bluuebunny/crossref_metadata_2025_split huggingface dataset:
Gene Expression Profiles with Signatures of Tumor Biology and Chemotherapy Sensitivity May Provide a Novel Approach to Maximize Response to Induction Therapy in Patients with Acute Myeloid Leukemia.
Authors: Rao Arati V , Tuchman Sascha A , Potti Anil , Rizzieri David A. , Mano Hiroyuki , Delwel Ruud , Valk Peter , Lowenberg B. 
Published: April 2020

Abstract
Gene expression profiling in acute myeloid leukemia (AML) patients has previously identified prognostically important clusters driven by the presence of chromosomal lesions, genetic mutations (CEBPA), or abnormal oncogene expression (EVI1). Several of these patients despite response to induction chemotherapy have poor disease-free survival (DFS), and relapse rates, thus emphasizing the need for novel therapeutic strategies. Gene expression analysis was conducted by applying signature profiles reflecting deregulation of oncogenic signaling pathways ( Myc, Ras, Src, PI3K), profiles representing an altered tumor environment (angiogenesis, TNF-α), and signatures of chemotherapy sensitivity (adriamycin and etoposide) to a cohort of 345 newly diagnosed AML patients previously treated on protocols of the Dutch–Belgian Hematology–Oncology Cooperative group and the Japan Adult Leukemia Study Group. Four distinct clusters were identified based on oncogenic signaling and tumor environment pathways. There was a statistically significant difference in overall survival (OS) (p &lt; 0.0195) and DFS (p&lt;0.0304) between the four groups. Patients in cluster 1 had the worst OS at 13.5 months and DFS of just 8.4 months. Patients in this cluster had increased activation of Myc and PI3K pathways and low levels of Src activation. Patients in clusters 2 and 4 had the best OS and DFS. Fifty-one percent of patients in cluster 4 had a nucleophosmin mutation (NPM1 +) and demonstrated higher activation of the Ras and TNF pathways compared to cluster 2, while patients in cluster 2 had increased activation of Myc when compared to Cluster 4. Interestingly, patients in Cluster 3 were most sensitive to the anthracycline, adriamycin, yet had a fairly poor DFS of just 14.4 months. While many biologic factors may play a role, one reason for poor DFS might be increased Src activation in this cluster and also much lower rates of NPM1 (12%) mutations. We could hypothesize that patients in this cluster might benefit from induction with chemotherapy in combination with a Src-inhibitor. This analysis has demonstrated distinct patterns of altered tumor biology and chemotherapy sensitivity amongst patients with clinically relevant subtypes of AML. Additional results evaluating chemosensitivity to cytarabine, and other nucleoside analogues are pending.
Figure Figure
The Effects of Continued Azacitidine (AZA) Treatment Cycles on Response in Higher-Risk Patients (Pts) with Myelodysplastic Syndromes (MDS)
Authors: Silverman Lewis R. , Fenaux Pierre , Mufti Ghulam J , Santini Valeria , Hellström-Lindberg Eva , Gattermann Norbert , Sanz Guillermo , List Alan F. , Gore Steven D. , Seymour John F , Backstrom Jay , McKenzie David , Beach C. L. 
Published: April 2020

Abstract
Background. The international, phase III, multicenter AZA-001 trial demonstrated AZA is the first treatment to significantly extend overall survival (OS) in higher-risk MDS pts (Fenaux, Blood2007;110:817). The current treatment paradigm, which is based on a relationship between complete remission (CR) and survival, is increasingly being questioned (Cheson, Blood2006;108:419). Results of AZA-001 show CR is sufficient but not necessary to prolong OS (List, J Clin Oncol26:2008;abstr 7006). Indeed, the AZA CR rate in AZA-001 was modest (17%), while partial remission (PR, 12%) and hematologic improvement (HI, 49%) were also predictive of prolonged survival. This analysis was conducted to assess the median number of AZA treatment cycles associated with achievement of first response, as measured by IWG 2000-defined CR, PR, or HI (major + minor). The number of treatment cycles from first response to best response was also measured.
Methods. Pts with higher-risk MDS (FAB: RAEB, RAEB-T, or CMML, and IPSS: Int-2 or High) were included. Pts were randomized to AZA (75 mg/m2/d SC × 7d q 28d) or to a conventional care regimen (CCR). AZA treatment was continued until disease progression (or unacceptable toxicity), regardless of hematologic response. Erythropoiesis stimulating agents were not allowed.
Results. In all, 358 pts were randomized (179 to AZA and 179 to CCR). Of the 179 AZA pts, 91 (51%) achieved a CR, PR, or HI. For the 91 pts who achieved an IWG response, the median number of cycles to first response was 3 (range: 1 – 22), 81% of pts achieved a first response by 6 cycles, and 90% achieved a first response by 9 cycles. For 57% of responders (n=52), their first response was their best response; the remaining 43% (n=39) had an improvement in their response status at a median of approximately 4 additional treatment cycles (range 1–11 treatment cycles) after their first response.
Conclusions. While many pts achieving a hematologic response with AZA do so in early treatment cycles, continued AZA dosing can further improve pt responses. In the AZA-001 study, a significant OS benefit was observed compared with CCR. In this study, AZA pts received a median of 9 treatment cycles (range 1–39). For those achieving a response of HI or better, 90% did so by 9 cycles; more than 40% of responders later achieved an improved response. In the absence of unacceptable toxicity or disease progression, continued AZA treatment is appropriate and may maximize patient benefit.
Identification of Genes Associated with Inhibitor Development in Hemophilia a by Microarray Analysis.
Authors: Hwang Sung Ho , Kim Hugh C , Lee Hyun Woo , Yoo Ki young , Kim Hyon J , Kim Hye Sun 
Published: April 2020

Abstract
The risk of factor VIII (FVIII) inhibitor development increases by the disease severity, presence of family history and patient-related environment. Recently, FVIII genotypes and genes related to immune response were found to be decisive risk factors for inhibitor development. To identify the differentially expressed genes (DEGs) for developing inhibitory antibodies in hemophilia A, we analyzed the gene expression profiles between inhibitor and non-inhibitor by microarray technique. The results show that the 384 genes were up-regulated and 161 genes were down-regulated in inhibitor patients as compared with non-inhibitor patients. The 545 of DEGs were classified by the functional gene grouping using Panther classification method. Of interest was the finding that the expression levels of immunity and signal transduction related genes were significantly modified in inhibitor patients. For 7 genes, validation of these bead-array data was carried out by semi-quantitative RT-PCR. Finally, we performed Real Time PCR analysis for the significantly changed DEG450. As a result, expression of DEG450 was significantly down-regulated in inhibitor patients. We demonstrate that inhibitor development is closely related to the expression levels of immune-related genes and that these genes can be used as biomarker genes for prediction of inhibitor development
Mapping of Conformational Epitopes, and Analyze of the Distribution of Ig Isotypes and Subclasses Using a Semi-Quantitative Multiplex Assay in 6 Cases of Postpartum Acquired Hemophilia.
Authors: Lavigne-Lissalde Geraldine , Lapalud Priscilla , Pouplard Claire , Granier Claude , Borg Jeanne-Yvonne , Rotschild Chantal , Gruel Yves , Schved Jean-François 
Published: April 2020

Abstract
Background and objectives: Acquired hemophilia (AH) is a severe life-threatening autoimmune disease due to the development of polyclonal autoantibodies (autoAbs) which neutralize the procoagulant activity of coagulation factor VIII (FVIII). The most common epitopes for autoAbs to FVIII are located in the A2, A3 and C2 domains of FVIII. We have recently developed a multiplex assay based on the Luminex technology that allows simultaneously the detection and epitope mapping of anti-FVIII antibodies (Lavigne-Lissalde et al, Thromb; Haemost. 2008). This technology is fast and requires only 100 μl of plasma and we showed that most autoAbs to FVIII are monospecific and preferentially bind to the light chain (LC) of FVIII. The aim of the present study was to evaluate the epitope specificity and IgG isotype/subclasses of autoAbs to FVIII detected in the post partum period.
Methods: Acquired hemophilia was diagnosed in six women (median age: 28 years) included in a French cohort (SACHA) and who exhibited prolonged activated partial thromboplastin time, low FVIII levels (range = 0.5–21 IU/dL) and inhibitor titers between 1.5 and 40 Bethesda units (BU). The epitope mapping was studied in each patient by testing 5 plasma samples collected (at inclusion and after 1, 3, 6 and 12 months of follow up) using a multiplex assay as previously described (Lavigne-Lissalde et al, Thromb; Haemost. 2008). A panel of monoclonal antibodies that bind to LC, heavy chain (HC) and the C2 domain of FVIII was employed and the use of specific immunoconjugates also allowed to define the isotype (IgG, A, M) and subclass (IgG1 to IgG4) distributions of autoAbs to FVIII. Results were expressed in mean fluorescence intensity (MFI).
Results: The autoAbs detected in the post partum in the six women studied recognized both and simultaneously HC and LC with predominant epitopes located in the LC and the C2 domain in 5 cases. At the inclusion, the autoAbs were mixtures of IgG1 and IgG4 in all patients, but the latter subclass was predominant in most cases. In addition, IgM Abs were also present in 3 women. No correlation between the titer of inhibitor (in BU) and the level of autoAbs binding (in MFI) could be evidenced. The evolution was good in all patients but one case for whom an increase in IgG4 binding to the FVIII C2 domain was demonstrated whereas inhibitor titer decreased (from 40 to 2.6 BU). Importantly, no modification in the epitope specificity between inclusion and 12 months of follow-up was found.
Conclusion: The study shows that autoAbs to FVIII developed in the context of pregnancy are mainly IgG4 specific to the C2 domain. In addition, the epitope mapping of anti-FVIII antibodies by multiplex approach could contribute to a better understanding of the pathophysiology of acquired hemophilia.
Efficacy and Safety of Rituximab in Adult Patients with Idiopathic Relapsed or Refractory Thrombotic Thrombocytopenic Purpura (TTP). Results of a Spanish Multicenter Study.
Authors: de la Rubia Javi er , Moscardo Federico , Gómez María J , Guardia Ramon , Rodriguez Pilar , Sebrango Ana , Zamora Concha , Deben Guillermo , Goterri Rosa , Lopez Rafaela , Peña Francisco , Pujol Micola 
Published: April 2020

Abstract
Background: Therapeutic plasma exchange (PE) is the mainstay of treatment of thrombotic thrombocytopenic purpura (TTP). In many patients, prolonged PE is required to achieve remission and between 30% and 60% of patients relapse after a variable period of months to years, with a further episode of TTP or a chronic picture of recurrent relapses. Various immunosuppressive agents have been used in acute refractory and chronic relapsing cases. Despite advances in treatment, mortality remains at 15–20%. Limited clinical data from small groups of patients suggest that the administration of anti-CD20 antibody (Rituximab) may also be useful in acute refractory and chronic relapsing TTP. However, whether rituximab could be and effective strategy for adults with relapsed or refractory TTP remains unresolved.
Patients and Methods: We have studied the clinical response to rituximab in 22 adult patients (16 F/6 M; median age, 43 years, range 24–72) from 11 Spanish centers who presented with acute refractory (13 cases) or an acute relapse (9 cases) episode of idiopathic TTP. Second-line therapies previously administered included vincristine (11 cases) and splenectomy (2 cases). At the time of the study, five (28%) patients presented neurological symptoms, three (17%) renal failure, and two (17%) fever. On admission, every patient received daily PE and corticosteroids during the acute episode.
Results: Rituximab was administered a median of 16 days (range, 7–53) after starting PE. It was administered immediately after PE at a dose of 375 mg/m2 each week for a median (range) of 4 (1–8) doses. No severe acute or delayed toxicity associated with rituximab was observed. Two (9%) patients died due to TTP-related causes 24 and 48 hours after relapse and were not evaluable for response. The remaining 20 (91%) patients achieved complete remission in a median of 21 days after initiating rituximab (range, 2–35). After a median follow-up of 13 months (range, 1.5–33), 17 patients are in remission and 3 patients have relapsed at 7, 29, and 29 months after rituximab administration.
Conclusion: Based on these results, rituximab appears a safe, effective, targeted therapy associated with a high response rate in adult patients with acute refractory/relapsing idiopathic TTP.
Proof of Concept for the Anti Von Willebrand Factor Aptamer in Patients with Relapsing Thrombotic Thrombocytopenic Purpura (TTP).
Authors: lma Bernd Ji , Jilma Petra , Paulinska Petra , Gilbert James C. , Hutabarat Renta , Wagner Trish , Knoebl Paul 
Published: April 2020

Abstract
Background: Thrombotic thrombocytopenic purpura (TTP) has a high morbidity and mortality rate despite current standard therapy comprising plasma exchange (PEX). Aim of this prospective clinical trial was to test the safety and efficacy of the anti von Willebrand Factor aptamer ARC1779 in patients with relapsing TTP, and to find possible proof of concept.
Methods: Three patients with relapsing TTP (with detectable anti-ADAMTS13 autoantibodies) received bolus primed continuous intravenous infusions of ARC1779 (0.002 mg/kg/min) on top of standard PEX therapy until remission of TTP was induced, or for 14 days whatever came first. ARC1779 concentrations were quantified by a highperformance liquid chromatography/ultraviolet assay, the inhibitory effect of ARC1779 on vWF activity was evaluated with an ELISA kit (READDS vWF Activity ELISA Test Kit, Corgenix, Inc, Westminster, Colo), and platelet function was assessed with the platelet function analyzer (PFA-100).
Results: ARC1779 was well tolerated without any evidence for bleeding in these patients. Median ARC1779 concentrations of approximately 10μg/mL in plasma were reached under steady state, and inhibited the collagen binding site of the vWF A1 domain by &gt;95%. While plasma exchange (PEX) removed some 50% of the drug from the blood, ARC1779 concentrations could be restored by subsequent mini boluses.
Platelet counts in all patients normalized with concurrent ARC1779 and PEX: 2 days after initiation of ARC1779 in two patients, and after 6 days in the third. In the third, the time course of response (see Figure) provides support for the concept that ARC1779 can restore platelet counts, and thus interferes with the disease process. While PEX was stopped when platelet counts reached 125/nL, 4 more days of therapy with ARC1779 increased platelet counts to 260/nL. Discontinuation of the ARC1779 infusion was associated with an immediate drop in platelet counts, so that PEX had to be restarted after 2 days (Figure).
Conclusion: These data indicate that ARC1779 effectively inhibits VWF and thereby increases platelet counts in TTP patients. Addition of ARC1779 to PEX may have the potential to accelerate recovery from organ dysfunction in TTP and thereby decrease morbidity/mortality.
Figure Figure
Graft-Versus-Host Tolerance in Dogs Following T-Cell Depleted Bone Marrow Transplantation with Absorbed ATG or CD6-Antibody.
Authors: Zorn Julia , Kolb Hans Jochem 
Published: April 2020

Abstract
Graft-versus-host disease (GvHD) is the major obstacle of allogeneic stem cell transplantation. Depletion of T-cells from the graft reduces the risk of GvHD, but results in a higher risk of leukemia relapse. Adoptive immunotherapy with donor lymphocyte transfusion (DLT) has been shown to control leukemia in patients after T-cell depleted allogeneic stem cell transplantation. However, GvHD may occur, if DLT is given too early after transplantation. In canine models of DLA-identical and DLA-haploidentical bone marrow transplantation, we compared different methods of T-cell depletion (TCD) and investigated the potential of DLT at different times after transplantation to induce GvHD. T-cell depletion was performed either with absorbed anti-thymocyte globuline (aATG) or with a combination of CD6-antibody and baby rabbit complement. ATG was absorbed with erythrocytes, liver, kidney and spleen for eliminating antibodies against stem cells. CD6-antibody (M-T606) and rabbit complement depleted T-cells effectively without affecting hematopoietic progenitor cells. Unlike aATG, monoclonal CD6-antibody spares natural killer (NK) cells and some CD8-positive cells. Treatment of bone marrow with aATG prevented GvHD in 9 dogs following DLA-identical transplantation. DLT on days 1 and 2 or 21 and 22 induced fatal GvHD in two dogs each. However, it did not induce GvHD when given on days 61 and 62 and later. In DLA-haploidentical bone marrow recipients, non-manipulated marrow produced fatal GvHD in all dogs (n=7), whereas marrow treated with aATG (vol:vol 1:100 and 1:200) produced fatal GvHD in 5 out of 16 dogs only. CD6-depletion prevented GvHD in 3 of 3 DLA-haploidentically transplanted dogs. DLT produced fatal GvHD in one dog each, when given on day 3, 7 or 14 after CD6-depleted haploidentical bone marrow transplantation. However, it produced fatal GvHD in only 2 of 4 dogs transfused on day 20 post grafting. Thus, DLT could be given earlier in DLA-haploidentical animals transplanted with CD6-depleted marrow than in DLA-identical animals transplanted with aATG treated marrow without producing GvHD. These findings support the hypothesis that graft-versus-host tolerance can be induced earlier with grafts not depleted of NK cells. NK cells in the graft may inactivate host dendritic cells necessary for the induction of GvHD. In grafts depleted with aATG, NK cells are depleted as well, because aATG still retains broad specificity despite extensive absorptions. This leaves host DCs unaffected. Transfused donor T-cells encountering this environment will thus be activated which results in severe GvHD. In contrast, monoclonal CD6-antibody spares NK cells, so that donor lymphocytes cannot be activated by host DCs at the time of DLT and thus won’t trigger GvHD. CD6-depletion is the preferred method if adoptive immunotherapy with DLT is planned.
Donor T Cell Production of IL-21 Accelerates Graft-Versus-Host Disease Lethality.
Authors: Bucher Christoph , Vogtenhuber Christine , Jasperson Lisa , Panoskaltsis-Mortari Angela , Goren Emily , Sivakumar Pallavur , Blazar Bruce R. 
Published: April 2020

Abstract
IL-21, produced by T-cells, binds to the common gamma chain family member, IL21R, expressed on immune and colonic epithelial cells. IL-21 signaling results in the maturation, activation and proliferation of T, B, NK-cells and DCs. IL-21 has been implicated in Th17 generation/amplification and also modulating Treg differentiation. However, the relevance of this in disease is unclear. Therefore, we wanted to study the effects of IL-21 depletion and the role of Th17 and Treg cells in the context of GVHD. A lethally irradiated, complete MHC disparate model (B6 to B10.BR) using donor bone marrow cells alone or with the addition of wild-type (wt) CD4+CD25- Effector T-cells and irrelevant or anti-IL-21 Ab from days 0 to 25 twice per week. The administration of anti-IL-21Ab lead to a significant delay of weight loss and mortality (P&lt;0.0001). To determine whether IL-21 deficiency accelerates or inhibits GVHD, studies were performed comparing GVHD-inducing ability of wt or IL-21 knockout (IL21−/−) donor CD4+CD25- T cells. A striking survival advantage of recipients of IL-21−/− vs wt CD4+CD25- T cells was observed (100% vs 0% surviving, P &lt; 0.0001). Subsequent studies were performed using whole T cells, a more clinically relevant donor T cell graft source. Whereas recipients of wt T-cells all died of GvHD within 50 days of GvHD, recipients of IL-21 −/− T-cells showed significantly less weight loss and superior long-term survival (P &lt; 0.0001). Histopathological examination of recipients of CD25-depleted T-cells on d14 revealed significantly reduced GvHD scores of the colonic mucosa and a trend towards lower GvHD scores in the small intestine, liver and the spleen in recipients of IL-21−/− cells. Although flow cytometry analysis of mononuclear cells showed no changes in the frequency of IL-17 producing cells in spleen, liver and colon, the frequency of interferon gamma producing CD4+ T- (Th1) cells was significantly lower in the spleen and the colon of recipients of IL-21−/− vs. wt T-cells. Moreover, increased Treg frequencies were seen in the colon of IL-21−/− vs wt CD25-depleted T cells (mean values: 7.5% vs. 2.1%; P &lt; 0.003). We conclude that IL-21 production by either CD25- depleted T-cells or CD4+CD25- T cells is sufficient to increase GVHD mortality and that Treg inhibition, rather than Th17 generation or amplification, are likely contributing to GVHD lethality acceleration. We also conclude that IL-21 neutralization represents a novel approach for GVHD inhibition that warrants further investigation.
Large Scale Selective Ex Vivo Expansion of CD4+CD25+FOXP3+ Regulatory T Cells from Peripheral Blood.
Authors: Nagamura-Inoue Tokiko , Ogami Kazuo , Yokoyama Kazuaki , Izawa Kiyoko , Kobayashi Seiichiro , Nakayama Shin , Ishige Ikuo , Inoue Yusuke , Ooi Jun , Uchimaru Kaoru , Takahashi Satoshi , Tojo Arinobu 
Published: April 2020

Abstract
CD4+CD25+Foxp3+ regulatory T cells (Treg) play an important role in allograft- and self-tolerance and thus have the potential for therapeutic application in immunological and allergic disorders. However, the frequencies of Treg in peripheral blood are very low. Here we attempted the ex vivo expansion of Treg to enable the adaptive immunoregulatory therapy in humans. CD4+ T cells from peripheral blood of healthy donors or patients with chronic graft-versus-host disease (GVHD) were isolated by anti-CD4 monoclonal antibody (MAb)-conjugated magnetic beads, and cultured using a plastic plate coated with anti-CD28 and anti-CD3-MAbs in the medium containing recombinant human (rh) IL-2 and rhTGF-b. After one week of culture, expanding cells were once detached from the plate and subjected to the fresh medium including rhIL-2 and rhTGF-b but not MAbs. After 2-weeks of culture, phenotypic and functional analyses were performed. Mixed lymphocyte reaction was done using CFSE-labeled responder T cells and autologous or allogeneic dendritic cells (DC) with or without expanded Treg-rich populations. Xenogeneic -GVHD in NOD-Scid mice was induced by the injection of human T cells expressing luciferace transgene, followed by in vivo bioluminescence imaging (BLI) analysis using a CCD camera.
Our expansion procedure with TGF-b yielded 45-83% purity of Foxp3+CD25+CD4+Treg co-expressing CTLA-4, CD54 and GITR, while 8-42% purity without TGF-b(p&lt;0.001). These cell populations also displayed CD45RO+CD45RA−CD26high+ memory phenotype. An average expansion rate of Treg was 62,200 fold (25,500–97,900) in healthy donors during the culture periods (n=5). Thus, we obtained an average of 4.7x108 Treg from the initial number of 5x105 CD4+ T cells in peripheral blood. Additionally, from peripheral CD4+ T cells in patients with chronic GVHD, Treg could be expanded equivalently to healthy donors. The resulting Treg-rich populations inhibited the proliferative response of CFSE-labeled T cells to autologous and allogeneic DC (Figure 1). The ex vivo expanded Treg-rich populations had the inhibitory effect on xeno-reactive T cells expressing luciferase transgene in a xenogeneic GVHD model (Figure 2). Our procedure has allowed efficient ex vivo expansion of Treg-rich populations from a small volume of peripheral blood, and will be applicable to clinical use.
Figure 1. MLR inhibited by expanded Treg Figure 1. MLR inhibited by expanded Treg Figure 2. Xenogeneic GVHD diminished by expanded Treg. Figure 2. Xenogeneic GVHD diminished by expanded Treg.