic stem/progenitor cells. We conclude that NUP98-IQCG may play an essential role in leukemogenesis by stimulating progenitor cell proliferation and inhibiting differentiation and apoptosis.
Intensively Timed Induction (ITI) Chemotherapy in Adults with Acute Myelogenous Leukemia (AML).
Authors: Rytting Michael , Verstovsek Srdan , Garcia-Manero Guillermo , Ravandi-Kashani Farhad , Pierce Sherry , Estey Elihu 
Published: April 2020

Abstract
The Children’s Cancer Group (CCG) demonstrated that children with AML have longer event free survival (EFS) and disease free survival (DFS), 42 +/− 7% and 55 +/− 9%, at a minimum of two years off of therapy when randomly assigned to ITI rather than standard chemotherapy (Woods, et al Blood 1996 Jun 15;87(12):4979–89). This led us to administer an almost identical regimen of ITI chemotherapy to 50 adults less than 50 years of age with untreated AML, excluding patients (pts) with inv16 or t(8;21). Each pt who remains alive in 1st complete remission (CR) has been followed for more than 3 years: the risk of relapse decreases sharply after this time (de Lima, et al. Blood 1997 Dec 15;90(12):4719–24). Daunorubicin 20 mg/m2/day intravenously (IV) on days 1–4 and 11–14, cytarabine 200 mg/m2/day IV on days 1–4 and 11–14, etoposide 100 mg/m2/day IV on days 1–4 and 11–14 and dexamethasone 6 mg/m2/day PO days 1–4 and 11–14 were given initially along with intrathecal cytarabine 70 mg on day 1 and 11 (DCTER regimen). G-CSF started on day 7 and continued until the ANC was more than 1000 for 2 days. The day 11 chemotherapy was given unless there was hypotension or clinical colitis. All pts started therapy in a protected environment. Pts in CR after the above chemotherapy repeated the induction course. After recovery, cytarabine 3 g/m2 every 12 hours for four doses was infused on days 0–3 and on days 12–15. L-asparaginase 6000 IU/m2 IV was given at hour 18 on days 3 and 15. The cytarabine and L-asparaginase were repeated upon recovery of the granulocyte count. The study was ongoing from 2/15/02 to 8/10/04. The 50 pts, median age 41 years, represented 62% of all pts eligible during this time period. Table 1 compares results in the 50 DCTER pts with the results in 35 non-core binding factor AML pts, median age 41 years, given idarubicin/cytarabine (IA) for induction and consolidation without intensive timing. The results in the IA pts treated from 1991–1993 were similar to those treated 1994–2001 (proportion alive in CR 16% and 19%). The lower CR rate in pts given DCTER reflected an increased death rate in the first 12 weeks after starting treatment. This was counterbalanced by a higher rate of potential cure, considering all 85 pts (95% confidence interval for the true difference [−.09, .34] suggesting that a difference of 26% is as plausible as no difference since .26 and 0 are each equidistant from the midpoint of the confidence interval). A larger cohort is needed to know if the higher early risk associated with DCTER is potentially justified by a better long-term prognosis.
Table 1 Patients Performance Status &gt;1 Unfavorable Cytogenetics CR Proportion alive in CR at 3 years DCTER 50 14% 24% 69% 30% IA 35 0% 23% 80% 17%
Generic and Branded Low Molecular Weight Heparins Can Be Differentiated Based on Biologic and Pharmacologic Assays.
Authors: Jeske Walter P. , Ackerman Paul , Drenth Amanda , Walenga Jeanine M. , Bakhos Mamdouh 
Published: April 2020

Abstract
Several generic versions of the low molecular weight heparins enoxaparin and dalteparin have become available in different parts of the world and several products are currently under review by the FDA. At this time, there are no regulatory guidelines for defining equivalence of complex biologic drugs. Because of their complexity and hybrid nature (biologic and chemical), the available generic versions may not be acceptable by using currently applicable generic criteria. Due to their complex nature, the pharmacologic effect of these may not be similar to the branded product. Therefore, additional methods are needed to equate these drugs with the branded products. In this study, multiple lots of Lovenox or generic enoxaparin such as Lupenox (Lupin Pharma, India), Dripanina (Ariston, Brazil), Dilutol (Lazar, Argentina), Clenox (Pharmayect, Columbia), and Cutenox (Gland Pharma, India) and generic versions of dalteparin such as Daltehep (Gland Pharma, India) were supplemented to human plasma in the absence or presence of protamine sulfate (PS). Anticoagulant activity was measured using the aPTT, Heptest, PiCT assays. Antithrombin and anti-factor Xa activity were assessed by amidolytic assays. Effects on thrombin generation and the activation of thrombin activatable fibrinolytic inhibitor (TAFI) were determined by functional assay. At prophylactic levels of LMWH, no differences in anticoagulant or antiprotease activities were observed between generic and branded LMWHs. At therapeutic concentrations, significantly higher anticoagulant and antiprotease activities were observed with Lovenox. Assay-dependent variations in PS neutralization of antiprotease and anticoagulant activities were observed at higher concentrations of the generic versions of enoxaparin and dalteparin. Significant differences were noted in the effects of these agents on thrombin generation and the activation of TAFI. Current generic LMWHs possess some differences from branded LMWHs in known biologic properties of heparin. Variations in PS neutralization raise the question as to whether the interaction with other plasmatic proteins also differs from that of the branded drug. These results emphasize a need to consider multiple functional parameters when defining bioequivalence of complex biologic drugs and underscore the importance of further pharmacologic studies involving animal models and human clinical trials. Such approaches can be incorporated in the recommendations to develop guidelines for the approval of the generic versions of low molecular weight heparins.