bluuebunny/crossref_metadata_2025_split huggingface dataset:
Phase I Study of Vaccination with Dendritic Cell Myeloma Fusions.
Authors: Avigan David , Rosenblatt Jacalyn , Vasir Baldev , Wu Zekui , Bissonnette Adam , MacNamara Claire , Uhl Lynne , Lenahan Corrinne , Miller Kenneth , Joyce Robin , Levine James D. , Lowe Karen , Dombagoda Dilani , Richardson Paul , Anderson Kenneth , Munshi Nikhil , Kufe Donald 
Published: October 2019

Abstract
We have developed a cancer vaccine in which patient derived myeloma cells are fused with autologous dendritic cells (DCs). In this way, multiple tumor antigens are presented in the context of the immune stimulating machinery of the DC. We are conducting a phase I trial in which patients with multiple myeloma (MM) undergo serial vaccination with DC/myeloma fusions in conjunction with GM-CSF. Eligibility criteria included ≥20% plasma cells in the bone marrow and measurable paraprotein or light chain. To date, 16 patients have been enrolled and 15 have undergone therapy. Patients underwent leukapheresis collection and DCs were generated from adherent mononuclear cells cultured for 5 days with GM-CSF and IL-4 and matured by exposure to TNFa for 48–72 hours. The mean yield of DCs was 1.23 x108 cells with a mean viability of 88%. Patient derived myeloma cells were isolated from bone marrow aspirates. The mean yield of myeloma cells was 92.5 x 106 cells with a mean viability of 90%. Fusion cells were generated by coculture of DCs with myeloma cells in the presence of 50% polyethylene glycol. Fusion cells were quantified by determining the percentage of cells that co-expressed unique DC and myeloma antigens. Mean fusion efficiency and viability was 40% and 84%, respectively. As a measure of immunologic potency, DC/MM fusions stimulated allogeneic T cell proliferation, with mean stimulation indexes of 48, 36, and 9 for the DC, fusion and myeloma cell populations, respectively. To date, 15 patients have completed vaccination at a dose of 1–5x106 fusion cells. Adverse events potentially related to vaccine included injection site reactions, edema, rash, fever (infection), chills, fatigue, muscle aches, pruritis, and diarrhea. One patient with a history of prior deep venous thrombosis (DVT) developed a DVT and pulmonary embolus of uncertain relation to the vaccine. To date, 6/9 evaluable patients have demonstrated immunologic response, defined by at least 2 fold increase in IFNγ expression by CD4 and/or CD8 T cells in response to ex vivo exposure to autologous tumor lysate. Vaccination was associated with an increase in circulating tumor specific T cells as evidenced by an increase in CD8+ cells binding the MUC1 tetramer. The impact of vaccination on circulating levels of regulatory T cells is being examined. Humoral immune responses are being assessed by SERAX analysis. Three patients have ongoing stable disease at 3.5, 6, and 19 months following their initial vaccination. Six patients demonstrated disease stability with subsequent progression ranging from 9 weeks to 8 months following their first vaccine. Vaccination with DC/MM fusions has been well tolerated, associated with immunologic response, and disease stabilization in a majority of patients with multiple myeloma.
Effects of Celecoxib on Human Acute Promyelocytic Leukemia Cell Line and Its Mechanism.
Authors: Xu Yun , Huang He , Zhao Yanmin , Zeng Fenfang , Zhou Qian 
Published: October 2019

Abstract
Acute promyelocytic leukemia (APL) is a special subtype of acute myelogenous leukemia (AML) which is characterized for a specific translocation between chromosome 15 and 17 [t(15;17)] and the expression of PML/RARα fusion gene. Celecoxib, one of the specific inhibitors of cyclooxygenase-2 (COX-2), has been reported to induce anti-neoplastic activity on many solid human tumor cell lines in recent years. In our study, ATRA resistant APL cell line MR2 cells were used to investigate the effects of celecoxib on hematological malignancy. MR2 cells were treated with celecoxib at different concentration (0, 20, 40, 80, 120 and 160μmol/L). The proliferation of MR2 cells was observed by MTT assay and apoptosis was detected by DNA fragmentation analysis and flow cytometry using Annexin V-FITC/PI staining. Western blot was used to detect the change of caspase-8, -9, -3 and PARP in MR2 cells. The expression of mRNA of fusion gene PML/RARα, COX-2 and survivin, bcl-2/bax, CIAP1 and CIAP2 was assessed by reverse transcription polymerase chain reaction (RT-PCR). Cell cycle analyzed by flow cytometry with PI staining and western blot was used to detect the expression of cell-cycle-regulating proteins. The telomerase activity of MR2 cells was analyzed by PCR-ELISA. The expression of hTERT mRNA and c-myc mRNA was assessed by RT-PCR. MR2 cells viability in presence of celecoxib decreased markedly in a dose- and time- dependent manner. After treated with celecoxib (20-160μmol/L) for 12-48h, the proliferation of MR2 cells were inhibited significantly, in comparison with the control group (P<0.01). 50% growth inhibition (IC50) at 24h and 48h was 80.93μmol/L and 71.72μmol/L, respectively. A DNA ladder pattern of internucleosomal fragmentation was observed. The translocation of phosphatidylserine at the outer surface of the cell plasma membrane could be induced by celecoxib and its level increased following the augmentation of the drug concentration. MR2 cells exposure to 40-160μmol/L celecoxib for 24h caused 9.59%, 24.00%, 36.10% apoptotic cells, which was more than that of the untreated group 2.84% (P<0.01). The expression of survivin mRNA decreased dramatically, while no significant change with PML/RARα and COX-2. Treatment with celecoxib for 24h resulted in the activation of caspase-3 and caspase-9, cleavage of PARP. 40-160μmol/L celecoxib led to cell cycle arrest in G1/S phase, and CyclinD1 and CyclinE decreased, accompanied with up-regulation of P21waf/cip1, P27KIP, P16INK4a. celecoxib could inhibit the telomerase activity of APL cell line, and the inhibition was dose- and time- dependent. The expression of hTERT mRNA and c-myc mRNA were down-regulated by celecoxib in dose- dependent manner. These results indicated that celecoxib could inhibit MR2 cells proliferation by inducing apoptosis, cell cycle arrest and suppression of telomerase activity.
Exclusion from Acute Myeloid Leukemia (AML) Clinical Trials of Patients Unlikely To Respond.
Authors: Tsimberidou Apostolia-Maria , Kantarjian Hagop M. , Faderl Stefan , Freireich Emil J. , Estey Elihu 
Published: October 2019

Introduction: Patients with newly diagnosed AML are increasingly being enrolled on industry-sponsored clinical trials. These trials routinely exclude patients with Zubrod performance status (PS) >2, bilirubin >1.9 mg/dL, or creatinine >1.9 mg/dL; such patients are well-known to be relatively unlikely to respond (UTR). Here we quantify the effect of this practice on inclusion of UTR patients in clinical trials.
Patients and Methods: Overall, 2323 adults with newly-diagnosed AML (no acute promyelocytic leukemia) were registered on clinical trials (industry-sponsored and other) from 1991–2006. Considering patients as UTR if they fulfilled any of the criteria noted above, we examined rates of enrollment of UTR patients over time:
Results: Thus, while more patients are being enrolled on clinical trials, fewer UTR patients are being enrolled particularly over the last 5 years (p = .04) with a concomitant decrease in the proportion of such patients enrolled (p<.0001 over the 16 years in question). Findings are qualitatively the same in older patients.
Conclusion: Newer clinical trials may be less applicable to UTR patients than previous trials. There is a need for clinical trials specific to the UTR population, the group most in need of novel therapies.
Rates of enrollement of unlikely to respond patients on clinical trials for newly diangosed AML Year No. of pts. treated on clinical trial (pts. age≥60 yrs) UTR pts. enrolled (pts. age≥60 yrs) % of UTR pts. enrolled (pts. age ≥60 yrs) 1991 101 (50) 17 (13) 17 (26) 1992 100 (48) 20 (14) 20 (29) 1993 96 (46) 18 (9) 19 (20) 1994 109 (41) 23 (13) 21 (32) 1995 102 (48) 20 (10) 20 (21) 1996 104 (63) 14 (10) 13 (16) 1997 145 (81) 30 (20) 21 (25) 1998 124 (63) 14 (8) 11 (13) 1999 164 (86) 26 (16) 16 (19) 2000 169 (96) 25 (19) 15 (20) 2001 187 (98) 30 (19) 16 (19) 2002 162 (91) 17 (11) 10 (12) 2003 195 (111) 4 (2) 2 (2) 2004 184 (109) 17 (11) 9 (10) 2005 185 (117) 15 (7) 8 (6) 2006 196 (109) 17 (12) 9 (11) Total 2323 (1257) 307 (194) 13 (15)
Efficacy of Nilotinib (AMN107) in Patients (Pts) with Newly Diagnosed, Previously Untreated Philadelphia Chromosome (Ph)-Positive Chronic Myelogenous Leukemia in Early Chronic Phase (CML-CP).
Authors: Cortes Jorge , O’Brien Susan , Jabbour Elias , Ferrajoli Alexandra , Borthakur Gautam , Burger Jan , Wierda William , Garcia-Manero Guillermo , Thomas Michelle , Letvak Laurie , Kantarjian Hagop 
Published: October 2019

Abstract
Background: Nilotinib is a novel, oral tyrosine kinase inhibitor with increased selectivity against Bcr-Abl that is approximately 30-fold more potent than imatinib. High response rates have been reported with nilotinib therapy in CML after imatinib failure.
Methods: We evaluated the efficacy of nilotinib as first line therapy in pts with newly diagnosed Ph-positive CML-CP. The primary objective was to estimate the proportion of pts attaining major molecular response (BCR-ABL/ABL ratio ≤0.05% in our lab) at 12 months (mo).
Results: Thirty-two pts have been treated with nilotinib 400 mg orally twice daily for a median of 5 months (mo) (range, 1 to 31 mo). The median age was 47 years (yrs) (range, 24–73 yrs). The Sokal risk at pretreatment was low in 21 (70%) pts, intermediate in 6 (20%), and high in 3 (10%). The rate of complete cytogenetic response [CCyR] (Ph 0%) at 3, 6 and 12 mo compares favorably to those observed in historical controls treated with imatinib 400 mg or 800 mg daily:
The median QPCR with nilotinib at 3, 6, and 12 months were, respectively, 0.52% (range, 0.0–29.5%), 0.03% (range, 0.0–9.13%), and 0.09% (range, 0.0–16.21%). At 3 mo follow-up, major molecular response (MMR; BCR-ABL/ABL ratio ≤0.05%) was observed in 3/22 patients (14%), 7/13 (54%) at 6 mo, and 5/11 (45%) at 12 mo. 12-mo rates of MMR for the historical imatinib groups treated at 400 mg and 800 mg were 24% and 47%, respectively (p=0.02). None of the molecular responses has been lost while on therapy. Grade 3–4 neutropenia was observed in 2 (7%) pts, and thrombocytopenia in 1 (3%). Other grade 3–4 adverse events included elevation of lipase (n=3, 9%), or bilirubin (n=2; 6%), and amylase elevation, back pain, and infection (1 each). Twelve pts had transient treatment interruptions (median 11 days), most frequently due to pain (n=3; musculoskeletal 2, abdominal 1), lipase elevation (n=2). Seven patients had their dose reduced to 400 mg daily, 2 to 200 mg twice daily, and 1 to 200 mg daily due to extramedullary toxicity. Three patients decided to change therapy after 4, 6 and 8 months; 2 switched to imatinib and 1 received SCT.
Conclusion: Nilotinib 400 mg orally twice daily suggest significant efficacy manifested by complete cytogenetic responses in nearly all patients as early as 3 months after the start of therapy with a favorable toxicity profile.
Percent with CCyR (No. evaluable) Months on Therapy Nilotinib Imatinib 400 mg Imatinib 800 mg P value 3mo 95 (22) 37 (49) 62 (202) < 0.0001 6mo 100 (13) 54 (48) 82 (199) <0.0001 12mo 100 (11) 65 (48) 86 (197) 0.0007
Inhibitory Effects of Homoharringtonine on Leukemic Stem Cells and BCR-ABL Induced Chronic Myeloid Leukemia and Acute Lymphoblastic Leukemia in Mice.
Authors: Chen Yaoyu , Hu Yiguo , Michaels Shawnya , Brown Dennis , Li Shaoguang 
Published: October 2019

Abstract
The Abl tyrosine kinase inhibitors (TKIs) imatinib mesylate (IM) and dasatinib, targeting BCR-ABL for the treatment of Philadelphia-positive (Ph+) leukemia including chronic myeloid leukemia (CML) and B-cell acute lymphoblastic leukemia (B-ALL), have produced impressive results in terms of therapeutic outcome and safety for patients. However, clinical resistance to these TKIs likely at the level of leukemic stem cell negates curative results in Ph+ leukemia. At present, an anti-stem cell strategy has not been developed for treating these leukemia patients. Homoharringtonine (HHT) (omacetaxine mepesuccinate - USAN/INN designation) has shown significant clinical activity in CML in combination with IM or alone for patients failing IM. However, little is known about whether HHT has an inhibitory effect on leukemic stem cells. The purpose of this study is to determine whether HHT inhibits BCR-ABL-expressing leukemic stem cells (Lin-c-Kit+Sca-1+) that we identified previously (Hu et al. Proc Natl Acad Sci USA 103(45):16870–16875, 2007) and to evaluate therapeutic effects of HHT on CML and B-ALL in mice. We find that in our in vitro stem cell assay, greater than 90% of leukemic stem cells were killed after being treating with HHT (12.5, 25, and 50 nM) for 6 days, and in contrast, greater than 75% or 92% of leukemic stem cells survived the treatment with dasatinib (100 nM) or imatinib (2 mM). We next treated CML mice with HHT (0.5 mg/kg, i.p., once a day). 4 days after the treatment, FACS analysis detected only 2% GFP+Gr–1+ myeloid leukemia cells in peripheral blood of HHT -treated CML mice and in contrast, 41% GFP+Gr–1+ myeloid leukemia cells in placebo-treated mice. We also treated mice with BCR-ABL induced B-ALL with HHT, and found that only 0.78% GFP+B220+ lymphoid leukemia cells were detected in peripheral blood compared to 34% GFP+B220+ lymphoid leukemia cells in placebo-treated mice. Furthermore, HHT significantly inhibited in vitro proliferation of K562 and B-lymphoid leukemic cells isolated from mice with B-ALL induced by BCR-ABL wild type and BCR-ABL-T315I resistant to both imatinib and dasatinib. In sum, HHT has an inhibitory activity against CML stem cells, and is highly effective in treating CML and B-ALL induced by BCR-ABL in mice.
Molecular Basis Explanation of Imatinib Resistance of Bcr-Abl Due to T315I and P-Loop Mutations from Molecular Dynamics Simulations.
Authors: Lee Tai-Sung , Potts Steven , Kantarjian Hagop , Cortes Jorge , Giles Francis , Albitar Maher 
Published: October 2019

Abstract
Molecular dynamics (MD) simulations on the complex of imatinib with the wild-type, T315I, and other 10 P-loop mutants of the tyrosine kinase Bcr-Abl have been performed to study the imatinib resistance mechanism at the atomic level. MD simulations show that large scale computational simulations could offer insight information that a static structure or simple homology modeling methods cannot provide for studying the Bcr-Abl imatinib resistance problem, especially in the case of conformational changes due to remote mutations. By utilizing the Molecular Mechanics/Poisson-Boltzmann surface area (MM-PBSA) techniques and analyzing the interactions between imatinib and individual residues, imatinib resistance mechanisms not previously thought have been revealed. Non-directly contacted P-loop mutations either unfavorably change the direct electrostatic interactions with imatinib, or cause the conformational changes influencing the contact energies between imatinib and other non-P-loop residues. We demonstrate that imatinib resistance of T315I mainly comes from the breakdown of the interactions between imatinib and E286 and M290, contradictory to previously suggested that the missing hydrogen bonding is the main contribution. We also demonstrate that except for the mutations of the direct contact residues, such as L248 and Y253, the unfavorable electrostatic interaction between P-loop and imatinib is the main reason for resistance for the P-loop mutations. Furthermore, in Y255H, protonation of the histidin is essential for rendering this mutation resistant to Gleevec. Our results demonstrate that MD is a powerful way to verify and predict clinical response or resistance to imatinib and other potential drugs.
Glycoprotein Ibα Forms Catch Bonds with von Willebrand Factor A1 Domain but Not with Mutant A1 Domains Exhibiting Properties of Type 2B von Willebrand Disease.
Authors: Wu Tao , Yago Tadayuki , Yang Jun , Miner Jonathan , Coburn Leslie , Lopez Jose A. , Cruz Miguel A. , McIntire Larry V. , McEver Rodger P. , Zhu Cheng 
Published: October 2019

Abstract
Interactions between glycoprotein Ibα (GPIbα) and von Willebrand factor (VWF) initiate platelet adhesion to injured vascular surfaces, which is enhanced by arterial blood flow. The flow requirement for adhesion is reduced (i.e., gain-of-function) by single-residue substitutions of the VWF-A1 domain, e.g., R543Q that occurs in some patients with type 2B von Willebrand disease (VWD) and R687E that was designed to exhibit type 2B VWD properties. Yet the mechanisms for flow-enhanced adhesion through GPIbα-VWF interactions are not understood. By probing single bonds with atomic force microscopy, we showed that lifetimes of GPIbα/VWF-A1 bonds first increased (catch) and then decreased (slip) with increasing force applied to the bond. Remarkably, the catch bond aspect of the GPIbα/VWF-A1 bonds was eliminated by the R543Q and R687E mutations, which exhibited slip bonds only with prolonged lifetimes at low forces. Flow chamber experiments showed that catch-slip transitional bonds governed flow-enhanced rolling of platelets and GPIbα-coated microspheres on wild-type A1, such that rolling velocities first decreased and then increased with increasing flow. By contrast, slip bonds governed rolling velocities on R543Q and R687E A1 mutants, which increased monotonically with increasing flow. We changed fluid viscosity by adding Ficoll to the medium, tether force by using microspheres of different radii, and platelet deformability by using a chemical fixative. The rolling velocity vs. flow curves aligned with tether force but not with transport parameters and were minimally affected by fixation, which respectively rule out transport-enhanced GPIbα/VWF-A1 bond formation and force-induced enlargement of platelet-surface contact area as the causes for flow-enhanced rolling. Flowing platelets agglutinated with microspheres bearing R543Q and R687E A1 mutants but not with those bearing wild-type A1, suggesting that GPIbα/VWF-A1 catch bonds prevent agglutination of circulating platelets via binding to VWF multimers and that platelet agglutination in patients with type 2B VWD may be explained by the prolonged lifetimes at low forces resulting from elimination of catch bonds in the interactions of GPIbα with VWF-A1 mutants.
Glyoxalase-I Is a Potential Therapeutic Target for Chronic Myelogenous Leukemia Cells That Have Acquired Drug Resistance through Adaptation to Hypoxia.
Authors: Takeuchi Miki , Kuroda Junya , Kimura Shinya , Ashihara Eishi , Umezawa Kazuo , Yasui Eiko , Kawatani Makoto , Osada Hiroyuki , Hodohara Keiko , Fujiyama Yoshihide , Maekawa Taira 
Published: October 2019

Abstract
Growth under hypoxic conditions can lead to resistance to a variety of cell death stimuli in cancer cells. Bone marrow (BM) has less vasculature than other organs, and the rapid growth of leukemic cells in BM frequently outpaces the oxygen supply. Thus, adaptation to hypoxia may be critical to the survival and proliferation of leukemic cells. However, the role of hypoxia in the treatment of leukemia has received little attention. We hypothesized that leukemic cells acquire drug resistance by adapting to hypoxia in the BM, and that identifying the underlying molecular mechanisms for this adaptation would reveal new approaches to the development of new therapeutic targets in drug- resistant leukemia cells. To test this hypothesis, we cultured five chronic myelogenous leukemia (CML)-derived cell lines (BV173, K562, KCL22, KT-1, and MYL) continuously in 1.0% oxygen. Among them, hypoxia resistant (HR) subclones of K562 and KCL22 cells (K562/HR and KCL22/HR) were selected under 1.0% oxygen conditions, and these cells were able to grow for several months and to be suspended in RPMI1640 plus 10% FCS without any additional nutrients, although their growth rate was slower than parental cells in 20% oxygen. Both K562/HR and KCL22/HR cells produced less ATP (78% and 92%, respectively), perhaps due to impairment of the oxygen-dependent energy production pathway, and HR cells showed higher expression and enhanced kinetics of glyoxalase-I (Glo-I), a detoxification kinase of methylglyoxal (a cytotoxic byproduct of glycolysis), compared to parental cells. Glo-I has previously been implicated in drug resistance in cancer, including leukemia. Autophosphorylation of Bcr-Abl was reduced in HR cell lines, while the phosphorylation status of the downstream signalling molecules Akt and Stat5 was unchanged, which suggests that there are alternative, hypoxia-specific activators of these pathways. K562/HR and KCL22/HR cells were less sensitive to chemotherapeutic agents such as the Bcr-Abl tyrosine kinase (TK) inhibitors imatinib, dasatinib, nilotinib, and INNO-406, and the alkylating agents daunorucbicin and busulfan. In contrast, K562/HR and KCL22/HR cells were more sensitive to cell death induced by a Glo-I inhibitor, COTC, compared to parental cells. These results indicated that Glo-I plays an important role in survival under hypoxic conditions, and that Glo- I may be an alternative molecular target in CML cells that have acquired resistance to conventional chemotherapeutics through adaptating to hypoxia.
Combination of Imatinib with Cisplatin and Nutlin-3: Functional and Molecular Effects on Bcr-Abl Positive Cells.
Authors: Skorta Ioanna , van der Kuip Heiko , Henkes Martin , Oren Moshe , Aulitzky Walter E. 
Published: October 2019

Abstract
Imatinib is highly effective in inducing remissions in chronic phase CML. However, complete eradication of the malignant clone by Imatinib monotherapy is rare. This prompted us to explore the efficacy of combination of Imatinib with the DNA damaging agent Cisplatin and with Nutlin-3, a compound which induces p53 accumulation by interfering with its binding to Mdm2. We used a Bcr-Abl positive cell line characterized by the loss of Imatinib sensitivity in the presence of optimal growth factor concentrations. Whereas combined treatment of Imatinib with either Cisplatin or Nutlin-3 in low and nontoxic doses induced ∼15–20% cell death, simultaneous treatment of all three compounds was highly effective (∼50% cell death) even in the presence of optimal growth conditions. Importantly, comparable effects were also seen in CFU assays performed with primary CD34 positive cells from 5 CML patients. To examine the molecular mechanisms of these effects we analyzed p53 dependent cellular response to Cisplatin in our cell line model in the presence or absence of Imatinib and/or Nutlin-3. The active Bcr-Abl kinase caused superinduction of p53 protein after DNA damage. We found both an upregulated p53 accumulation and enhanced p21 and Mdm2 RNA and protein levels upon Cisplatin treatment. This phenomenon could be reversed both by siRNA-mediated inhibition of Bcr-Abl expression and by Imatinib-induced inhibition of Bcr-Abl kinase activity: despite of optimal growth conditions inhibition of Bcr-Abl caused a significant reduction of p53 expression and activation. In the presence of Nutlin-3, p53 expression was significantly upregulated upon Cisplatin treatment both with and without Imatinib. However, Nutlin-3 was not capable to restore the Imatinib-induced blockade of the transcriptional activity of p53 on mdm2. The reduced p53 activation observed in Bcr-Abl positive cells treated with Imatinib was paralleled by a shift from cell cycle arrest to cell death both in the presence and absence of Nutlin selectively in Bcr-Abl positive cells rendering them hypersensitive to Cisplatin. In summary, combined treatment of Imatinib with low concentrations of a DNA damaging agent and a p53 activator is highly effective in vitro. Such combined treatment may prove to be clinically relevant for complete eradication of the malignant clone in CML, provided that conditions are found where it does not affect adversely normal hematopoietic cells in vivo.