bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Aurora Kinase Inhibitor AZD1152 Causes Perturbation of Cell Cycle Distribution in Cell Lines and Primary AML Samples.
Authors: Walsby Elisabeth , Walsh Val , Pepper Chris , Mills Ken , Burnett Alan K. 
Published: April 2020

Abstract
Aurora kinases (AK) participate in chromosome separation during mitosis and are essential for mitotic processes and completion of cytokinesis. AK-A is found at the centrosome and spindle apparatus during prophase to telophase, while AK-B is seen in the midzone during anaphase of the cell cycle. Inhibition of these kinases may be a viable therapeutic strategy. AZD1152 is a specific AK inhibitor, designed to target cell division in proliferating tumour cells, with potential for activity in a wide range of tumours. In order to evaluate the potential utility of the AK inhibitor AZD1152 in acute myeloid leukaemia (AML), the expression levels of AK-A, -B, and -C were evaluated in a panel of myeloid cell lines (HL-60, NB4, NB4R2, U937, KG1, and K562) and in cells from 10 normal bone marrows and 240 AML patients using the Affymetrix gene expression system (Affymetrix Inc.) validated by real-time quantitative polymerase chain reaction in 105 patients. AK-C was absent in all samples, while AK-A and -B were present in 5% and 34%, respectively, of the AML patients and both were present in normal bone marrow samples. Expression levels did not correlate with age, sex, French-American-British classification or cytogenetic risk group. AML cell lines were incubated with a variable concentration of AK inhibitor (0.01 μM to 10 μM) for up to 72 h and the effects on viability, cell growth, cell cycle, and DNA content measured after 24 and 48 h exposure. This resulted in the accumulation of cells with 4N DNA content, indicating an increased proportion of cells in G2, and development of cells with DNA content greater than G2/M phase cells, on occasions showing 8N ploidy. This effect was time and dose dependent. At 24 h AK inhibitor caused a significant increase (p≤0.05) in the number of cells with 4N DNA content, suggestive of G2, or with greater than 4N DNA content at concentrations as low as 0.01 μM. Growth of HL-60 and NB4 cells was inhibited, following 72 h treatment with AK inhibitor (0.01 μM), by 50% and 90% (p&lt;0.05), respectively. Incubation of AML samples (n=28) with AK inhibitor also showed an effect on the cell cycle, with a significant increase in G2 population (p≤0.05), and occasionally an increase of DNA content (p≤0.05) after 24 and 48 h at a minimum concentration of 1.0 μM. Flow cytometric analysis showed histone H3 phosphorylation was decreased, from around 3–5% in untreated cells, only in a proportion (~35%) of primary samples. The growth inhibition effects seen are combined with increased DNA content, which is consistent with the endoreduplication effects associated with these agents. AK-B is expressed in around one-third of AML patients, and agents targeted against this kinase may have therapeutic potential in AML.
Expression of Vascular Endothelial-Cadherin on Leukemic Cells Mediates Their Interaction with Vascular Endothelium.
Authors: Ellis Leslie R. , Vincent Loic , Shmelkov Sergey , Hooper Andrea , Avecilla Scott , Young Lauren , Rafii Shahin 
Published: April 2020

Abstract
It has already been established that subsets of leukemic cells express receptors for pro-angiogenic factors, such as vascular endothelial growth factor receptor-2 (VEGFR-2). Further studies have shown that these same leukemic cells also produce the ligand for these VEGF receptors, VEGF-A. This autocrine loop supports the invasion and proliferation of these particular leukemic cells. The VEGFR-2 signaling pathway is further dependent upon the co-activation of other pro-angiogenic factors, such as vascular endothelial (VE)-cadherin. VE-cadherin is an endothelial cell-specific transmembrane cellular adhesion protein that when bound results in the dephosphorylation of VEGFR-2 and contributes to neo-vessel formation. Recent studies have suggested that VE-cadherin may be expressed by a unique subset of hematopoietic cells, raising the possibility that leukemic cells may express VE-cadherin as well. We therefore sought to identify the expression of VE-cadherin on leukemic cell lines and primary samples, and further determine its role in the interaction with VE-cadherin-positive endothelial cells. Leukemic cell lines and primary leukemias were screened for the presence of VE-cadherin by both RT-PCR and Western blot analysis. Primary leukemic samples, as well as established cell lines for human erythroblastic leukemia (HEL) and acute myelogenous leukemia (KG-1a) were found to express VE-cadherin. VE-cadherin expression was further confirmed by flow cytometry and immunocytochemistry, which demonstrated that approximately 15% of the total population was VE-cadherin-positive. Proliferation and migration assays utilizing neutralizing monoclonal antibodies to VE-cadherin were performed with no obvious effects. However, immunofluorescent staining of a co-culture performed with the above leukemic cells grown on a layer of human umbilical vein endothelial cells (HUVECs) demonstrated that the leukemic cells and HUVECs interact via VE-cadherin. Furthermore, when these leukemic cells were injected into NOD-SCID mice subcutaneously, the VE-cadherin-positive cells localized around the vessels present within the leukemic chloroma. These data set forth the concept that a subset of leukemic cells expresses the protein VE-cadherin and that VE-cadherin is involved in the cell-to-cell interaction between a subset of leukemic cells and vascular endothelial cells. Heterotypic VE-cadherin interaction between leukemic cells and neo-vessels may increase the survival of leukemic cells and contribute to the generation of minimal residual disease. Therefore, inhibition of VEGFR-2 in conjunction with VE-cadherin may provide a novel strategy to eradicate minimal residual disease.
Advanced Glycation End Products (AGEs)-Mediated Cell Growth of Human Acute Myelogenous Leukemia Via Mitogen-Activated Protein (MAP) Kinase Pathways.
Authors: Kim Ju Young , Park Hyun Ki , Yoon Jin Sun , Kim Eun Shil , Ahn Kwang Sung , Yoon Sung Soo , Kim Byoung Kook , Lee Young 
Published: April 2020

Abstract
Advanced glycation end products (AGEs) are products of non-enzymatic glycation/oxidation of proteins/lipids that accumulate slowly during natural aging and at a much accelerated rate in a variety of disorders such as diabetes, renal failure, and Alzheimer’s disease. AGE modifications do not only change the physicochemical properties of the afflicted molecules, but also induce cellular signaling, activation of transcription factors and subsequent gene expression in vitro and in vivo. Most of the biologic activities associated with AGEs have been transduced by receptor for AGE (RAGE). Recently, AGEs are known to be in association with diverse cancers in terms of cellular proliferation and metastasis. However, little is known about the role of AGEs in acute myelogenous leukemia (AML). Here we examined the effects of the AGEs-RAGE interaction on the cell proliferation and intracellular signaling of AGEs in human leukemia cell lines. Expression of RAGE was observed in 8 AML cell lines examined, and up-regulated by treatment of AGE. AGE induced the proliferation of AML cell lines, HL60 and HEL, in a dose-dependent manner. Treatment with 5 μM of antisense S-ODN for RAGE did effectively inhibit cell growth of HEL cells. Exposure of HL60 and HEL with AGE induced a significant increase in the numbers of cells in S phase of cell cycle in a dose-dependent manner. AGE enhanced the expression of cell cycle regulatory proteins such as cyclin-dependent kinase (CDK) 2/4/6, cyclin D1/E/B in a dose- and a time-dependent manner. In addition, the protein levels of the cyclin-dependent kinase inhibitor (CDKI), p21 and p27, were decreased by 24 hr exposure of AGE from 10 to 200 μg/ml in HEL. Furthermore, treatment of HEL with 200 μg/ml of AGE triggered activation of mitogen-activated protein (MAP) kinases, Erk, Akt, and p38, pathways and in nuclear translocation of transcription factors NF-kB. These results indicated that AGE induced the cell growth of human AML cells, HL60 and HEL, via augmentation of cell cycle and activation of MAPK kinase pathways. Up-regulation of RAGE by exposure of AGE suggested that cellular proliferation of AML cells might be mediated in autocrine fashion.
Regulation of T Cell Homeostasis and Cell Cycling in Patients with Acute Myeloid Leukemia.
Authors: Hunger J. Katrin , Pfoertner Susanne , Ivanyi Philipp , Krauter Juergen , Ganser Arnold , Buer Jan , Franzke Anke 
Published: April 2020

Abstract
Background and Aims: Although leukemic cells express tumor-associated antigens, suppression of an effective T cell immune response is a well known phenomenon in acute myeloid leukemia (AML). In principle, the malignant cells may escape immune surveillance by (1) incomplete T cell activation due to the absence of costimulatory molecules on the leukemic cell types, (2) secretion of soluble immunosuppressive factors, which may lead among others to (3) induction of anergic and/or regulatory T cell phenotypes. A better understanding of the underlying mechanism would have important implications for the development of immunotherapeutic strategies.
Methods: CD3+ T cells were analyzed for activation, proliferation and Th1 cytokine production after coculturing with primary AML cells or leukemic cell supernatant. In addition, self-developed human Treg microarrays were used to reveal a gene expression profile of CD4+ T cells isolated from untreated AML patients (n=7) in comparison to CD4+ T cells from healthy volunteers (n=6) and AML patients after achieving a complete remission (n=4). The specific regulation of selected canditate genes was confirmed by realtime RT-PCR. Based on microarray results cell-cycle analyses were performed in order to study the effect of primary AML cells on the cell-cycle progression of stimulated T-cells.
Results: Our studies could show that primary AML cells suppress T cell activation, proliferation and Th1 cytokines production mainly in a cell-contact dependent manner and to a minor degree via soluble factors. Nevertheless, cell-cycle analyses demonstrated that soluble factors from leukemic cell supernatants may inhibit the cell-cycle entry of T cells after mitogenic stimulation with an arrest in G0/G1 phase. Interestingly, the molecular profile of CD4+ T cells isolated from untreated AML patients could identify a profound dysregulation of molecules involved in the regulation of cell-cycle (i.e., CDK4, Vav2, Egr-1, Id2) and apoptosis (i.e., Bcl-2, Bax). In addition, genes important for the immune response (i.e., IL2R, CTLA-4), especially various chemokines/-receptors (i.e., RANTES, CXCL10, CCR2) were dysregulated, whereas the expression pattern of genes specific for regulatory T cells (i.e., FoxP3, PD-1, GITR) were broadly unaffected.
Conclusion: AML cells (in-)/directly modulate normal T cell function such as activation, proliferation, cytokine/chemokine production and cell-cycle regulation. One might speculate that the lack of an effective anti-tumor response is associated with a cell-cycle arrest resulting in T cell anergy. Cell-cycle regulators may serve as potential targets for novel immunotherapeutic strategies to eradicate minimal residual disease in patients with AML.
4E-Binding Protein 1 (4E-BP1) in CD34+ Cells as Measured by Quantitative Flow Cytometry Is an Independent Prognostic Factor in Patients with Acute Myeloid Leukemia.
Authors: Gorre M. , Jilani I. , Chang R. , Bareng J. , Chan H. , Keating M. , Kantarjian H. , Estey E. , O’Brien S. , Giles F. , Albitar M. 
Published: April 2020

Abstract
The mammalian target of rapamycin (mTOR) is a serine/threonine kinase involved in the regulation of cell growth and proliferation. Once activated, mTOR can phosphorylate its downstream targets. One of these targets is the 4E-binding protein 1 (4E-BP1), which is phosphorylated and inactivated by mTOR in response to a growth signal. Phospho-4E-BP1 dissociates from the eukaryotic initiation factor 4E (eIF-4E), a translation initiation factor that subsequently binds the cap structure of 5′ mRNAs and initiates the translation of transcripts encoding genes involved in cell cycle control. Rapamycin and its analogs are immunosuppressant drugs that exert their activity by specific inhibition of mTOR. mTOR inhibition induces cell cycle arrest not only in normal lymphocytes but also in malignant cells. Using flow cytometry, we quantified the levels of 4E-BP1 and phosopho-4E-BP1 in CD34+ and CD3+ cells from bone marrow samples collected from patients with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS). We then measured the antibody binding capacity of 100 CD34+ or CD3+ cells using QuantiBRITE and PE (phycoerythrin)-labeled antibodies in a 1:1 ratio. We demonstrated that CD34+ cells express significantly higher levels of 4E-BP1 (P=0.005) and phosphor-4E-BP1 (P=0.0001) as compared with CD3+ cells in patients with AML (n=49). In contrast, there was no significant difference in the levels of 4E-BP1 or phospho-4E-BP1 between CD34+ and CD3+ cells in patients with MDS (n=15). More importantly, in patients with AML, high expression of 4E-BP1 in CD34+ cells was associated with shorter survival (P=0.003) as well as shorter complete remission duration (CRD) (P=0.03). This association between survival and levels of 4E-BP1 was independent of cytogenetic abnormalities in this group of patients. This data not only suggests that the 4E-BP1 level, as measured in the CD34+ cells, can be an important prognostic indicator in AML but also suggests that 4E-BP1 plays a role in the biology of AML. Furthermore, targeting 4E-BP1 by mTOR inhibitors, or other means of down modulating 4E-BP1 levels, is a rational therapeutic approach in AML.
Figure Figure
Tipifarnib (Zarnestra, R115777) as Maintenance Therapy for Adults in Complete Remission (CR) Following Induction and Consolidation Therapies for Poor-Risk Acute Myelogenous Leukemia (AML): A Phase II Trial.
Authors: Karp Judith E. , Gojo Ivana , Greer Jacqueline , Smith B. Douglas , Klein Maureen , Lancet Jeffery E. , Gore Steven D. , Morris Lawrence , Thibault Alain , Rybak Mary Ellen , Wright John J. 
Published: April 2020

Abstract
Disease-free survival (DFS) is short in patients ≥ age 60 or with secondary AML, adverse cytogenetics, or leukostasis at presentation. In such patients, median CR duration is ≤ 8 mos and only 20% achieve 1 yr DFS. Historically, maintenance therapy with low doses of cytotoxic chemotherapy has not prolonged DFS. We tested the hypothesis that Tipifarnib (T) might be active as maintenance therapy for adults with poor-risk AML in first CR after induction and consolidation therapies. Oral T 400 mg bid for 2/3 wks was begun median 2.4 mos (range 1.0–4.1) after start of final consolidation cycle and given for up to 48 wks (16 cycles) to 36 adults with median age 63 (range 27–82), secondary AML 31%, adverse cytogenetics 47%, leukostasis 17%, ≥ 2 risk factors 40%. T was well-tolerated, with only 4 of 36 unable to complete 2 cycles because of constitutional symptoms (1 rash, 3 non-compliant). To date, 256 cycles have been administered (median 8 per pt, range 1–16), with hospitalization required during only 4 (1.5%) cycles (infection 3, bowel obstruction 1). Dose reductions for myelosuppression (400 mg bid to 300 mg bid) occurred in 17/32 (53%) by cycle 3, and 2 (6%) needed platelet transfusions. A total of 9 patients completed all planned 16 cycles of T with a median CR duration of 24 mos (range 15–36+). Five of the 9 remain in continuous CR (CCR) 19+-36+ mos, median 26+), compared with 4 who relapsed after CR of 15–24 mos (median 22). There are 8 additional pts in CCR and still receiving T (2–12 cycles) with CCR 4+-12+ mos. A total of 15 patients progressed while on T at median 6.5 mos CR (range 3.5–12). Median CR duration for all patients is 10+ mos (range 3.5–36+), with 88% ≥ 6 mos and 48% ≥ 12 mos. In 13 “comparable” poor-risk pts (age ≥ 60 46%, secondary AML 25%, adverse cytogenetics 46%, leukostasis 23%) who were eligible for but declined T, 4 are in unmaintained CCR at 14+-25+ mos, 9 have relapsed at median 7.5 mos (5–13 mos). Treatment with T did not have a negative impact on reinduction chemotherapy at relapse, as 6 of 9 patients achieved second CR. Administration of T in CR after induction and consolidation therapy has low toxicity and is associated with prolonged DFS in some adults with poor-risk AML. Phase 3 studies are warranted in this patient population.
SU11657, a FLT3-Targeted Tyrosine Kinase, Has Pro-Apoptotic Activity on Leukemia Cells In Vitro.
Authors: Grafone Tiziana , Ferraretti Laura , Ottaviani Emanuela , Mancini Manuela , Palmisano Michela , Malagola Michele , Terragna Carolina , Renzulli Matteo , Poerio Angela , Amabile Marilina , Soverini Simona , Colarossi Sabrina , Iacobucci Ilaria , Baccarani Michele , Martinelli Giovanni 
Published: April 2020

Abstract
Fms-related tyrosine kinase3 (Flt3) is the most commonly mutated gene in human acute myeloid leukemia (AML) and has implicated in its pathogenesis. Constitutive activation of the Flt3 receptor tyrosine kinase, have been linked either by internal tandem duplication (ITD) of the juxtamembrane region or by point mutation in the second tyrosine kinase domain (TKD). To investigate the effect in vitro of SU11657, a new compound FLT3 kinase inhibitors, we analyzed human cell lines from AML patients (MV4-11 and HL60) and blast from patients AML using a wide range of concentrations (1nM-10μM) of this novel agent. In HL-60, FLT3-wt cell line, used as negative control does not show relevant effect after treatment with SU11657. Instead, in MV4-11, FLT3-ITD cell line, we observed a decrease dose-dependent in cell viability after treatment with SU11657. The effects of this compound on cell cycle progression show an accumulation of G1/S phase and an induction of apoptosis at 1-10nM concentration after 24h of treatment. First we observed a dephosphorylation of FLT3 on Tyr591 in whole cell extracts from MV4-11 cells after treatment with SU11657 100nM. We also demonstrated a hypophosphorylation of AKT on Ser473 and a consequently dephosphorylation of BAD on Ser136 at nanomolar concentration. We observed a dephosphorylation of STAT-5 to 100nM of SU11657 at 24h.
We evaluated the effects of this new compound in AML primary progenitors that showed FLT3-ITD, FLt3-TKD and FLT3-wt. In the patients with mutation ITD and TKD was evident a modification of cell cycle progression with a decrease in G2/M phase and an increase of subdiploid peak. The effect of SU11657 in patients FLT3-wt was not relevant.
Our study thus showed a potential therapeutic usefulness of the drug in treatment of AML. Study of signal transductions and gene profile expression will contribute to further understanding of the drug mechanisms.
Imaging of Peripheral Blood Mononuclear Cells (PBMC) Trafficking in Patients with Non-Hodgkin’s Lymphoma (NHL) Prior to and Following Therapy.
Authors: Weiner George , Wiseman Gregory , Witzig Thomas , Link Brian , Wooldridge James , Ponto James , Juweid Malik 
Published: April 2020

Abstract
Antibody-dependent cellular cytotoxicity (ADCC) mediated by PBMC’s is believed to play an important role in the anti-tumor effects of rituximab in patients (pts) with NHL but little is known about the trafficking of PBMC’s into the involved nodes. The primary objective of this pilot study is to demonstrate the feasibility of imaging PBMC trafficking in pts with NHL prior to and following various forms of therapy, including rituximab. To date, autologous PBMC’s were harvested in 9 pts (7M, 2F, age 49–83 y) using 100 ml of peripheral blood and Ficoll-Paque density gradient for cell separation and then radiolabeled with 400–450 mCi of 111In-oxine. Total number of leukocytes in the final product ranged from 1.5–12x107 (mean=5.5x107) with %PBMC’s ranging from 40–98 (mean= 75). The radiolabeling efficiency ranged from 53 to 87% (mean = 71) Viability of PBMC’s by trypan blue exclusion was 99–100%. Eight patients were imaged at baseline and six of these also following 2 cycles of rituximab (n=5) or one cycle of R-CHOP (n=1). Imaging was performed at approximately 4, 24, and 48 h after injection of 165–300 mCi of 111In-PBMCs. No significant adverse events were observed. All 8 pts showed the expected biodistribution of PBMC’s in normal organs, such as lung (primarily at 4h) and liver and spleen (at all timepoints). There was no evidence of PBMC localization in normal lymph nodes prior to or posttherapy. In contrast, PBMC localization was seen prior to therapy in ≥ 1 known tumor lesion per pt in all 8 pts and in the 6 pts imaged posttherapy, similar or greater PBMC tumor accumulation was observed. In conclusion, our initial results demonstrate the feasibility of imaging PBMC trafficking in pts with NHL prior to and following therapy with promising initial imaging findings. Our studies could eventually help better understand the role that PBMC play in the anti-tumor effect of rituximab and prove useful in the development of combination treatments such as the use of moAbs and cytokines.
Clinical Significance of Serum Th1-, Th2- and Regulatory T Cells-Associated Cytokines in Adult T-Cell Leukemia/Lymphoma: High Interleukin-5 and -10 Levels Are Significant Unfavorable Prognostic Factors.
Authors: Inagaki Atsushi , Ishida Takashi , Ishii Toshihiko , Komatsu Hirokazu , Iida Shinsuke , Yonekura Kentaro , Takeuchi Shogo , Takatsuka Yoshifusa , Utsunomiya Atae , Ueda Ryuzo 
Published: April 2020

Abstract
Purpose & Experimental Design: Patients with adult T-cell leukemia/lymphoma (ATLL) are in a severely immunocompromised state, leading to frequent and severe infectious complications. Therefore, it is assumed that ATLL cells either express particular cytokines or induce their expression in host immune cells, disrupting the balanced production of cytokines and causing the host’s immune system to break down. We examined the levels of serum cytokines including T helper type 1- (Th1-)associated cytokines (IFN-γ, TNF-α, and IL-2), Th2-associated cytokines (IL-4, -5, and -6), and regulatory T cell-associated cytokines (IL-10 and TGF-β1) in 94 ATLL patients, 39 asymptomatic human T-cell lymphotropic virus type-1 (HTLV-1) carriers, and 50 healthy adult volunteers, in order to clarify whether elevated levels of particular cytokines are associated with the prognosis of ATLL patients.
Results: On multivariate analysis, high IL-5 and IL-10 levels were independent and significant unfavorable prognostic factors among the ATLL patients. The IL-10 level significantly increased with disease progression at each step from asymptomatic HTLV-1 carrier to ATLL of the indolent variant (chronic and smoldering subtypes) to ATLL of the aggressive variant (acute and lymphoma subtypes). Furthermore, high IL-10 was significantly associated with high LDH, indicating that the IL-10 level reflects the tumor burden. The IL-5 level was not associated with disease progression nor LDH. Among ATLL patients with the aggressive variant, high IL-5, but not high IL-10, was an independent and significant unfavorable prognostic factor on multivariate analysis.
Conclusions: The present study provide novel insights into the immunocompromised state of ATLL patients, and show that measurement of serum IL-5 and IL-10 levels is useful for predicting the prognosis and for determining a suitable treatment strategy for ATLL patients.