bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of GSK3 in Multiple Myeloma Cell Growth and Survival.
Authors: Piazza Francesco A. , Gurrieri Carmela , Chioetto Gino , Colpo Anna , Bonanni Luca , Montini Barbara , Tubi Laura Quotti , Gnoato Marianna , Carraro Samuela , Cabrelle Anna , Trentin Livio , Zambello Renato , Semenzato Gianpietro 
Published: April 2020

Abstract
The serine-threonine GSK3 displays a crucial role in different cancer-pathogenetic pathways, including the PI3K/AKT, Wnt β-catenin and NF-κB signaling cascades, either promoting or counteracting cell survival. The aim of this study was to investigate the role of GSK3 in multiple myeloma (MM) cell growth. GSK3α and β total and phosphorylated protein levels were found differentially expressed in malignant plasma cells as compared to normal resting B-lymphocytes and to normal in vitro generated plasmablasts. Intriguingly, in freshly isolated malignant plasmacells from patients, most of GSK3 was found colocalized with Wnt receptor LRP6 and casein kinase I next to the cell membrane as compared to normal plasmacells or B-cells from other malignancies, wher it was distributed in the cytosol and in the nucleus, thus suggesting a peculiar role of this kinase in these cells. Upon stimulation of MM cells with IL-6 and IGF-I, GSK3 enzymatic activity was hampered, while stimulation with TNFα did not affect GSK3 function nor the early events in NF-κB activation. Basal and IL-6 and IGF-I driven proliferation of MM cells was slightly impaired by GSK3 blockade. Interestingly, GSK3β−/− mouse embryo fibroblasts (MEFs) proliferated slightly slower as compared to GSK3β+/+ cells; however, GSK3α inhibition and IL-6 and IGF-I stimulation, resulted in much higher proliferation of GSK3β −/− cells. Intriguingly, GSK3 inhibition with specific compounds (SB216763 and SB415286) caused a significant rescue from cell death of growth factor-deprived MM cells while resulted in reduced cell proliferation and apoptosis of MM cells grown with serum or growth factors. When GSK3 inhibitors were added to MM cell cultured with bone marrow stromal cells (BMSC), MM cells survival increased and NF-κB and β-catenin-mediated gene transcription (of IAPs and cyclinD1, respectively) was deregulated. GSK3 activity inhibition did not modify the rate of proteasome inhibitor-induced cell death in co-colture experiments with BMSC, suggesting that the sensitivity to bortezomib-induced MM cell apoptosis is independent on GSK3. Altogether, our data indicate thatGSK3 localizes on the cell membrane in primary MM cells;GSK3 is differently regulated downstream from growth factors or TNFα-induced signaling pathways in MM cells;a peculiar role of GSK3 in malignant MM cells as compared to normal MEFs with regard to cell proliferation; anda critical role of this kinase in regulating the MM microenvironment.
Pre-Clinical Validation of Polo-Like Kinase 1 as a Therapeutic Target in Multiple Myeloma with the Selective Inhibitor BI2536.
Authors: Sidhu Roger , Steffler Tara , Hipperson Gail , Jung Michelle , Fung Joyce , Pilarski Linda M. , Belch Andrew R. , Reiman Tony 
Published: April 2020

Abstract
BACKGROUND: The centrosome is the cellular organelle that nucleates the mitotic spindle. Polo-like kinase 1 (Plk1), a centrosome-associated serine/threonine kinase, serves as a key regulator of multiple steps in mitosis. Plk1 is overexpressed in a broad spectrum of tumor types, and its expression often correlates with poor patient prognosis. We have previously observed centrosome amplification in myeloma, and have validated other components of the myeloma centrosome and mitotic apparatus as therapeutic targets. The present study explored the expression of Plk1 in myeloma and the effect of BI2536, a potent and selective inhibitor of Plk1, on myeloma cells in the pre-clinical setting.
METHODS: Plk1 expression was assayed in a panel of myeloma cell lines (MMCL) and AutoMACS-purified CD138+ patient bone marrow plasma cells (BMPC). The anti-myeloma effects of BI2536, alone or in combination with conventional agents, were assayed on myeloma cells with proliferation (MTS) and apoptosis (Annexin V/propidium iodide) assays. The phenotype of treated cells was examined with DNA content analysis and immunofluorescence microscopy. The efficacy of BI2536 monotherapy was evaluated in NOD/SCID mice bearing RPMI8226 xenografts.
RESULTS: Plk1 is ubiquitously expressed in myeloma to varying degrees in both MMCL and BMPC. BI2536 inhibited the proliferation of MMCL (RPMI 8226, U266, LP-1 and KMS-11) and patient BMPC at nanomolar concentrations. The addition of BI2536 was able to overcome resistance to dexamethasone. Bortezomib in combination with BI2536 had significantly increased anti-myeloma effects compared to the use of either agent alone. BI2536-treated MMCL accumulated 4N DNA content prior to undergoing apoptosis. The phenotype of BI2536-treated cells is consistent with inhibition of Plk1, showing prometaphase arrest and monopolar mitotic spindles in a dose-dependent fashion. BI2536 induces regression of human myeloma xenografts in NOD/SCID mice. Taken together, BI2536 is a promising new agent for the treatment of multiple myeloma. This work provides further evidence that Plk1 and the amplified myeloma centrosome are targets for therapy.
Cucurbitacin I (JSI-124) Has Potent Anti-Myeloma Effects Independent of Its Inhibition of JAK2-Induced STAT3 Activation.
Authors: Ma Huihui , Ziegler Judith , Feng Ren-Tien , Lentzsch Suzanne , Mapara Markus 
Published: April 2020

Abstract
Multiple myeloma (MM) is a plasma cell proliferative disorder that results in considerable morbidity and mortality. JSI-124 is a plant natural product identified previously as Cucurbitacin I, isolated from various plant families such as the Cucurbitaceae and Cruciferae and has been recently described as a specific inhibitor of Janus Kinase-2 (JAK-2)/signal transducer and activator of transcription-3 (STAT3). Based on the critical role of the IL-6/STAT3 pathway in MM we studied the effects of JSI-124 on different MM cells in vitro. Different human myeloma cell lines including MM1.S, IM9, OPM-2, RPMI-8226, ARH77 in addition to the murine 5TGM myeloma cell lines were incubated with increasing concentrations of JSI-124. The impact of JSI-124 on cell proliferation, cell cycle and induction of apoptosis was studied using [3H]-Thymidine incorporation, cell counting, flow cytometry and Annexin/PI staining and caspase 3, 8 and 9 activation. JSI-124 was able to inhibit proliferation and induce apoptosis in several MM cell lines, including MM1.S, IM9, OPM-2, RPMI-8226, ARH77, U266 and 5TGM in a dose- and time-dependent manner. JSI-124 lead to activation of caspase 3, 8 and 9 indicating involvement of both extrinsic and intrinsic apoptotic pathways. JSI-124-mediated induction of apoptosis was independent of JAK2/STAT3 inhibition as MM1.S and 5TGM cells, which lack constitutive STAT3 (Tyr705) activation were equally sensitive to JSI-124 compared to U266 cells which show a constitutive STAT3 Tyr705 phosphorylation. However, JSI-124 treatment was able to abrogate IL-6 and bone marrow stroma (BMSC)-induced STAT3 (Tyr705) activation in MM1.S cells. In addition we were able to observe JSI-124 dependent inhibition of constitutive STAT3 (Ser727) activation in MM cell lines. To further delineate the mechanism underlying its anti-myeloma effects we studied the impact of JSI-124 treatment on NF-κB, MAPK and PI3K pathways. Indeed, JSI-124 treatment resulted in inhibition of p-p65, p-MEK1,2 and p-Akt underscoring the effect of JSI-124 on STAT3-independent signaling. Our results indicate that JSI-124 is a powerful direct inhibitor of myeloma cells blocking constitutive and IL-6/BMSC-dependent STAT3 activation in addition to STAT3 independent signaling pathways. JSI-124 might therefore serve as a potent novel anti-myeloma agent targeting both myeloma cells and its bone marrow microenvironment. Further studies are warranted to evaluate the in vivo efficacy of JSI-124 and identify the STAT3 independent pathways contributing to myeloma cell growth and induction of apoptosis.
Concordance of Assays Designed for the Quantitation of JAK2 1849G&gt;T (V617F): A Multi-Centre Study.
Authors: Lippert Eric , Girodon François , Hammond Emma , Carillo Serge , Richard Céline , Fehse Boris , Hermans Mirjam , James Ian , Jelinek Jaroslav , Marzac Christophe , Migeon Marina , Pietra Daniela , Prchal Josef T. , Reading N. Scott , Sobas Marta , Ugo Valérie , Skoda Radek C. , Hermouet Sylvie 
Published: April 2020

Abstract
Studies of myeloproliferative disorders (MPDs) aiming to evaluate the fraction of the clone bearing the JAK2 1849T mutation occasionally report discordant findings. One reason could be different sensitivity and accuracy of the various assays designed for the detection and quantitation of JAK2 1849G&gt;T. We studied the concordance of 10 published JAK2 1849G&gt;T assays. 29 samples of genomic DNA were distributed to 14 laboratories in France, USA, Australia, Germany, Holland, Italy and Switzerland for blinded assessment of JAK2 1849T levels. DNA was extracted from granulocytes of patients diagnosed with MPD, eosinophilia or secondary polycythemia. The 10 assays tested included 5 TaqMan assays with specificity based on primers (4) or competing probes (1) and allele-specific PCR (AS-PCR) (3), pyrosequencing (1) and FRET/melting curve (1) assays. Standards used for calibration were dilutions of DNA from plasmids (3 sets), cell lines or patient granulocytes. Results were expressed as %1849T allele/total JAK2 (13 centres) or as %1849T allele/control gene (1 centre). One centre had one false negative result; there were no false positive results. PCR equipment did not significantly affect the quantitation of 1849T: after adaptation of the technique, one centre tested one AS-PCR assay on 2 apparatus, 5 other centres tested one TaqMan assay on 3 apparatus; comparable results were obtained in the 6 centres. For 6 assays (10 centres), quantitation in the 26 positive samples, ranging from 1% to 96%, did not differ significantly. Overall variation was 30%; concordance improved with increasing mutational load (18% variation for samples with &gt;8% 1849T). Three TaqMan and 1 AS-PCR assays gave significantly different results, 2 with overall low quantitation. For 3 assays, discordance was explained by an incorrect estimation of 1849T content in the standards. For the 4th discordant assay, expressing results as % 1849T/control gene, values tended to be higher proportional to the consensus. Interestingly, results were consistent with the presence of &gt;2 copies of JAK2 per cell in 4 samples. The study underlined the importance of using defined standards when analysing JAK2 1849T levels. After adaptation to the equipment and with the use of correct standards, all assays gave comparable quantitation of JAK2 1849T, with a sensitivity &lt;1%. Finally, quantitation of a second gene, in order to detect additional copies of JAK2 (&gt;2/cell), should be considered.
JAK2 Mutations at Exon 12 and 14 in Polycythemia Vera and Idiopathic Erythrocytosis: Incidence and Correlation with Clinical Characteristics.
Authors: Besses Carlos , Martínez-Avilés Luz , Alvarez-Larrán Alberto , Cervantes Francisco , Hernández-Boluda Juan Carlos , Colomer Dolors , Florensa Lourdes , Serrano Sergi , Bellosillo Beatriz 
Published: April 2020

Abstract
Introduction. Exon 12 mutations of the JAK2 gene have been described in polycythemia vera (PV) and idiopathic erythrocytosis (IE) patients. These patients display a clinical phenotype different to that observed in V617F-positive PV patients, but no information is available on their clinical outcome.
Patients and methods. Twenty JAK2 V617F-negative PV or IE patients and 86 V617F-positive PV patients were assessed for exon 12 and exon 14 mutations. Analysis of cell lineage mutational status was assessed following cell sorting. Aim. To analyze the presence of JAK2 mutations at exon 12 and 14 in a cohort of V617F-negative PV and IE patients and to correlate them with the patient clinicohematological and evolutive data.
Results. Exon 12 mutations were detected in 4 (20%) V617F-negative patients (K539L, two cases, N542_E543del and H538_K539delinsL, one case each). In 16 patients, no mutations were found in either exon 12 or exon 14. Additional mutations in exon 14 were found in two V617F-positive patients. Two patients with exon 12 mutations developed thrombosis after diagnosis, with the probability of thrombosis-free survival being 75% at 5 years. Another patient with exon 12 mutation developed myelofibrosis at 20 years of PV diagnosis. JAK2 mutations were present in granulocytes, platelets and monocytes, but not in lymphocytes or NK cells.
Conclusions. Patients harboring exon 12 mutations represent a subset of JAK2 V617F-negative PV or IE patients. These patients have initial hematological data different from V617F-positive patients, but they do not differ with regard to thrombosis development and myelofibrotic transformation.
Table 1: JAK2 exon 12 and exon 14 mutations in blood cell subpopulations. Mutation Granulocytes Platelets CD14+ CD3+ CD19+ CD56+ * exon 12; # exon 14; + indicates the presence of the mutation; − indicates no detectable mutation ; ND not determined Patient 1* K539L (AAA →TTA) + + − − − − Patient 2* K539L (AAA →CTA) + + + − − − Patient 3* H538_K539delinsL + + ND − − ND Patient 4* N542_E543del + + + − − − Patient 5# V617F (GTC →TTT) + + − − − ND C618R (TGT →CGT) + + − − − ND Patient 6# V617F (GTC →TTC) + + + − − ND C616C (TGT →TGC) + + + − − ND
mTOR Inhibition for Relapsed or Refractory Hodgkin Lymphoma: Promising Single Agent Activity with Everolimus (RAD001).
Authors: Johnston Patrick B. , Ansell Steven M. , Colgan Joseph P. , Habermann Thomas M. , Inwards David J. , Micallef Ivana N.M. , Porrata Luis F. , Reeder Craig B. , Roy Vivek , LaPlant Betsy R. , Witzig Thomas E. 
Published: April 2020

Abstract
Background: mTOR inhibition has produced responses in mantle cell lymphoma as well as other non-Hodgkin lymphomas. This phase II study tested the oral mTOR inhibitor everolimus (RAD001, Novartis Pharmaceuticals) in simultaneous two-stage phase II lymphoma studies, including one arm involving Hodgkin lymphoma. The goals were to learn the toxicity profile and to assess the anti-tumor response. A total of 17 patients with Hodgkin lymphoma were enrolled in the uncommon arm at completion of enrollment.
Methods: Patients (pts) received 10 mg PO daily for each 28 day cycle (up to 12, with a possible extension in responders) and restaged after 2, 6, and 12 cycles. The primary endpoint is the confirmed response rate, including CR, CRu or PR.
Results: The median age of the 17 pts with Hodgkin lymphoma was 37 yrs (range: 27–68), with a median of 6 (range, 4–14) prior therapies. Fourteen pts (82.4%) had a prior stem cell transplant (SCT). Pts completed a median of 6 (range, 1–13) cycles of therapy. Fifteen of 17 patients were evaluable for response as of this analysis. The overall response rate was 47% (7/15), all partial responses. Ten patients are continuing on study while 6 have gone off due to disease progression and 1 due to other reasons. Common grade 3 adverse events (AEs) include thrombocytopenia (5 pts), anemia (5 pts) and alkaline phosphatase elevation (1 pts). 1 patient was reported to have grade 4 neutropenia.
Conclusions: Oral everolimus has promising activity with acceptable toxicity in Hodgkin lymphoma. These results provide the rationale for additional studies with this novel class of agents and to integrate mTOR inhibitors into salvage treatment regimens for Hodgkin lymphoma.
Vorinostat (Suberoylanilide Hydroxamic Acid) in Relapsed or Refractory Hodgkin Lymphoma: SWOG 0517.
Authors: Kirschbaum Mark H. , Goldman Bryan H. , Zain Jasmine M. , Cook James R. , Rimsza Lisa M. , Forman Stephen J. , Fisher Richard I. 
Published: April 2020

Abstract
Background: Patients with Hodgkin’s Lymphoma with relapse or refractory disease after salvage therapy and transplant face a paucity of active agents. New modalities of treatment are necessary. Vorinostat (SAHA) is an orally administered hydroxamic acid histone deacetylase inhibitor with activity against class I and II deactylases. After initial phase I data (Kelley WK, et al 2005) showed prolonged stable disease in patients with Hodgkin’s, a Phase II study of this agent was launched in patients with relapsed or refractory Hodgkin’s lymphoma.
Methods: Eligible patients had relapsed or refractory Hodgkin’s lymphoma, bidimensionally measurable disease, and performance status 0-2. Patients may have received up to five prior chemotherapy regimens; previous autologous transplant is allowed. Vorinostat was dosed at 200 mg po twice daily for 14 consecutive days on a 21 day cycle. CT scanning was performed after every three cycles, as was marrow biopsy for those with marrow involvement at time of entry into study. This was a 2-stage design with a maximum planned accrual of 35 eligible patients. Objective response was the primary endpoint.
Results: Of 27 patients accrued to the first stage of this study, 25 are eligible (14 males, 11 females). Median age is 41 (19–71) years. Toxicities attributable to drug of grade 3 or higher include fatigue, anorexia, anemia, and thrombocytopenia. One partial response was observed, for a response probability of 4% (95% CI: 0%, 20%). Although there were not adequate responses to open the second stage of accrual, four patients had stable disease lasting at least one year. Of these four, one opted for radiation at 12 months, another progressed at 14 months, whereas two continue on treatment, one with bulky disease who is stable at 12 months, one with symptomatic lung involvement and bulky disease who remains stable at 16 months. One additional patient remains on treatment with stable disease at 9 months.
Conclusions: Although the low response rate led to closure of this trial after the first stage of accrual, we observed that treatment with the oral histone deacetylase inhibitor Vorinostat can lead to protracted stable disease in patients with relapsed/refractory Hodgkin’s Lymphoma. Studies in combination with other molecular agents or chemotherapy may be warranted.
Arsenic Trioxide (As2O3) Mediated Cell Death in Hodgkin Lymphoma (HL) and Non-Hodgkin Lymphoma (NHL): Investigation of Reactive Oxygen Species (ROS), Caspase, and MAP Kinase Signaling Pathways.
Authors: Bhalla Savita , Singh Amareshwar T.K , Prachand Sheila , Haque Qudsia , O’Halloran Thomas V. , Winter Jane N. , Schumacker Paul T. , Platanias Leonidas C. , Gordon Leo I. , Evens Andrew M. 
Published: April 2020

Abstract
The cytotoxic activity of As2O3 in leukemia and myeloma has been shown to be mediated in part by ROS. We determined the mechanism of cytotoxicity of As2O3 in HL and NHL cell lines. Ramos, HF-1, SUDHL4 (NHL lines), and L428 (HL line) cells were cultured with increasing clinically relevant As2O3 concentrations (1.0mm-10mm) at 24-72 hours with and without the oxidizing agent, buthionine sulfoxime (BSO, 100mm), and with and without caspase inhibitors Z-VAD-FMK and Z-IETD-FMK. Apoptosis was measured by Annexin-V/propidium iodide (PI) staining and detected by flow cytometry (FACS). ROS was measured by oxidation of 2′7′ dichlorofluorescein diacetate (H2DCFDA) to Dichlorofluorescein (DCF) and analyzed by FACS. Immunoblots were performed for bcl-2, PARP, cleaved caspases 3, 8, 9, and mitogen activated protein kinase (MAPK) pathways (ERK, JNK, and p38 activation). As2O3 alone at 10mM resulted in dose- and time-dependent apoptosis in all cell lines (HL and NHL) with &gt;75% annexin+/PI+ between 48 and 72 hours. In Ramos and L428 cells, a combination of As2O3 (2 mM) and BSO 100mM resulted in &gt;80% annexinV+/PI+, while either agent alone resulted in &lt;15% apoptosis (p=0.001). A four-fold increase in ROS was seen in all four cell lines with As2O3/BSO, but not with As2O3 alone. Both ROS production and apoptosis induced by As2O3/BSO and As2O3 alone were reversible with the anti-oxidant N-acetylcysteine (NAC) (10mM). Furthermore, As2O3/BSO induced PARP cleavage and caspase-3 activation in all NHL cell lines, but not in L428. In Ramos, Z-VAD-FMK and Z-IETD-FMK blocked As2O3-induced apoptosis (suggesting a caspase-dependent mechanism), but had no effect on As2O3/BSO-induced cell death (caspase-independent). To investigate the signaling pathways involved, we performed western blot analysis for phospho-p38 (p-p38), phospho-JNK and phospho-ERK (p-ERK) in Ramos and L428 cells following As2O3 alone and As2O3/BSO. We found up-regulation of p-p38 in Ramos cells, while in L428, p-ERK was activated following As2O3 and As2O3/BSO. Inhibitors of p-38 in Ramos cells resulted in up-regulation of ERK. In summary, As2O3 induced dose- and time-dependent apoptosis in HL and NHL cell lines was significantly enhanced with the addition of BSO, and inhibited by NAC, suggesting that apoptosis was in part ROS-dependent. In Ramos, As2O3 alone resulted in caspase-dependent apoptosis, while addition of BSO resulted in caspase-independent apoptosis. As2O3 alone and combined with BSO resulted in activation of MAPK pathways in HL and NHL lines. These data provide a basis and a mechanism for As2O3-induced cell death in HL and NHL and have therapeutic implications.
Long Term Outcome of Gastric MALT Lymphoma Patients Treated with Anti-Helicobacter (Antibiotic and Proton-Pump Inhibitor) Regimens.
Authors: Chini Claudio , Pinotti Graziella , Stathis Anastasios , Proserpio Ilaria , Bertoni Francesco , Cavalli Franco , Zucca Emanuele 
Published: April 2020

Abstract
Background: Primary mucosa-associated lymphoid tissue (MALT) lymphoma of the stomach is often associated with Helicobacter pylori (Hp) infection and Hp eradication therapy is widely accepted as initial treatment. The aim of this study is to report the clinical characteristics and long term outcome in a large series of patients with gastric MALT lymphoma exclusively treated with Hp eradication therapy.
Methods: 105 newly diagnosed gastric MALT lymphoma patients (54 men and 51 women) with median age of 64 years (range 20–94) referred to our institutions between June 1990 and November 2006 were eligible for the study. Staging was performed according to the Lugano staging system. All patients received anti-Hp eradication therapy as initial treatment with standard regimens combining antibiotics (usually Amoxicillin, Clarithromycin and/or Metronidazole) and proton-pump inhibitors (Omeprazole in most cases). Responses, evaluated with regular endoscopic biopsies every 3–6 months were graded according to the Wotherspoon’s histological score system.
Results: 100 patients had stage I and 5 patients had stage IIE1 disease. Hp was positive in 83 patients (79.5%) and was eradicated in all positive patients but 19 patients required a second line antibiotic therapy. Symptoms disappeared or markedly diminished and endoscopic features improved in almost all patients after Hp eradication. Histological regression of the gastric lymphoma was achieved in 78 of 102 evaluable patients (76%, 95% C.I.; 67%–84%) with histological complete response (Wotherspoon’s score 0–2) in 66 and partial response (score 3) in 12 patients. Of the 78 patients who achieved a lymphoma regression, the histological remission was consistently confirmed at follow-up endoscopies in 28 patients (36%), while 33 (42%) had histological score fluctuations (from 0–4), sometimes with transient histological relapses followed by spontaneous histological remissions. Ten patients had a frank lymphoma relapse (2 with high-grade transformation) and 7 died in remission for other causes. At a median follow up time of 6.3 years, the overall survival in the entire group is 92% at 5 years (95% C.I.; 84%–96%), 83% at 10 years (95% C.I.; 70%–91%) and 78% at 15 years (95% C.I.; 62%–88%). Only one patient died for lymphoma (after histological transformation). Additional tumors were observed in 22%. A history of autoimmune disease was present in 15% of the patients and did not affect the outcome.
Conclusions: Hp eradication usually results in long term disease control, independently of the lymphoma pathological remission status. A watch and wait policy seems safe in patients with local histological relapse with no endoscopic evidence of gross disease.