bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human Lymphoid and Myeloid Cell Development in NOD/LtSz-scid IL2rγnull Mice Engrafted with Mobilized Human Hematopoietic Stem Cells.
Authors: Shultz Leonard , Lyons Bonnie L. , Burzenski Lisa M. , Gott Bruce , Chen X. , Chaleff Stanley , Kotb Malak , Gillies Stephen D. , Greiner Dale L. , Handgretinger Rupert 
Published: April 2020

Abstract
We have developed, characterized, and validated a new genetic stock of IL-2r common γ (gamma) chain deficient NOD/LtSz-scid (NOD-scid IL2rγnull) mice that support high levels of human hematopoietic stem cell (HSC) engraftment and multilineage differentiation. Histology, flow cytometry, and functional assays document a severe depletion of lymphocytes and NK cells in NOD-scid IL2rγnull mice. These mice survive beyond 16 months of age and untreated as well as sub-lethally irradiated NOD-scid IL2rγnull mice are resistant to the development of lymphomas and are “non-leaky” throughout life. Intravenous injection of sub-lethally irradiated NOD-scid IL2rγnull mice with 7 x 105 human mobilized CD34+ stem cells leads to high levels of multilineage engraftment. At 10 weeks after engraftment, percentages of human hematopoietic CD45+ cells are six-fold higher in the bone marrow of NOD-scid IL2rγnull mice as compared to NOD-scid controls. Human CD45+ cells include immature and mature B cells, NK cells, myeloid cells, plasmacytoid dendritic cells and HSCs. Spleens from engrafted NOD-scid IL2rγnull mice contain high percentages of immature and mature B cells but low percentages of T cells. Treatment with human Fc-IL7 fusion protein leads to a high percentage of human CD4+CD8+ immature thymocytes and high percentages of CD4+CD8− and CD4−CD8+ mature human T cells in the spleen and blood. Validation of de novo human T cell development was carried out by quantifying T cell receptor excision circles in thymocytes and by analyses of TCRβ repertoire diversity. Human T cell function was evidenced by proliferative responses to PHA and streptococcal superantigen. NOD-scid IL2rγnull mice engrafted with human HSC generate differentiated functional human T and B cells and provide an in vivo model of multilineage human hematopoietic cell engraftment.
Bendamustine/Mitoxantrone/Rituximab (BMR): A Very Effective and Well Tolerated Immuno-Chemotherapy for Relapsed and Refractory Indolent Lymphomas - Results of a Multicentre Phase-II-Study of the German Low Grade Lymphoma Study Group (GLSG).
Authors: Weide Rudolf , Koeppler Hubert , Heymanns Jochen , Thomalla Joerg , Hess Georg , Gamm Heinhold , Aldaoud Ali , Schmitz Stephan , Haak Ursula , Plewe Dirk , Burkhard Oswald , Gaede Bernd , Hinrichs Fokke , Pflueger Karl-Heinz , Dreyling Martin , Unterhalt Michael , Hiddemann Wolfgang 
Published: April 2020

Abstract
In a previous phase-I-study we could demonstrate that the combination of the two chemotherapeutic agents bendamustine and mitoxantrone in combination with the lymphocyte-specific antibody rituximab (BMR) is a highly effective regimen in the treatment of relapsed or refractory indolent lymphomas and CLL (Leukemia and Lymphoma 2002, 43(2):327–331). Based on these data we have conducted a multicenter phase-II-study to further evaluate the efficacy and toxicity of BMR. BMR treatment schedule consisted of Bendamustine 90mg/m2 (days 1+2), Mitoxantrone 10mg/m2 (day 1) and Rituximab 375mg/m2 (day 8). Treatment was repeated on day 29 for a total of 4 cycles. Between 04/03 and 07/04, 39 patients (pts) with symptomatic stage III/IV indolent lymphomas were treated with BMR (18 follicular, 11 mantle cell, 3 marginal zone, 6 immunocytoma and 1 patient with hairy cell leukemia). All pts were treated on an outpatient base. Median age was 67 years (40 – 83) and their performance score ranged from 0–2. Median number of previous treatment regimens was 1 (1–8). 16 pts (41%) had rituximab pretreatment. Currently, 25 pts are evaluable: A reversible grade 3/4 hematotoxicity occurred in 19 pts (76%), no treatment related death was reported. The overall response rate was 92% with 36% complete remissions and 56% partial remissions. Updated results on progression-free and overall survival rates will be presented. We conclude that BMR is a very effective and well tolerated immuno-chemotherapy for relapsed or refractory indolent lymphomas.
Blockade of p110delta Isoform Activity of Phosphoinositide 3-Kinase Inhibits Blast Cell Proliferation in Acute Myeloblastic Leukemia.
Authors: Bouscary D. , Bardet V. , Sujobert P. , Cornillet-Lefebvre P. , Hayflick J. S. , Prie N. , Verdier F. , Vanhaesebroeck B. , Muller O. , Pesce F. , Ifrah N. , Berger M. Hunault , Bertho C. , Guillerm G. , Villemagne B. , Audhui B. , Solary E. , Witz F. , Harousseau J. L. , Himberlin C. , Lamy T. , Lioure B. , Bene M. C. , Cahn J. Y. , Dreyfus F. , Mayeux P. , Lacombe C. 
Published: April 2020

Abstract
AMLs constitute an heterogeneous group of hematopoietic stem cell disorders of rather poor prognosis. Abnormal activation of signaling pathways including STAT, MAPK and NF-KB have been described in AMLs and could constitute new targets for therapy. In this study, we focused on the PI3K/AKT and mTOR/P70S6K activity in primary blast cells purified from the bone marrow of patients with primary (n = 54) or secondary (n =10) AMLs. Patients with de novo AMLs were issued from the AML2001 French Multicenter Trial directed by the GOELAMS. The FAB classification was the following: M0 (n = 8), M1 (n = 10) M2 (n = 21), M4 (n = 10), M4eo (n = 2), M5 (n = 11), unknown AML (n = 2). In 58% of these patients, a constitutive activation of PI3K/AKT was detected in blast cells by western blot analysis showing AKT and FOXO3a phosphorylation, and confirmed by immunofluorescence confocal microscopy and flow cytometry analysis in CD34+ blast cells. Constitutive GAB1/2 phosphorylation was detected in all PI3K+ patients. However, we did not find any correlation between PI3K activation and FLT3 gene mutations. PTEN and SHIP-1 expression was normal in all tested PI3K+ patients. Proliferation (3H thymidine incorporation) was significantly increased in PI3K+ samples compared to PI3K- samples (p=0.001). The mTOR/P70S6K pathway was activated and the mTOR inhibitor rapamycin selectively reduced proliferation of PI3K+ samples. We determined the expression of the alpha, beta and delta isoforms of the catalytic sub-unit of PI3K (p110) in leukemic cells and found that p110 delta was the only consistently expressed isoform. In 8 PI3K+ samples tested, IC87114 (ICOS Corporation), a specific p110delta inhibitor compound used at 10μM totally suppressed AKT and FOXO3a phosphorylation. IC87114 inhibited proliferation of PI3K+ leukemic cells whereas it did not induce apoptosis. IC87114 did not significantly impaired the proliferation of PI3K- blast cells. Interestingly, IC87114 did not inhibit the proliferation or clonogenicity of CD34+ cord blood cells cultured in SCF, FLT3L and TPO used as controls.
Overall, our results report, a/ a constitutive activation of PI3K in 58% of AML, b/ no correlation between FLT3 gene mutations and PI3K positivity, c/ no implication of PTEN or SHIP-1 phosphatases in PI3K activation, d/ a major role of the p110 delta isoform of PI3K in leukemic cell proliferation, e/ a potential therapeutic value of inhibiting mTOR and P110 delta activity.
EB10, an Anti-FLT3 Monoclonal Antibody, Prolongs Survival and Reduces NOD/SCID Engraftment of AML Cell Lines and Primary Blasts.
Authors: Piloto Obdulio , Levis Mark , Li Li , Nguyen Bao , Kim Kyu-Tae , Huso David , Li Yiwen , Li Hongli , Wang Mei-Nai , Witte Larry , Hicklin Daniel J. , Small Donald 
Published: April 2020

Abstract
The FLT3 receptor is a potential target in AML due to its role in leukemogenesis and its high degree of expression on blasts from approximately 90% of acute myeloid leukemia (AML) patients. In addition, mutant forms of FLT3, including internal tandem duplications (ITD) in the juxtamembrane region and point mutations in the kinase domain, constitutively activate FLT3 signaling. ITD mutations in particular are also associated with poor prognosis. A number of small molecule tyrosine kinase inhibitors (TKI) against FLT3 are currently in clinical trials and have shown some clinical activity. However, TKIs have various limitations, including their lack of specificity, which may produce toxicities, and can select for drug resistant cells. In an attempt to overcome some of these limitations and to generate new agents which might cooperate in targeting FLT3, we generated a fully humanized phage display monoclonal antibody (EB10). This antibody is capable of inhibiting both ligand-activated wild-type and, to a lesser degree, ligand-independent mutant FLT3 signaling. When EB10 is used to treat cells expressing activated FLT3, inhibition of downstream pathways including STAT5, AKT and MAPK are also frequently seen. EB10 treatment of cells expressing FLT3 in the presence of NK cells leads to antibody-dependent cell-mediated cytotoxicity (ADCC). EB10 treatment of NOD/SCID mice injected with FLT3 expressing AML cell lines or with primary AML blasts significantly prolongs survival and/or reduces engraftment of leukemic cells. EB10 proved efficacious in vivo against cells even when in vitro EB10 treatment did not significantly reduce FLT3 signaling. This indicates that ADCC may be the primary mechanism mediating cytotoxicity as opposed to direct FLT3 inhibition. In contrast to the effects on AML cell lines and primary samples, EB10 treatment did not significantly reduce NOD/SCID engraftment of normal human CD34+ hematopoietic stem cells. Anti-FLT3 antibodies, like EB10, may be a promising therapeutic agent that can specifically target malignant cells with limited toxicities against normal hematopoietic stem cells and should be considered for clinical trials.
Reduced Plasma Fibrinolytic Potential Is a Risk Factor for Venous Thrombosis.
Authors: Lisman Ton , de Groot Philip G. , Adelmeijer Jelle , Meijers Joost C.M. , Rosendaal Frits R. 
Published: April 2020

Abstract
The role of the fibrinolytic system in the development of deep vein thrombosis (DVT) is unclear. We determined the plasma fibrinolytic potential of patients enrolled in the Leiden Thrombophilia Study (LETS), a population-based case-control study on risk factors for DVT. Plasma fibrinolytic potential was determined in 421 patients and 469 controls who were not using oral anticoagulants at the time of the blood draw by means of a tissue factor- and tissue-type plasminogen activator (tPA)-induced clot lysis assay. This assay represents an overall measure for plasma fibrinolytic potential, as it was previously shown to be dependent on plasma levels of plasminogen, α2-antiplasmin, plasminogen activator inhibitor type 1, and thrombin activatable fibrinolysis inhibitor. Using clot lysis times above the 70th, 80th, 90th, 95th, and 99th percentile of the values found in control subjects as cut-off levels, we found a dose-dependent increase in risk for DVT in patients with hypofibrinolysis (odds ratio’s [95 % CI] of 1.4 [1.1–1.8], 1.6 [1.2–2.1], 1.9 [1.3–2.9], 2.1 [1.3–3.6], 2.2 [0.7–7.5], respectively). This indicates a two-fold increased risk of DVT in subjects with clot lysis times above the 90th percentile. Using the 90th percentile, we showed that the risk was not affected by age or sex (odds ratio [95% CI] 2.0 [1.3–3.0]), and after correction for all possible confounders (age, sex, and levels of procoagulant proteins which were shown to associate with clot lysis times in the control population) the risk estimate was marginally reduced (odds ratio [95% CI] 1.6 [1.0–2.5]). Taken together, these results indicate that plasma hypofibrinolysis constitutes a risk factor for venous thrombosis. Whether this hypofibrinolytic state is determined by genetic or acquired factors, or a combination of these, and which proteins are involved is at present unknown.
Economic Evaluation of Bortezomib (VELCADE) for Relapsed and Refractory Multiple Myeloma.
Authors: Bagust Adrian , Haycox Alan R. , Boland Angela , Chu Patrick , Morgan Gareth J. , Mujica Mota Ruben E. , Dubois Dominique , Dhawan Ravinder 
Published: April 2020

Abstract
Bortezomib (VELCADE), a new proteasome inhibitor, was recently licensed by the FDA for treating patients with relapsed and refractory multiple myeloma (MM), and has now also been approved for similar use in Europe. A model was developed to evaluate and compare the costs and health benefits of bortezomib, as a third-line treatment for patients with relapsed and refractory MM, relative to best supportive care in the UK. A two-part mathematical model of survival was built and calibrated using individual patient data from the SUMMIT trial (n=202 ; NEJM2003;348:2609-17) , a multi-center, pivotal phase II trial of bortezomib in 202 patients with relapsed and refractory MM. Patients in SUMMIT were 18 years of age or older, with a life expectancy of more than three months and a Karnofsky score &gt;=60. In the first part of the survival model, the time to disease progression for patients on bortezomib (n=202) or best supportive care (n=38) was estimated. In the second part, the time from disease progression till death was calculated, based on the sub-sample of patients for whom relevant data were available (n=79). Health-related utility was assessed for the effects of the drug and also the natural course of the disease on quality of life, yielding different utility weights pre- and post-disease progression. Those values were derived by mapping individual patient reports of quality of life in SUMMIT (EORTC-QLQ30 and MY24, FACIT-Fatigue and FACT/GOG-Neurotoxicity subscales) onto the 5 scales of the EQ-5D instrument. Resource use data from SUMMIT were used to estimate costs from the perspective of the UK NHS for bortezomib administration, hospital care, medications and diagnostic tests and surgical procedures on an individual patient basis. Results: By delaying the rate at which disease progresses, bortezomib produced survival gains relative to best supportive care that range between 7.75 to 12.09 months of life depending on the method of survival estimation. Additional total direct costs associated with the novel agent were £17290 or £24121 if the additional costs incurred during the extended period of survival are included. The incremental cost-effectiveness ratio for bortezomib was in the range £17161- £33539 per life year gained and £26714 - £51,666 per quality-adjusted life year gained. Conclusions: These results suggest that bortezmib is a cost-effective third-line treatment option for patients with RRMM in the UK. This study provides a methodological framework for undertaking preliminary economic evaluations of new drugs for the treatment of cancer with the aim of informing purchasing decisions based on limited outcome data.
Non-Hematopoietic Bone Marrow Cells Can Be Mobilized and Differentiate into Cardiomyocytes after Myocardial Infarction: Possible Contribution of Mesenchymal Stem Cells.
Authors: Kawada Hiroshi , Fujita Jun , Kinjo Kentaro , Matsuzaki Yumi , Tsuma Mitsuyo , Miyatake Hiroko , Muguruma Yukari , Okano Hideyuki , Hotta Tomomitsu , Fukuda Keiichi , Ando Kiyoshi 
Published: April 2020

Abstract
Recent studies have suggested that bone marrow (BM) cells can contribute to regeneration processes in various tissues. Cardiomyocytes derived from BM cells have been observed after myocardial infarction (MI), and BM-derived cells mobilized by cytokines were capable of regenerating the myocardial tissue, leading to an improvement in survival or cardiac function after MI. However, the responsible BM cells have not been fully identified. The most likely candidates for the BM-derived stem cells with the ability to regenerate myocardial tissue are hematopoietic stem cells (HSC) and mesenchymal stem cells (MSC). The aim of this study was to determine the precise origin of the BM cells mobilized by cytokines to repair infarcted myocardium. Here, we used two independent clonal studies; we transplanted genetically marked single HSC or clonal MSC into lethally-irradiated recipient mice, induced MI, treated the mice with G-CSF, and analyzed the cardiac tissues. First, we transplanted single CD34−c-kit+Sca-1+lineage− side population (CD34−KSL-SP) cells or whole BM cells from mice ubiquitously expressing enhanced green fluorescent protein (EGFP) into lethally-irradiated mice, induced myocardial infarction (MI), and treated the animals with G-CSF to mobilize stem cells to the damaged myocardium. At 8 weeks post-MI, from 100 specimens we counted only 3 EGFP+actinin+ cells in myocardium of CD34−KSL-SP cells-transplanted mice, but more than 5,000 EGFP+actinin+ cells in whole BM cell-transplanted mice, suggesting that most of EGFP+actinin+ cells derived from non-hematopoietic BM cells. These results suggested that the major population of cells mobilized from the BM and active in the regeneration of cardiomyocytes was non-hematopoietic in origin. Next, clonally purified mesenchymal stem cells, CMG cells, transfected with a pMLC2v-EGFP plasmid encoding EGFP driven by the myosin light chain promoter were transplanted directly into BM of lethally-irradiated mice, MI was induced, and they were treated with G-CSF. As a result, a number of EGFP+actinin+ cells were observed in the ischemic myocardium, indicating that CMG cells had been mobilized and differentiated into cardiomyocytes. Together, these results suggested that the vast majority of BM-derived cardiomyocytes are of mesenchymal origin.
Thrombotic Events in Adults with Acute Lymphoblastic Leukemia.
Authors: Cserti Christine M. , Brandwein Joseph , Lipton Jeffrey H. , McLeod Anne G. , Messner Hans , Minden Mark D. , Schimmer Aaron D. , Schuh Andre C. , Seki Jack T. , Yeo Erik L. , Yi Qi-Long , Wells Richard A. 
Published: April 2020

Abstract
A strong link between asparaginase-containing (ase +) chemotherapy regimens and venous thrombotic events (VTE) has been established for pediatric acute lymphoblastic leukemia (ALL), but this association has not been studied in adult ALL. We recently adopted as our first-line therapy for adult ALL a treatment protocol (DFCI 00–01) that employs weekly high-dose ase throughout a 27 week intensification phase. We reviewed the records of 92 adult patients consecutively accrued at Princess Margaret Hospital (from Jan 2000 to Dec 2003) for venous thrombotic events (VTE). Median age at diagnosis of ALL was 41±17 y, or 19±3 y for those who presented after a history of pediatric ALL (n=6), and 42±16 y for adults who presented with de novo ALL (n=86). Thirty-five (37%) were female, 59 (63%) male. Twenty-four (26%) were Philadelphia-chromosome positive; 71% of these received regimens containing imatinib. At least 1 symptomatic, radiologically confirmed VTE was seen in 19 patients (21%), with 3 (3%) diagnosed prior to any antileukemic therapy. Overall survival (OS) versus thrombosis-free survival (TFS) is illustrated in the Kaplan-Meier curve below. Median time to VTE was 94±92 d [95% CI]. Average age of patients with VTE was 44 ±15y; no events were observed in patients with a prior pediatric history of ALL. The sites of VTE were: 10/22 (46%) in the lower extremities, 5/22 (23%) propagating from central venous access catheters (CVAC), 4/22 (18%) as pulmonary emboli (2 in isolation), and 3/22 (14%) CNS (2 cerebral venous sinus and 1 retinal venous thrombus). The first antileukemic regimen administered was DFCI 00–01 ± imatinib in 79/92 (86%), HyperCVAD ± imatinib in 7 (8%), an unclassified ase+ regimen in 1 (1%), or an unclassified asparaginase-excluding (ase−) regimen in 5 (5%). A significant difference in the rate of ase-dependent VTE was observed when all regimens (1° or subsequent) were counted, with 18 VTE during 103 ase+ regimens (17.5%), versus only 1 VTE in 32 ase− regimens (3.1%), [p=0.04]. Median time to VTE in DFCI 00-01was 109±98d, most often occurring during the intensification phase. The odds ratio for VTE with any T-cell subtype of ALL compared to any B-cell subtype was 4.6 [p=0.0043].
Conclusions : A high, significant VTE rate (~18%) was observed in adults with ALL undergoing ase+ antileukemic therapy, nearly 6X the rate observed either prior to treatment or during ase− regimens (~3%). Ase is thus strongly associated with VTE, warranting improved surveillance and antithrombotic prophylaxis. Risk factors (underlying thrombophilias, leukemic subtypes) similarly deserve further study.
Figure Figure
Survivin-Driven Therapy for Leukemia.
Authors: Caldas Hugo , Altura Rachel A. 
Published: April 2020

Abstract
Survivin, a member of the inhibitor of apoptosis (IAP) family of proteins, is widely expressed in transformed cell lines and in many different primary cancer cells, including both hematopoietic and non-hematopoietic malignancies. It is not expressed in many non-malignant adult tissues, but is essential for fetal development, as demonstrated by conventional gene-targeting experiments in mice that show embryonic lethality at day 4–6 of development. In adult cancers, including lymphoma and many epithelial carcinomas (colon, breast, gastric) the expression level of survivin, as assayed by immunohistochemical analysis and RT-PCR, correlates with overall survival.
We have designed a novel gene therapy approach that takes advantage of the high expression levels of survivin in malignant cells, in which the survivin promoter is used to drive the expression of a suicide gene to kill cancer cells by programmed cell death. Our system is based on perforin-independent granzyme B cytotoxicity and therefore does not require pro-drug activation. We designated it SAGA, for survivin and granzyme apoptosis. We used Jurkat cells as an in vitro model for T-cell leukemia, and 697Bcl2 cells as a model for pre-B Bcl2-expressing leukemia, to show this approach is more efficient in killing leukemic cells than conventional chemotherapy. Jurkat cells responded to both vincristine therapy and SAGA whereas 697Bcl2 were unaffected by vincristine, but responded to SAGA. Cell growth curves of Jurkat cells and 697Bcl2 cells are shown in Figure 1. Our approach not only inhibits cell growth, but also induces apoptosis. We detected apoptotic events by Annexin V staining and by changes in mitochondrial potential, as early as 12 hours post-treatment. Rates of early apoptotic events are shown in Table 1. In addition to these events, we also documented DNA fragmentation and caspase-3 activation in treated cells. Cytotoxicity was clearly visible by microscopic analysis 24 hours post-treatment (Figure 2). Our results strongly suggest that survivin-driven suicide gene therapy effectively enhances cell death of leukemic blasts derived from two common sub-types of ALL, one of which expresses the potent anti-apoptotic inhibitor, Bcl-2, known to be clinically more resistant to standard therapy. Experiments evaluating the in vivo effects of SAGA in mouse leukemia models are currently underway.
Cell death at 24 hours
Early Apoptosis Necrosis control vincristine SAGA control vincristine SAGA T-ALL 2% 46% 41% 2% 18% 19% B-ALL 1% 2% 29% 0.4% 1% 30%
Figure Figure Figure Figure