bluuebunny/crossref_metadata_2025_split huggingface dataset:
MDM2 Inhibitor Nutlin-3a Induces p53-Dependent Apoptosis Via Transcription-Dependent and -Independent Pathways and Overcomes Fludarabine-Resistance in CLL.
Authors: Kojima Kensuke , Konopleva Marina , Samudio Ismael J. , McQueen Teresa , Tsao Twee , Zeng Zhihong , Plunkett William K. , Andreeff Michael 
Published: October 2019

Abstract
Fludarabine containing combination therapies are the current standard for CLL therapy, in which p53-mediated induction of apoptosis contributes to leukemia cell killing. Although TP53 mutations occur in only 5–10% of patients with CLL, inactivation of the p53 pathway also occurs through Mdm2 overexpression or Atm deficiency. We investigate if recently developed potent and selective small-molecule antagonists of Mdm2, Nutlins, can overcome functional p53 inactivation associated with Mdm2 overexpression or Atm deficiency in CLL. Nutlin-3a caused a dose- and time-dependent increase in the percentage of Annexin V-positive cells in 20 primary CLL samples, irrespective of Mdm2 or Atm status. Samples with low Atm levels (n=3) were resistant to fludarabine. In addition to the transcriptional activation of target genes in the nucleus, cytoplasmic p53 can trigger transcription-independent apoptosis at the mitochondria. Samples with a cytoplasmic p53 localization pattern (n=7) showed a higher percentage of Nutlin-induced apoptosis than those with a nuclear pattern (n=8; P < .05). Furthermore, the inhibitory effect of cycloheximide pretreatment showed a negative correlation with the degree of Nutlin-induced apoptosis (r = - 0.617, P < .05). These findings suggest that the transcription-independent pathway may have stronger apoptogenic activity than the transcription-dependent pathway. The degree of Nutlin-3a-induced apoptosis directly correlated with apoptosis induced by the same concentrations (1–10 μM) of fludarabine in the early (24 hr) time period, and the Nutlin-3a/fludarabine combination induced synergistic apoptosis [averaged combination index (CI): 0.68]. Interestingly, this synergism was not affected by Mdm2 overexpression (CI: 0.59, n=12) or Atm deficiency (CI: 0.79, n=3). We conclude that (1) Mdm2 inhibitor Nutlin-3a efficiently induces p53-dependent apoptosis in wild-type p53 CLL cells, independent of Mdm2 overexpression or Atm deficiency, (2) transactivation-dependent apoptosis does not always play a major role in p53-dependent apoptosis and the exclusive activation of transactivation-independent pathway can fully induce apoptosis, (3) p53 activation plays a critical role in the early phase of fludarabine-induced apoptosis in vitro, and (4) Mdm2 inhibition and fludarabine synergistically induce apoptosis, which may overcome fludarabine-resistance in CLL.
Curcumin Enhances Differentiation of All-Trans Retinoic Acid (ATRA)-Sensitive and ATRA-Resistant Acute Promyelocytic (APL) Cells.
Authors: Kini Ameet R. , Nagabhushan Moolky , Tallman Martin S. , Roychowdhury Shantanu 
Published: October 2019

Abstract
The introduction of ATRA-based differentiation therapy has significantly enhanced outcomes in patients with APL. However, retinoic acid syndrome and ATRA-resistance remain significant concerns. It would therefore be useful to develop drugs that reduce the therapeutic doses of ATRA needed, and would be effective in ATRA-resistant cases. We have shown previously that curcumin, the yellow compound isolated from spice turmeric, suppresses the initiation and promotion stages of cancer development. In the present study we evaluated whether curcumin affects differentiation of NB4 APL cells. The NB4 cells were derived from a patient with APL, and differentiate in response to ATRA, while NB4-R1 cells are resistant to ATRA. Treatment of NB4 cells with 5 μM curcumin enhanced ATRA-mediated differentiation. Differentiation was assessed by evaluating CD11b expression, nitroblue tetrazolium (NBT) reduction and by morphologic examination. This curumin-mediated enhanced differentiation was apparent at 1 μM as well as 0.1 μM of ATRA. Curcumin alone did not cause differentiation of the NB4 cells, although higher concentrations of curcumin caused apoptosis. We then examined the effect of curcumin on the ATRA-resistant NB-R1 cells. Addition of ATRA and curcumin together induced differentiation of the NB4-R1 cells, whereas either agent alone did not cause differentiation. The differentiation was characterized by increased CD11b expression, NBT reduction and the typical morphologic changes. In addition, differentiation of the NB4-R1 cells was accompanied by restoration of the PML-oncogenic domains (PODs). These results indicate that curcumin may be another unconventional therapeutic agent in APL, following the successful use of ATRA and arsenic trioxide.
In Vitro Evaluation of Apoptosis and Cell Death of Neuroblastoma Cell Lines Exposed to Busulfan.
Authors: Kletzel Morris , Tallman Sarah C. , Olszewski Marie , Huang Wei 
Published: October 2019

Abstract
Objective: While busulfan is a commonly used chemotherapeutic agent in the treatment of many hematological diseases, its effectiveness against neuroblastoma is still in question. This study aims to assess the degree of apoptosis and cell death in neuroblastoma cell lines and primary neuroblastoma tumors when exposed to varying doses of busulfan.
Materials and Methods: Cultures from established cell lines SKN-SH, SKN-DOX-R, IMR-5, and NGP (n=4), as well as cultures from primary tumors (n=2) were seeded at 106 cells/ml in RPMI640 supplemented with 10% fetal bovine serum (FBS) and transferred to 24-well plates, where cells were exposed to 1ml of busulfan at 0, 0.001, 0.005, 0.01, 0.05, and 0.1mg/ml per well. Cells were incubated at 37°C in a humidified atmosphere of 5% CO2 for 72 hours. Wells were sacrificed after 0, 6, 24, 48 and 72 hours and tested with Annexin V and PI; 10,000 events were measured by flow cytometry. The percentage of apoptotic and dead cells was plotted in a graph and a t-test was performed against the untreated control.
Results: After 24 hours, there was a significant decrease in cell viability of each dose when compared to the control untreated cells (p<0.005).
24 Hour % Cell Viability for Varying Doses of Busulfan (mg/ml) Dose 0 Dose 0.001 Dose 0.005 Dose 0.01 Dose 0.05 Dose 0.1 Mean 66.1 44.4 40.3 40.7 37.7 39 SEM 5.56 5.17 5.96 6.17 6.03 5.60 Median 65 33.5 38 39 37 31 Range 39 to 97 14 to 87 4 to 89 6 to 93 4 to 77 5 to 88
The overall mean decrease in cell viability when compared to the control was 25.7%. However, there were only modest differences in effectiveness when comparing the doses, with an average of only 5–7% difference between doses. Further, there was much variability between the different cell lines, some with changes in apoptosis and cell death of over 50%, while other lines showed no changes at all. Limited differences were seen after 6 hours, and after 72 hours any effect of busulfan was masked by cell death due to other factors, as seen through increased cell death in untreated cells.
Conclusion: Busulfan induced apoptosis and cell death in vitro in neuroblastoma cell lines at a mean of 76.43% for non-resistant lines, 59.33% for primary tumors and 35% for resistant cell lines (at middle dose 0.01mg/ml). The resistance of certain cell lines confirms the difficulties of treating multi-drug resistant cells in often heterogeneous neuroblastoma tumors. That some cell lines were responsive shows the potential of using busulfan to treat neuroblastoma in the future.
A Novel Mechanism of Action of Methyl-2-cyano-3,12 dioxoolean-1,9 diene-28-oate (CDDO-Me): Direct Permeabilization of the Inner Mitochondrial Membrane To Inhibit Electron Transport and Induce Apoptosis.
Authors: Samudio Ismael , Konopleva Marina , Pelicano Helene , Huang Peng , Frolova Olga , Bornmann William , Ying Yun-ming , Evans Randall , Andreeff Michael 
Published: October 2019

Abstract
CDDO-Me is a synthetic oleanolic acid derivative that displays antitumorigenic and anti-inflammatory activities, and we have previously reported that this agent potently activates the intrinsic apoptotic pathway in leukemia cells. Here we demonstrate that mitochondrial dysfunction induced by CDDO-Me in leukemia cells is mediated by direct permeabilization of the inner mitochondrial membrane that results in the rapid depletion of mitochondrial glutathione (GSXm), loss of cardiolipin, and inhibition of mitochondrial respiration. More importantly, we demonstrate that in addition to activating the intrinsic apoptotic pathway, the mitochondrial effects of CDDO-Me may mediate its anti-inflammatory activity by modulating the generation of superoxide anion (O2−). Interestingly, CDDO-Me did not increase the generation of O2−, and in fact pretreatment of leukemia cells with CDDO-Me prevented the increase of this reactive oxygen species elicited by inhibition of complex I or III in the absence of de novo protein synthesis. CDDO-Me, but not other inhibitors of respiration, induced a time- and dose-dependent, cyclosporin A-independent permeability transition (mPT) of isolated mitochondria that was sensitive to sulfhydryl antioxidants but not to EDTA. PT induced by CDDO-Me and Ca2+ was accompanied by loss of GSXm suggesting that the increased permeability of the inner mitochondrial membrane facilitates the loss of this antioxidant. Finally, transmission electron microscopy revealed that CDDO-Me rapidly induced caspase-independent mitochondrial swelling and loss of inner membrane structure prior to the release of cytochrome c. Taken together, our results indicate that CDDO-Me is a novel antileukemic agent that induces apoptosis and inhibits mitochondrial electron transport via perturbations in inner mitochondrial membrane integrity.
Motexafin Gadolinium and Other Metallotexaphyrins Upregulate Heme Oxygenase-1.
Authors: Sirisawad Mint , Chen Jun , Ramos Jason , Miller Richard A. , Naumovski Louie 
Published: October 2019

Abstract
Heme oxygenase-1 (HO-1) is an inducible enzyme that is upregulated by heme and acts to degrade heme into bilirubin, carbon monoxide and iron. HO-1 is known to be upregulated in response to oxidative stress and has anti-apoptotic properties. Motexafin gadolinium (MGd, Xcytrin®) is a tumor-selective redox mediator that produces oxidative stress in cancer cells by reacting with various reducing metabolites and protein thiols to generate reactive oxygen species. Also, MGd is an expanded porphyrin that has a chemical structure which resembles heme. Since redox stress and heme upregulate HO-1 expression, we hypothesized that MGd would produce similar effects. We treated 8 hematopoietic tumor-derived cell lines with MGd and found that 2 of them (Ramos and HF-1) upregulated HO-1 protein. In Ramos lymphoma cells, we found that MGd induced HO-1 up to 8 fold by 24 hrs and that longer incubations did not result in appreciably greater amounts of protein. HO-1 levels returned to baseline in Ramos cells 48 hrs after cells were cultured in fresh media in the absence of MGd. HO-1 levels were elevated at baseline in HF-1 cells (compared to Ramos) and increased about 2 fold with MGd treatment. Other metallotexaphyrins, europium texaphyrin and dysprosium texaphyrin, also induced HO-1 expression. To determine if HO-1 expression was important for cell survival, we co-treated cells with MGd and tin protoporphyrin (SnPP), an inhibitor of HO-1 enzymatic activity. We found that both HF-1 and Ramos cells were sensitized to MGd-induced cell death by SnPP. These results demonstrate for the first time that metallotexaphyrins can upregulate HO-1 expression. The induction of HO-1 could be related to oxidative stress and/or metallotexaphyrins acting as heme mimetics.
Deletion of Any of BCR or ABL Gene on Derivative Chromosome 9 Is a Poor Prognostic Marker That Indicates Rapid Disease Progression in Chronic Myelogenous Leukemia.
Authors: Lee Young Kyung , Kim Young Ree , She Cha Ja , Yoon Sung-Soo , Kim Young Soo , Cho Han Ik , Kim Hee Chan , Lee Yun Song , Lee Dong Soon 
Published: October 2019

Abstract
To evaluate the prognostic significance of the submicroscopic deletions of ABL or BCR gene associated with t(9;22) in CML, we investigated the incidence of ABL or BCR deletion on derivative chromosome 9 using fluorescent in situ hybridization (FISH), and analyzed the survival. FISH was performed using LSI BCR/ABL, dual fusion translocation probe on bone marrow cells of 86 patients with CML. Of 86 patients, ABL deletion was detected in 13 (15.1%) patients and BCR deletion in 8 patients (9.3%). Patients with ABL deletion showed shorter event-free survival time (EFS) than those without ABL deletion (P=0.020). Patients with BCR deletion showed significantly short overall survival time (OS) (P=0.039). Patients with ABL and/or BCR deletion (14/86 patients, 16.3%) showed significantly short OS and EFS (median OS: 43.0 months, median EFS: 40.0 months), compared to the patients without any deletions of BCR or ABL gene (median OS: 94.0 months, median EFS: 90.0 months)(P=0.041 for OS, P=0.008 for EFS). Patients with BCR deletion, all except one had a concomitant ABL deletion, suggesting that BCR deletion occurs in conjunction with ABL deletion. In patients with ABL deletion only, BCR/ABL rearrangement with b2a2 mRNA type tended to be more frequent than in patients without any deletion of the two genes (P=0.073). Deletion of any of BCR or ABL gene on derivative chromosome 9 was associated with both short OS and EFS. We conclude that deletion of not only ABL gene, but also BCR gene is a poor prognostic marker that indicates rapid disease progression in CML.
Study on Clinical and Biological Characteristics of Childhood Acute Leukemia with MLL Gene Rearrangements.
Authors: He Jun , Chen Zi-xing , Xue Yong-quan , Pan Jin-lan , He Hai-long , Li Jian-qin , Wu Ya-fang , Huang Yi-ping , Zhu Ling-li 
Published: October 2019

Abstract
The rearrangement of MLL gene is reported in 70%~80% of infant and in 5%~10% of older children (under the age of 15) with acute leukemia (AL). The biological features associated with alterations in MLL gene are hyperleukocytosis, CD10−/CD19+ phenotype and very poor prognosis. To explore the MLL rearrangement in details in our AL children patients and obtain more information on the relationship between the MLL gene abnormality and clinical outcomes. The following study has been conducted. A total of 298 patients with AL attended The Affiliated Children’s Hospital of Soochow University, including 16 cases with MLL rearrangements, were recruited in this study. Of the cohort, 11 were diagnosed as ALL, 5 were AML. 9 of 16 patients were in infant age (up to 2 year) and the rest were between the age of 2 to 13 years. Fluorescence in situ hybridization (FISH) analysis using LSI MLL dual color probe. Multiplex reverse transcriptase- polymerase chain reaction (multiplex RT-PCR) were used to discriminate 13 different fusion transcripts. These results were analyzed together with R banding karyotyping and immunolphenotyping determined by flow cytometry. We have found MLL rearrangements in 16 cases of childhood AL which were accounted for 5.4% of 298 AL patients, and 56.3% of infant AL. Among 106 cases analyzed by multiplex RT-PCR, MLL gene rearrangement was found in 11 cases, including MLL/AF4 fusion gene in 2, MLL/AF6 fusion gene in 1, MLL/AF6, MLL/ELL combined with MLL/AFX or HOX11 in one of each, MLL/AF9 in 2, MLL/AF10 in 1, MLL/ELL in 2. MLL partial tandem duplication in 1. In addition an activated HOX11 gene was found in 1 case.. In 27 cases assayed by FISH, MLL gene rearrangements have been detected in 9 cases (36.0%). In 16 patients with MLL gene rearrangements, 14 (87.5%) exhibited clonal chromosome abnormalities involved chromosome 11 in 11 cases, presenting as t(4;11) in 2, t(6;11), t(8;11), t(7;8;11), and t(9;11) in one of each, respectively, trisomy 11 in 2 and 11q- in 3 cases. Among these 16 patients, 11 were B-ALL, including Pro-B and Pre-B ALL; 5 of AML-M5, 3 of these 5 M5 patients were CD7+ and CD2+. Of these 16 patients 8 received chemotherapy and 7 of them achieved complete remission, while the other 8 patients eventually gave up treatment. Our results demonstrated that multiplex RT-PCR combined with FISH provided a more accurate and sensitive method for detection of MLL gene rearrangements, including chromosome translocation, deletion and duplication. Our findings lead to the detection of novel rearrangements at molecular genetic level. These findings regarding the MLL rearrangement provide most important information in guiding therapy and predicting prognosis in childhood AL. Besides our results also provide evidence in support of the value of 11q23/MLL in WHO classification categories.
Prospective Multicentric Molecular Study for Poor Prognosis Fusion Transcripts at Diagnosis in Adult ALL Patients - The LALA94 Experience.
Authors: Gabert Jean A. , Picard Christophe , Hayette Sandrine , Bilhou-Nabera Christelle , Cayuela Jean-Michel , Macintyre Elizabeth , Frenoy Nicole , Preudhomme Claude , DuPont Madeleine , Bastard Christian , Bories Dominique , Vaerman Jean-Paul , Davi Frédéric , Dastugue Nicole , Raynaud Sophie , Lafage Marina , Fest Thierry , Gaub Marie-Pierre , Lheritier Véronique , Thomas Xavier , Charrin Christiane , Boucheix Claude , Dombret Hervé , Fiere Denis 
Published: October 2019

Abstract
From 1994 to 2000, 984 adults aged from 15 to 55 years with newly diagnosed Acute Lymphoblastic Leukemia (ALL) were eligible for randomization in the multicentric LALA-94 clinical protocol. The t(9;22), t(1;19) and t(4;11) translocations corresponding to BCR-ABL, E2A-PBX1 and MLL-AF4 fusion gene transcripts respectively, were considered as independent poor prognostic factors. Standardized RT-PCR analysis of these fusion gene transcripts were performed by 17 laboratories in order to provide data before the second randomization (J35) on 787 patients. In this multicentric study, validated data were available for therapeutic stratification for 91% of these analysed patients. No false positive RT-PCR was reported. Secondarily to retrospective BCR-ABL FISH, few false BCR-ABL negative RT-PCR were identified, leading to the design of new BIOMED-1 primers for b3-a3 junctions detection. Moreover, the LALA-94 study allowed to define new guidelines for molecular analysis at diagnosis.
Like in other studies, the BCR-ABL transcript was found to be the most frequent molecular abnormality in B-ALL (24%) whereas MLL-AF4 and E2A-PBX1 were detected in 5% and 3.5% of B-ALL, respectively. Epidemiological and clinical data of MLL-AF4 and E2A-PBX1 were concordant with previous publications. Interestingly, because of the large number of reviewed patients, the different BCR-ABL subtypes (M-BCR and m-BCR) were statistically characterized by few clinical data.
M-BCR subgroups had a higher age than m-BCR (p= 0.016) and occurs especially during the second semester (p= 0.034). Moreover, the comparison of clinical data at diagnosis of M-BCR variants showed that median age of b3a2 was statistically younger than b2a2 (p= 0.04) and that b3a2 occurs more frequently in man (p= 0.02). For the first time, these data suggest that these BCR-ABL breakpoints: m-BCR and M-BCR and also b2a2 and b2a3, are secondary to different physio-oncologic mechanisms even if therapeutic regimens including the same targeted therapy (tyrosine kinase inhibitor) for all BCR-ABL variants is the rule today.
FOXP3 Expression in B and T Cell Lymphomas.
Authors: Roncador Giovanna , Garcia Juan Fernando , Garcia Jose Francisco , Maestre Lorena , Lucas Elena , Cigudosa Juan Cruz , Sanchez Lydia , Ohshima Koichi , Nakamura Shigeo , Banham Alison , Piris Miguel Angel 
Published: October 2019

Abstract
Foxp3, which encodes a forkhead/winged helix transcription factor designated Scurfin, is a key regulatory gene required for the development and function of regulatory CD4+CD25+ T cells (Treg), a subpopulation of T-cells specialized in maintaining the balance between immunity and tolerance.
Humans with defects in the FOXP3 gene, develop strong activation of the immune system, leading to multiorgan autoimmune disease, allergies, inflammatory bowel disease and severe infections, collectively known as the IPEX syndrome (immune deregulation, polyendocrinopathy, enteropathy, X-linked inheritance syndrome)
Because of the importance of FOXP3 in the development and function of Treg cells, and its potential use as a specific Treg marker, we have developed several monoclonal antibodies against FOXP3, for use on paraffin-embedded tissue sections and evaluated its expression in a large series (150 cases) of B- and T-cell lymphomas. In reactive lymphoid tissue, strong nuclear FOXP3 expression was observed in approximately 5% of interfollicular T-cells. FOXP3 expression in tumour cells was confined to most of Adult T-cell Leukaemia/Lymphoma (ATLL) cases (68%), with some variability in protein expression. In other lymphoma types, FOXP3 expression was only detected in the reactive T-cell background, and the number of FOXP3-positive reactive T-cells was variable, ranging from almost a complete absence (Burkitt’s lymphoma) to abundant infiltrate (common in follicular lymphoma).
In conclusion, the availability of a FOXP3 monoclonal antibody, not only provides an important tool for the study of the development and function of Treg cells, but also represents a useful marker for the identification of ATLL cases in formalin-fixed paraffin-embedded tissue sections.
The presence or absence of Treg cells in the tumour environment could also play a role in the immune surveillance of tumours, thus implying a potential additional value for the detection of this cell population in tumour samples.
Quantitative Comparison of BCR/ABL Transcript Levels in Bone Marrow and Peripheral Blood Specimens from Patients with Minimal Residual Disease.
Authors: Faulkner Nicole E. , Sirko-Osadsa D. Alexa , Rosenblum-Vos Lynne S. , Rohlfs Elizabeth M. , Browne Patrick , Hallam Stephanie E. 
Published: October 2019

Abstract
Chronic Myeloid Leukemia (CML) is diagnosed by the presence of the Philadelphia chromosome (Ph+). The number of cells containing the bcr/abl chromosomal translocation diminishes dramatically after current standard of care treatment. Patient progress is monitored quantitatively and long-term prognosis is favorable if the Ph+ cells are no longer detected by cytogenetic analysis or FISH (sensitivity of 20% and 2% respectively). Molecular monitoring of cDNA BCR/ABL transcripts is widely used to assess levels of minimal residual disease (MRD). In our laboratory, quantitative RT-PCR allows detection of BCR/ABL transcript down to 1 x 10−5 or one BCR/ABL transcript in 100,000 reference gene transcripts. Serial values from bone marrow (BM) and blood specimens are tracked over time and monitored for increases that may indicate unfavorable changes in disease status. Though the literature indicates that the levels of BCR/ABL transcript in BM and blood samples are comparable at the time of diagnosis, this may not be true in patients with MRD. To explore this question we compared BCR/ABL transcript levels in paired BM and blood samples that were either collected together or separately but within a short time frame. In 36% of these cases both specimen types had detectable low-level BCR/ABL transcript. The levels of transcript in bone marrow were consistently higher than the levels in the associated blood specimen, though there was a wide range of variability (3-50 fold). Additionally, when blood levels of transcript were near the lower limit of detection, we were more likely to observe a larger difference relative to bone marrow transcript levels than when blood levels of transcript were higher. These results indicate that while both BM and blood specimens are valuable for serial molecular monitoring, they may not be equivocal when sampled from patients with MRD. If a patient has been monitored using blood specimens, periodic concurrent testing of blood and bone marrow specimens can provide clinical utility.