bluuebunny/crossref_metadata_2025 huggingface dataset:
FGFR3 Tyrosine Kinase Inhibitor AB1010 as Treatment of t(4;14) Multiple Myeloma.
Authors: Arnulf Bertrand , Ghez David , Leblond Veronique , Choquet Sylvain , Belhadj Karim , Macro Margaret , Bouscary Didier , Facon Thierry , Moreau Philippe , Jaccard Arnaud , Soulier Jean , Karlin Lionel , Hermine Olivier , Fermand Jean-Paul
Published: October 2019

The t(4;14)(p16;q32), found in 15% of multiple myeloma (MM) cases, is associated with a short time to progression (TTP) despite a good initial response. In this subset of MM, relapses are resistant to conventional and intensive chemotherapy leading to a poor prognosis. Plasma cells with t(4;14) ectopically express the fibroblast growth factor receptor 3 (FGFR3), a tyrosine kinase receptor which has proven transforming activity and may represent a therapeutic target. We have studied the safety and efficacy of AB1010, an FGFR3 tyrosine kinase inhibitor, in patients with relapsing/refractory t(4;14) MM. 24 MM patients (M 33%, F 67%, median age 55 years) with t(4;14) were enrolled. FGFR3 expression was detected in all but one patient. AB1010 (9 mk/kg/d) was given orally and Dexamethasone (Dex) (40 mg/d X4d/month) was added if progression. In case of explosive relapse (defined by deep cytopenia, renal failure, circulating plasma cells), a chemotherapy was given followed by a wash out period of 1 month before start of AB1010. Two patients were enrolled after the first line of treatment at a plateau state with a response < 75% (<75% decrease of the monoclonal component). Among the 24 patients, 19 were evaluable since 5 had a duration of treatment < 1month. The main toxicities were gastrointestinal (nausea 63,5%, grade I/II, diarrhoea 25%, grade I; anorexia 25%, grade II) and oedema (face 50%, grade I/II and legs 24% grade I). Hematological toxicities were limited to a transient grade III neutropenia in one patient. Dex was added in all cases. Six patients (3 in 1st relapse, 3 in >3rd lines) had explosive relapse. Treatment regimen before start of AB1010 were :VTD (4); Thal/Dex (1); MLPHD (1). In these 6 cases, the TTP range from 1,5 to 4 months. 11 patients had non explosive relapse. Of these, 5 were in 1st relapse in whom 1 near CR (figure) and 1 PR were observed, with a median TTP of 10 months. Six patiens were in > 2nd relapse and 2 MR were obtained. Of the 2 remaining patients enrolled at plateau, 1 progressed at 6 and 1 is still on going after 13 months. In conclusion, AB1010, in combination with Dex, is well tolerated and may be useful in the treatment of patients with t(4;14) MM, especially in early phase of the disease. Responses were observed (40%) in patients with first relapse with a TTP superior as compared to the reported one (4,7 months with Thalidomide/Dex, Jaksic et al, 2006). Given the synergistic effect on MM cells proliferation observed in vitro, a study is now on going to test the safety and the efficacy of the combination AB1010 + Dex + Velcade in early relapsing patients with t(4;14) MM.

Figure Figure
SMRT; Not So Smart in Multiple Myeloma.
Authors: Ghoshal Pushpankur , Houde Christiane , Szafranek Angela , Nganga Alain , Johnson Timothy , Bigelow Ashley , Moran-Guiati Joseph , Smiraglia Dominic , Chanan-Khan Asher Alban , Coignet Lionel J.
Published: October 2019

Multiple myeloma (MM), a clonal B-cell malignancy, characterized by accumulation of plasma cells in bone marrow is the second most hematological malignancy in United States. In the BM, the myeloma and stromal cells secrete cytokines, which support the growth and survival of the myeloma cells. We previously showed that JAG2, one of the NOTCH ligand, is over-expressed in MM cells (Blood. 2004 Dec 1;104(12)). We hypothesized that over-expression of JAG2 in myeloma cells induce secretion of IL-6 from the stromal cells and subsequently enhances the proliferation of myeloma cells. JAG2 has been shown to be over-expressed in all cell lines and patient samples studied To identify the mechanism for JAG2 overexpression in MM cells, we assessed the potential modifications of the JAG2 promoter in MM cell lines as well as patient samples (an JAG2 negative controls) by studying both methylation and histone acetylation level of the JAG2 promoter. We show that difference in H4 acetylation level might play a crucial role in JAG2 expression. Acetylation state of histones can be regulated by the recruitment of histone deacetylases (HDACs). HDACs are typically recruited to promoter regions through interaction with nuclear co-repressors such as SMRT. The cell lines and patient samples studied presented significantly reduced levels of SMRT. Therefore, based on these observations we propose a model in which partial down-regulation of SMRT recruits less active HDAC3 (as confirmed by immunoprecipitation). As a result the deactylation process of histones in the JAG2 promoter region has been impaired and the cells lost their regulatory mechanism on transcriptional regulation of JAG2. This provides a mechanistic explanation for JAG2 over-expression in MM and on the direct involvement of the SMRT co-repressor in MM pathogenesis.
Insertional Activation of GLI2 in Adult T-Cell Leukemia/Lymphoma.
Authors: Cleveland Susan M. , Dave Utpal P.
Published: October 2019

Retroviruses induce cancer by integrating into the cellular genome and activating oncogenes or inactivating tumor suppressor genes. Human T-cell Leukemia Virus type 1 (HTLV-1), a complex retrovirus, induces Adult T-cell Leukemia/Lymphoma (ATLL) after a latency of over 30 years and in only 5% of carriers. The long latency and incomplete penetrance is similar to how slow transforming retroviruses induce cancer in mice and imply multiple oncogenic “hits” need to accumulate for clinically apparent disease. Insertional mutagenesis may be one mechanism by which ATLL develops. We used splinkerette-PCR to clone and map insertion sites from an HTLV-1 infected T-cell line, Hut-102. We identified an HTLV-1 insertion 5′ of the GLI2 gene, formerly known as Tax-Helper-Protein-1. We found GLI2 was up-regulated by promoter insertion. Interestingly, we found GLI2 protein occupied the HTLV-1 Long Terminal Repeat. The effect of GLI2 expression on viral expression was investigated by knockdown of GLI2 in Hut-102 cells. Our results show that retroviral insertional mutagenesis can be an important mechanism in HTLV-1-induced leukemias and lymphomas.
Mechanism of Tetrandrine in Combination with Droloxifen To Reverse Multidrug Resistance in Leukemia.
Authors: Chen Bao-An , Su Ai-ling , Gao Feng , Cheng Jian , Ding Jia-Hua , Gao Chong , Sun Yun-yu , Wang Jun , Zhao Gang , Chen Ning-Na , Zhao Hui-Hui , Pei Xiao-Ping
Published: October 2019

Objective: The study aims to investigate the effect of Tetrandrine (Tetrandrine, Tet) in combination with Droloxifen (Droloxifen, DRL) on the expression of NF-κB(nuclear factor kappaB) of K562 and K562/A02 cell lines and the reversal mechanism of this combination.

Method:

The activation of NF-κB in K562 and K562/A02 cell lines and the effect of Tet,DRL alone or in combination on NF-κB were determined with Immunocytochemistry and Western blotting respectively.

Results:

K562/A02 cells displayed higher level of NF-κB protein expression than their parental K562 cells; The application of Tet or DRL alone or in combination had no effect on NF-κB expression in K562 cells at 6h and 12h; Tet and DRL used alone or in combination could significantly down-regulate NF-κB protein expression in nucleaus of K562/A02 cells.

The effect was more significant in combination of Tet and DRL than the application of single drug. This decrease became more significant at 12h than at 6h.

Conclusions:

Activation of NF-κB may be involved in the mechanism of MDR of K562/A02 cell line; Inhibition of NF-κB activation may be involved in the mechanism of multidrug resistance reversal to K562/A02 line of Tet and DRL.
L-Gossypol Has Significant In Vitro Activity Against Primary Acute Myeloid Leukaemia Cells, Inducing Apoptosis through the Intrinsic Pathway.
Authors: Osman Hani Y. , Knapper Steven , Burnett Alan K.
Published: October 2019

The potential anti-tumor effect of the polyphenolic cotton oilseed extract gossypol has been known for a number of years. Studies in cell lines and primary tumor material have suggested both antiproliferative effects and the induction of apoptosis (Huang et al. Anticancer research 26:3a) with activity being seen against even multidrug resistant tumors. Although the mechanism of action of gossypol has not yet been well-characterised, there is some evidence that it may block the binding of the apoptotic regulator BCL-2 with the pro-apoptotic proteins BAX and BAD (Prada et al. blood 106:11). Acute myeloid leukaemia (AML) is characterised by accumulation of primitive blasts in bone marrow and peripheral blood. The disease is highly heterogeneous and inherently resistant to therapy, dysregulation of apoptosis frequently leading to a survival advantage in blast cells. We studied the in vitro activity of gossypol against cryopreserved primary blasts obtained from 44 AML patients. Cells were incubated for 48 hours with a range of concentrations of gossypol, this being followed by MTS cytotoxicity assay. IC50 concentrations ranged from 792nM to in excess of 100 mM (median 22.791 mM). Statistical analysis showed no significant variation in drug sensitivity according to sex, age, presenting white cell count, FLT3 mutation status, karyotype or clinical outcome measures. The in vitro combination of gossypol with the cytotoxic agent cytosine arabinoside (AraC) was assessed in 40 primary samples using the median effects principle of Chou and Talalay. The median combination index of 0.946 suggested an overall additive effect when the agents were combined, although numbers were insufficient to establish this conclusively (95% CI for median 0.84–1.31). Flow cytometric annexin V binding assay confirmed apoptosis to be the principal mode of cell death in response to gossypol, cell viability obtained using this method correlating well with that obtained using the MTS assay. Gossypol caused downstream activation of Caspase 3/7 via Caspase 9 as determined by luminometric analysis, indicating that the intrinsic pathway is the main inducer of this cycle. Gossypol is effective in inducing apoptosis in primary AML blasts and there is potential for greater efficacy when used in combination with cytotoxic chemotherapy. Further in vitro development of this agent is now required to further define its future clinical role.
Metalloproteinase Inhibitors Increase the Survival of Long-Term Refrigerated Platelets in Mice.
Authors: Josefsson Emma C. , Rumjantseva Viktoria , Dahlgren Claes , Bergmeier Wolfgang , Wagner Denisa D. , Hartwig John H. , Hoffmeister Karin M.
Published: October 2019

The αMβ2 receptor on macrophages, which recognizes clustered β-N-acetylglucosamine (βGlcNAc) residues on glycoprotein (GP) Ib-V-IX, is one phagocytic receptor involved in the removal of transfused refrigerated platelets. Although coverage of the exposed βGlcNAc residues by galactosylation prevents the clearance of murine platelets refrigerated for less than 4h, it does not impact the clearance of platelets refrigerated for 48h, suggesting that further changes occur in platelets during prolonged refrigeration. We have now found that murine GPIbα and GPV are partially shed from the platelet surface after refrigeration for 48h and re-warming. Other members of the complex, GPIX and GPIbβ, were not affected. Platelets isolated from mice with inactive ADAM17 enzyme, a GPIbα and GPV sheddase, had preserved surface GPIbα and decreased GPV shedding compared to WT mice after long-term refrigeration. 48h refrigerated platelets with inactive ADAM17 or platelets treated with the metalloproteinase inhibitor GM 6001, had increased survival when transfused into WT mice. Metalloproteinase inhibitor-treated and galactosylated, 48h refrigerated and re-warmed mouse platelets exhibited survivals equivalent to fresh platelets. We conclude that 48h long-term refrigeration and re-warming of platelets induces shedding of GPIbα and GPV in murine platelets, and that metalloproteinase inhibitors increase the survival of long-term refrigerated platelets in mice. Further changes in GPIb-V-IX in long-term refrigerated platelets may recruit additional phagocytic receptors to promote clearance, in addition to the αMβ2 receptor.
The Expression and Clinical Significance of PRAME Gene in Acute Leukemia.
Authors: Fu Rong , Ding Kai , Shao Zonghong
Published: October 2019

Objective To investigate the expression of PRAME (preferentially expressed antigen of melanoma) gene in acute leukemia and its clinical significance in monitoring prognosis, detecting minimal residual disease (MRD) and gene immunotherapy.

Methods The expression of PRAME gene mRNA in bone marrow mononuclear cells is measured by reverse transcriptase polymerase chain reaction in 34 patients with acute leukemia and 12 bone marrow samples of health donors. The relationships between PRAME gene expressions and some clinical data, such as gender, age, white blood count, leukemic immunophenotype, the percentage of blast cells, and the karyotype of chromosome, were also estimated.

Results PRAME gene was expressed in 38.2% of all the patients, 40.7% of all the AML patients, which was higher than the 28.6% of ALL patients (p >0.05). There was no expression of PRAME gene in healthy donors. In all the sub phenotypes of AML, the expressive rate of PRAME gene in M3 patients is 80%, which is higher than that in M2 (33.3%) and in M5 (28.6%). The expressive rate of PRAME gene was also positively correlated with the expression of CD15, CD33, and the abnormality in the karyotype of chromosome, but not correlated with age, gender, white blood count and percentage of blast cell in bone marrow.

Conclusion PRAME gene is highly expressed in acute leukemia, and could be regarded as a useful tool for monitoring MRD. Differential expression in acute leukemia patients vs. healthy donors suggests that the immunogenic antigens PRAME are potential candidates for immunotherapy in acute leukemia.
Prognostic Impact of Cytogenetic Abnormalities in Elderly Patients with Acute Myeloid Leukemia (AML) Enrolled in the ALFA-9803 Trial.
Authors: Terré Christine , Gardin Claude , Gachard Nathalie , Tigaud Isabelle , Désangles François , Bastard Christian , Maarek Odile , Lai Jean-Luc , Bernheim Alain , Perot Christine , Nguyen-Khac Florence , Eclache Virginie , Plessis Ghislaine , Turlure Pascal , Thomas Xavier , de Revel Thierry , Fenaux Pierre , Dombret Hervé
Published: October 2019

We have recently reported the superiority of prolonged ambulatory versus short intensive post-remission therapy in 416 patients with AML aged 65 years or more (median age, 72 years) selected to be treated intensively in the ALFA-9803 trial (C. Gardin et al. Blood 2007). We report here on the prognostic value of cytogenetic abnormalities in 339 of these patients with available karyotype (27 not done, 50 failures). When classical endpoints (CR rate, DFS, OS) were used, we were only able to identify one very unfavorable subgroup of 14% patients (N=49 with −7, abnormalities of both chromosomes 5 and 7, 3q abnormality, or complex karyotype with 5 abnormalities or more; CR rate, 30%). No significant differences in outcome were found between patients with any other abnormality and the 171 patients with a normal karyotype/−Y (overall CR rate, 62%). When cumulative incidences of AML-related events (primary refractory AML, relapses) were used as endpoints with induction deaths and deaths in first CR as competing events, three distinct subsets were more easily identified:

14% very unfavorable-risk patients, as detailed above; 59% standard-risk patients (N=200 with t(8;21), inv(16), normal karyotype/−Y, or isolated +8; CR rate, 65%); and 27% unfavorable-risk patients (N=90 with other abnormalities, including notably 5q− and 7q−; CR rate, 58%).

Median failure-free interval (Fig 1; P<.001, by the Gray test) and relapse-free interval (Fig 2; P<.001, by the Gray test) were 2.3, 12.4, and 8.1 months and 4.2, 15.0, and 10.5 months in these three subsets, respectively. Interestingly, prolonged ambulatory post-remission therapy was associated with longer DFS and OS in the standard-risk subset only. The results demonstrate that despite their poor general outcome, older patients with AML may be subdivided in three meaningful cytogenetic subgroups. This 3-subgroup classification should help to optimize disease management decisions such as intensive versus non-intensive or investigational therapies for induction or consolidation.

1. Failure-free interval 1. Failure-free interval 2. Relapse-free interval 2. Relapse-free interval
Flow Cytometric Examination of Non-Hematic Body Fluids in Hematologic Malignancies: A Comparison with Cytology.
Authors: Cesana Clara , Scarpati Barbara , Brando Bruno , Barba Claudia , Ferri Ursula , Scampini Linda , Oreste Pierluigi , Veronese Silvio , Nosari Annamaria , Morra Enrica , Cairoli Roberto
Published: October 2019

Introduction: Flow cytometry is well known to detect malignant cells in peripheral blood and bone marrow of patients with hematologic malignancies. However, its role in evaluating non-hematic body fluid contamination by tumor cells is largely unexplored.

Patients and Methods: Data detected by flow cytometry in non-hematic body fluid samples drawn between 2002 and 2007 from patients with hematologic neoplasms were retrospectively compared with morphological findings obtained from cytospin slides. Immunophenotyping was carried out by using disease-specific multicolor panels of quadruple monoclonal antibodies, conjugated with the fluorochromes FITC, PE, PerCP, and APC, respectively. Acquisition of information on 1x104 to 1x105 stained cells depending on the whole sample cellularity was assessed on a dual-laser FACSCalibur flow cytometer using the CellQUEST software (Becton Dickinson, San José, CA, USA).

Results: Fourty-five samples (bronchoalveolar fluid, n=5; ascites, n=2; hydrocele, n=2; pleural effusion, n=8; and cerebrospinal fluid, n=28) from 32 patients were available for comparison. Diagnoses were as follows: chronic myelomonocitic leukemia (n=1), acute promyelocytic leukaemia (n=2), acute myelomonocytic leukaemia (n=1), B-chronic lymphocitic leukemia (n=4), follicular lymphoma (n=1), acute lymphoblastic leukaemia (n=6), lymphoblastic lymphoma (n=1), high grade non-Hodgkin’s lymphoma (NHL), Burkitt-like (n=3), diffuse large B-cell NHL (n=6), peripheral T-cell NHL (n=3), and NHL, unspecified (n=4). Flow cytometry detected neoplastic cells in 24 cases. Of these cases, only 17 were positive also by morphology. In 7 cases, in which tumor cells were detected by flow cytometry but not by morphology, clinical data confirmed the presence of the disease. Flow cytometry did not show neoplastic cells in 21 cases. Of these cases, only 18 were negative also by morphology. In the remaining 3, the suggestion of diffuse large B-cell NHL contamination by morphology was not confirmed by flow cytometry, demonstrating T-reactive lymphocytes that were clearly negative for disease-specific markers.

Conclusions: Our data suggest that flow cytometry is a useful tool complementary to morphology for the screening of non-hematic body fluid contamination in patients with hematologic neoplasms.
High-Resolution Analyses of Genetic and Epigenetic Aberrations in Infant Leukemia with MLL Rearrangement.
Authors: Takita Junko , Kato Motohiro , Nakamura Fumihiko , Chen Yuyan , Yamamoto Go , Nannya Yasuhito , Sanada Masashi , Taki Tmohiko , Kikuchi Akira , Igarashi Takashi , Hayashi Yasuhide , Ogawa Seishi
Published: October 2019

MLL rearrangement-positive leukemia is one of the most aggressive types of leukemia. It is diagnosed predominantly in infants and typically shows a multilineage phenotype. Since current chemotherapy fails in more than 50% of infantile leukemia with MLL rearrangement, a better understanding of biological features of the disease is importantly in order to develop more specific and successful treatment strategies. Thus, to explore both genetic and epigenetic lesions associated with MLL rearrangement-positive infantile leukemia, we performed genome-wide analyses of copy number alterations/allelic imbalances as well as methylation analysis using Affymetrix GeneChip SNP genotyping microarrays and promoter tiling array combined with methylated DNA immunoprecipitation (MeDIP). Combined with newly developed algorithm, CNAG/AsCNAR, SNP array analysis enables accurate copy number analysis at extremely high resolutions. In addition, by sensing subtle distortions in allele-specific signals caused by allelic imbalance using anonymous controls, sensitive detection of LOH is enabled with accurate determination of allele-specific copy numbers even in the face of up to 70–80% normal cell contamination. In total, 25 specimens from MLL rearrangement-positive leukemia were analyzed using high-density SNP-genotyping microarrays (Affymetrix GeneChip 100K/500K arrays). While unbalanced translocation involving chromosome 11q23 is the most frequent genetic alterations, uniparental disomy of 17q is also common genetic alterations in MLL rearrangement-positive leukemia. A number of other genetic changes were also identified but these were mostly found in a single case. On the other hand, epigenetic abnormalities have been implicated in MLL rearrangement-positive leukemia, because MLL is known to be involved in epigenetic regulations. In our assay, fragmented genomic DNA from leukemia specimens was immunoprecipitated with anti-methylcytosine antibody (MeDIP). The immunoprecipitated DNA was amplified by PCR and subjected to hybridization to the promoter tiling array, in which regulatory regions of more than 25,000 genes are tiled by 6.5 millions of oligonucleotide probes to enable sensitive detection of target sequences. This tiling array covers approximate 59% of CpG islands in the human genomes. In addition to the previously described genes, such as FHIT and HOX family, a number of tumor specific methylated sites were identified in the leukemic cells and which were subsequently verified by bisulfate sequencing. These results indicated that high-resolution analyses of genetic and epigenetic aberrations using microarray techniques are powerful and useful for detection of new findings in the pathogenesis of infant leukemia with MLL rearrangement.
Whole Genome Association Study in Acute Myeloid Leukemia with a Normal Karyotype, Using a Single-Nucleotide Polymorphism (SNP) Analysis.
Authors: Cha Yongjun , Ahn Kwang-Sung , Park Juwon , Kim Byung-Su , Bang Soo-Mee , Kim In-Ho , Yoon Sung-Soo , Lee Dongsoon , Lee Jong Seok , Lee Young Yiul , Park Seonyang , Kim Byoung Kook
Published: October 2019

As a result of numerous genetic aberrations, acute myeloid leukemia (AML) present with very heterogeneous clinical features. Moreover, structural and numerical chromosome aberrations currently comprise the most important basis for predicting these heterogeneous outcomes. However, 30 to 50 percent of AML patients have cytogenetically normal karyotypes. Though submicroscopic genetic alterations (such as NPM1, CEBPA, MLL-PTD) are increasingly being used for clinical purposes, additional markers with prognostic or predictive value are still lacking in this group. In this study, we performed a genome-wide, single-nucleotide polymorphism (SNP) study in order to identify novel genomic regions of interest in normal karyotype AML. 54 untreated AML patients with normal karyotypes were analyzed with an Illumina infinium 317K SNP chip assay. SNP genotype call rate was 99.8 percent, resulting in a resolution of 1 SNP/Mb. In a genome-wide SNP analysis, 317 SNP loci, having a significant difference between AML and normal control, were found. In addition, about 300 loci were also identified, showing a significant difference between patients who achieved CR and those who did not. Some of these were found to be related with drug metabolism and MDR family. Also we used the SNP assay to screen a loss of heterozygosity (LOH), suggesting possible involvement of tumor suppressor genes. In summary, 38 LOH regions larger than 1Mb were observed in 23 cases (43%) among 54 AML patients having normal karyotypes. Most (55%) of them were copy-neutral events and the most frequently identified alterations were located at 3p, 8p, 13q and 22q. 11 (29%) out of 38 LOH regions had overlapping parts among them. Some LOHs were correlated with response to chemotherapy. Various genetic changes were discovered by use of an SNP-based chip array in normal karyotype AML patients. Moreover, some of these were related with CR acquisition and drug metabolism. These results can be used to differentiate normal karyotype AML and warrant further study.
Essential Role of Pro-Apoptotic Mechanisms for Production of Normal Erythrocytes and Prevention of Hemolysis.
Authors: Chen Min , Prchal Josef T. , Hector Sandoval , Thiagarajan Perumal , Dasgupta Swapan K. , Schumacher Armin , Wang Jin
Published: October 2019

Prevention of apoptosis is a well-recognized mechanism essential for productive early erythropoiesis and preventing anemia; however, the importance of pro-apoptotic processes for productive terminal erythropoiesis is not defined. Erythroid cells undergo enucleation and the removal of organelles during terminal differentiation. Although autophagy has been shown to mediate the elimination of organelles for erythroid maturation, the molecular mechanisms underlying this process remain unknown. Here we report a novel role for Bnip3L, a pro-apoptotic Bcl-2 family member, in the regulation of erythroid maturation through autophagy. Bnip3L−/− mice developed hemolytic anemia and expansion of erythroid precursors (Fig.1A). Moreover, erythrocytes in the peripheral blood of Bnip3L−/− mice exhibited mitochondrial retention associated with reduced lifespan (Fig. 1B). While the clearance of ribosomes proceeded normally in the absence of Bnip3L, the entry of mitochondria into autophagosomes was defective (Fig. 1C). Overexpression of Bnip3L led to the loss of mitochondrial membrane potential (ΔΨm). Further, disrupting Δ Ψ m with an uncoupling chemical restored the sequestration of mitochondria into autophagosomes in Bnip3L−/− erythrocytes (Fig. 1C). Our study indicates that Bnip3L-dependent loss of Δ Ψ m is important for targeting the mitochondria into autophagosomes for degradation during erythroid maturation, and interference with this function impairs terminal erythroid maturation and results in hemolysis. These data underlie the importance of pro-apoptotic mechanisms for productive terminal erythropoiesis through autophagy.

Figure Figure
Emodin Induces Leukemic HL-60 Cells Apoptosis by Inhibiting PI3K/Akt Signal Pathway.
Authors: Hu Jian Da , Zheng He-yong , Zheng Zhi-hong , Huang Lv-ye , Lv Lian-huang , Chen Ying-yu
Published: October 2019

Emodin (1.3.8-trihydroxy-6-methylanthraquinone), a traditional Chinese herbal medicine, which is a natural anthraquinone compound from the root and rhizome of Rheum palmatum, has been shown to display a number of biological activities such as antiviral, antimicrobial, immunosuppressive, anti-inflammatory, and anticancer. Recent studies showed that it can inhibit proliferation of multiple cancer cells such as breast, hepatocellular, and lung cancer cells in vitro. The PI3K signal transduction pathway has been investigated extensively for its role in oncogenic transformation and in the prevention of apoptosis. The aim of the present study was to investigate the effects of Emodin on inhibiting cell growth and inducing apoptosis in human myeloid leukemia cell line HL-60, and to examine how emodin affects the PI3K/AKT signalling pathway, leading to cell apoptosis. HL-60 cells were exposed to various dosages of Emodin. MTT assay was used to detect HL-60 cell proliferation. Distribution of HL-60 cells in cell cycle was analyzed by flow cytometry and cell apoptosis was observed by MitoCapture apoptosis detection. The protein expressions of Akt signal pathway were detected by Western-Blot. The results showed that Emodin remarkably inhibited the cell proliferation, with 20μmol/L of the IC50 value for 48h. Apoptosis in HL-60 cells could be efficiently induced by Emodin in a dose dependent manner and cells were arrested at G0/G1. The expressions of Akt,p-Akt,IκB-α,p-IκB-α,p65,p-p65,mTOR and p-mTOR in PI3K/Akt signal pathway were downregulated after Emodin treatment. It could be concluded that Emodin can efficiently induce growth inhibition and apoptosis in HL-60 cells .Akt signal pathway may be involved in this process.
High INDO (Indoleamine 2,3-Dioxygenase) mRNA Level in Blasts of Acute Myeloid Leukemic Patients Predicts Poor Clinical Outcome.
Authors: Chamuleau Martine E.D. , Van de Loosdrecht Arjan A. , Hess Corine J. , Janssen Jeroen J.W.M. , Zevenbergen Adri , Delwel Ruud , Valk Peter J.M. , Lowenberg Bob , Ossenkoppele Gert J.
Published: October 2019

Indoleamine 2,3-dioxygenase (IDO) degrades the amino acid tryptophan which is essential for T cells. Tryptophan depletion causes T cell cycle arrest and solid tumors that express high levels of IDO can create immune suppression. Recently, blasts of acute myeloid leukemic (AML) patients were shown to express the IDO protein. We now determined INDO (the encoding gene for IDO) mRNA expression in myeloid leukemic blasts of 285 AML patients by gene expression profiling. Results were validated by quantitative PCR (qPCR) analysis in blasts of an independent cohort of 71 patients. Correlation of INDO expression to relevant known prognostic factors and survival identified high INDO expression as a strong negative independent predicting variable for overall and relapse free survival of AML patients. Inhibition of IDO may result in breaking tumor induced immune tolerance and offers new therapeutic options for AML patients.

Figure Figure
Activating KIR (B Haplotype) in Unrelated Donors Improves Disease-Free Survival after HLA-Matched and Mismatched Allogeneic Transplantation for Acute Myeloid Leukemia: Implications for Donor Selection Using KIR Genotyping.
Authors: Cooley Sarah , Miller Jeffrey S. , Trachtenberg Elizabeth , Bergemann Tracy , Saeteurn Koy , Klein John , Guethlein Lisbeth A. , Marsh Steven G.E. , Geraghty Daniel , Parham Peter , Weisdorf Daniel
Published: October 2019

The importance of killer immunoglobulin-like receptors (KIR) in determining clinical outcome after hematopoietic cell transplantation (HCT) remains controversial. We analyzed 428 unrelated, T-cell replete transplants for acute myeloid leukemia (AML) facilitated by the National Marrow Donor Program between 1988 and 2003. Fifty seven KIR-ligand mismatched (thus HLA mismatched; KIR-Lmm/HLAmm) transplants were selected as cases. By matching on 15 clinical and demographic variables, KIR ligand matched, HLA mismatched (KIR-Lm/HLAmm; n = 162) and 10/10 HLA matched (HLAm; n = 209) comparison groups were selected. Using a validated single nucleotide polymorphism (SNP)-based MALDI-TOF assay, DNA samples were typed for 16 individual KIR genes which were used to estimate KIR haplotypes. About 1/3 of donors and recipients were A/A (lacking any activating KIR except 2DS4) and 2/3 possessed at least one B haplotype (B/X). As expected, 3 year survival was better in HLAm vs. HLAmm KIR-Lm or KIR-Lmm HCT (32%; p = .07 and 0.0004 respectively). However, survival was similar in both HLAmm groups (KIR-Lm 27% vs. KIR-Lmm 17%; p = .11). Importantly, transplants from donors with an activating B KIR haplotype had significantly better survival. Using a Cox regression that stratifies the baseline hazard according to sets of patients matched on important clinical variables, transplants using B KIR haplotype donors had a 42% reduced risk of mortality [B/X (vs. A/A) KIR haplotype donors (RR .58, 95% CI .44–.78; p = .0002]. Similarly, for disease-free survival (DFS), the risk of relapse and/or death decreased 43% [B/X (vs. A/A) RR .57, 95% CI .43–.76; p = .0001]. No differences were seen in risks of acute GVHD. The significant improvements in overall survival and DFS conferred by donor B KIR haplotype were seen in both the HLAm and KIR-Lm/HLAmm groups, but not in the KIR-Lmm/HLAmm group. The recipient KIR haplotype had no effect on outcomes, and the benefit of donor B KIR haplotype was equal regardless of recipient KIR haplotype (figure). Individual KIR genes were also analyzed and in Cox regression only donor 2DL2 and 2DS2 were favorable. As these genes are in strong linkage disequilibrium, the effects were similar (donor 2DL2 RR of death .70, 95% CI .54–.91; p = .008 and 2DS2 RR .66, 95% CI .50–.86; p = .002). 2DS2 defines a B KIR haplotype, confounding interpretation of these single gene effects. In summary, data from this analysis of the effect of KIR genes in unrelated T-replete HCT for AML suggest that using B KIR haplotype donors improves survival and DFS. Therefore, KIR genotyping could be incorporated into unrelated donor selection algorithms to identify B haplotype donors for patients with AML.

Figure Figure
A Study of the Protein Phosphatase 2A Involved in the Induction of NB4 and MR2 Cell Differentiation by ATRA.
Authors: Ouyang Jian , Xu Xihui , Wu De Pei
Published: October 2019

Objective: To study whether protein phosphatases type 2A (PP2A) plays an important role in signal transduction of differentiation of NB4 induced by all-trans retinoic acid, furthermore, to evaluate whether PP2A plays an important role in MR2 resistance to ATRA.

Methods: ATRA, Okadaic acid (OKA) and both chemicals were incubated with NB4 and MR2 cells respectively. Wright’ staining and NBT reduction test were employed to evaluate the changes of cell phenotype. The expression of CD11b was measured by flow cytometer, and the proliferation of cells was measured by MTT method. At last the activity of PP2A was evaluated by Serine/Threonine phosphatase assay system.

Results: Results showed ATRA induced differentiation of NB4 significantly, but no effect on MR2 cell through Wright’ staining and NBT reduction test. OKA, the specific inhibitor of PP2A, could augment the differentiation induced by ATRA, and it combined with ATRA showed the slight induction of differentiation of MR2 cell. MTT showed the inhibitory rate of proliferation of ATRA, OKA and ATRA combined with OKA to NB4 were 25.1%, 10.1% and 38.4% respectively, and to MR2 were 4.8%, 7.2% and 19.3%. When ATRA, and ATRA combined with OKA were incubated with NB4 and MR2 respectively, the expression of CD11b on the surface of NB4 cell were 43.1% and 65.4% and on the surface of MR2 were from 0 to 9.3%. When OKA combined with ATRA were incubated with MR2, MR2 have showed the decreased activity of PPA. Conclusion PP2A plays an important role in the mechanism of differentiation of NB4 induced by ATRA and of resistance of MR2 to ATRA.
Downregulation of Peroxiredoxin III Contributes to Arsenic Trioxide-Induced Apoptosis in APL.
Authors: Vivas-Mejia Pablo , Ozpolat Bulent , Chen Xian , Lopez-Berestein Gabriel
Published: October 2019

Arsenic trioxide (ATO) induces differentiation and apoptosis in acute promyelocytic leukemia (APL). Compared with other leukemia cells, the APL-derived NB4 cells are particularly sensitive to ATO-induced apoptosis. Several reports indicate that in NB4 cells, apoptosis occurs in part by a mechanism that involves the inhibition of glutathione peroxidase, one of the enzymes that regulate the levels of H2O2 in the mitochondria, and that is present only at low levels in NB4 cells. Peroxirredoxin III (Prx III) is a mitochondria-specific H2O2-scavenger member of the thioredoxin-dependent peroxidases. NB4 cells express high levels of Prx III; however the role of Prx III in ATO-induced apoptosis in NB4 cells has not been investigated. We studied here whether Prx III is regulated during ATO-induced apoptosis in NB4 cells and whether the depletion of Prx III further sensitized these cells to ATO-induced apoptosis. The protein and mRNA levels of Prx III were decreased during ATO-induced apoptosis of NB4 cells. The down-regulation of Prx III occurred prior to the accumulation of reactive oxygen species, reduction in the mitochondrial membrane potential and apoptosis. Depletion of Prx III enhanced the ATO-induced mitochondrial damage, and also promoted cytochrome-c release, and caspase-3 and caspase-9 activation. Prx III is downregulated early during ATO-induced apoptosis, facilitating in this way the mitochondria-depending apoptotic events triggered by the accumulation of H2O2.
Traumatic Lumbar Puncture (TLP) Is Not Associated with Worse Outcome in Adults with Acute Lymphocytic Leukemia (ALL) or Burkitt-Type Leukemia/Lymphoma (BLL) Receiving Hyper-CVAD Regimens.
Authors: Atallah Ehab , O’Brien Susan , Kaled Susan , Lynn Alice , Cortes Jorge , Kantarjian Hagop , Thomas Deborah A.
Published: October 2019

Background: TLP has been associated with worse outcome in pediatric ALL patients (pts). However, this has not been well studied in adults with ALL or BLL. Our aim was to investigate the prognostic significance of CNS status at presentation in adult ALL pts undergoing frontline therapy with hyper-CVAD (fractionated cyclophosphamide, vincristine, doxorubicin, and dexamethasone alternating with methotrexate and cytarabine) or variations incorporating rituximab and/or imatinib. Pts were stratified to receive standard intrathecal treatments (IT) with methotrexate alternating with cytarabine based on CNS risk assessment (proliferative index ≥ 14% or serum lactate dehydrogenase ≥ 1400) [Cortes et al, Blood 86:2091, 1995]. Low risk pts received 6 IT, high or indeterminate risk 8 IT, and BLL 16 IT.

Methods and Results: Cerebrospinal fluid was assessed after IT approximately day 2 of induction therapy. CNS status at presentation was defined as: CNS 1 (not traumatic; < 10 RBCs/μL without leukemic blasts after cytocentrifugation); CNS 2 (not traumatic; < 5 WBCs/μL with leukemic blasts); CNS 3 (not traumatic; ≥ 5 WBCs/μL with leukemic blasts), TLP+ (traumatic, > 10 RBCs/μL with leukemic blasts). TLP- (traumatic without leukemic blasts). Two hundred eighty two pts were reviewed up to January 2007; the median age was 42 years (range, 15–84). Diagnosis was ALL in 229 (81%) and BLL in 53 (19%). In the ALL subset, 48 pts (17%) were Philadelphia positive. Median WBC and platelet count were 3.2 x 109/L and 65 x 109/L, respectively. At the time of first LP, 119 (42%) pts had circulating (PB) blasts. CNS risk was high in 34% (plus 17% with BLL), indeterminate in 20%, and low in 21%. The remainder (7%) had definitive CNS disease and were not classified. PB blasts and CNS risk did not appear to affect the CNS relapse rate. By multivariate analysis, age, thrombocytopenia, leukocytosis and presence of t(4;11) were associated with worse survival in this selected group, while CNS status at presentation was not (Table 1).

CNS status and risk of CNS relapse CNS status at 1st LP No. (%) %CNS Relapse CNS1 159 (56) 5 CNS2 13 (5) 15 CNS3 7 (3) 14 TLP+ 25 (9) 4 TLP− 78 (27) 4

Conclusion: CNS status at first LP does not appear to affect prognosis of adults with de novo ALL or BLL receiving frontline therapy with hyper-CVAD based regimens. TLP or the presence of PB blasts at the time of first LP does not appear to increase the risk of CNS relapse.
Impact on Quality of Life of Postconsolidation Immunotherapy with Histamine Dihydrochloride and Interleukin-2 in Acute Myelogenous Leukemia.
Authors: Wallhult Elisabeth , Whisnant John , Rowe Jacob M. , Szer Jeff , Bhagwat Dileep , Hellstrand Kristoffer , Nilsson Bo I. , Brune Mats L.
Published: October 2019

PURPOSE: To assess the impact on Quality of Life (QoL) of postconsolidation immunotherapy with histamine dihydrochloride (HDC) in combination with interleukin-2 (IL-2) in acute myelogenous leukemia (AML).

PATIENTS & METHODS: A multi-national randomized open-label phase 3 trial recruited 320 AML patients between 1998 and 2000. Patients were enrolled after termination of consolidation therapy, stratified by country and remission status (first or subsequent remission), and randomized to HDC/IL-2 treatment or no treatment (standard-of-care, control). The treatment was self-administered at home and included ten 3-week courses of HDC (0.5 mg sc bid; EpiCept Corporation) plus low-dose recombinant IL-2 (aldesleukin, 18,000 U/kg sc bid; Chiron Corp). Cycles 1–3 comprised 3 wks of treatment followed by 3 wks of rest, whereas in cycles 4–10 the rest periods were 6 wks. As reported previously (Blood2006; 108:88), the trial met the primary endpoint of prolonged leukemia-free survival (LFS) for all patients randomized (P=0.008). Assessment of QoL, a secondary trial endpoint, was performed using the validated EORTC-C30(v2) instrument at 8 pre-defined study visits: baseline; pre- and post-cycles 3, 5, and 8; and at 18 months. Data were analyzed as specified in the EORTC scoring manual for 9 scales (1 global, 5 functional, 3 symptom scales) and 6 single-symptom items. Comparisons were made between and within groups at

baseline vs. each visit, pre- vs. post-treatment, and baseline vs. end of treatment (18 months).

RESULTS: At least one questionnaire was completed by 285 patients (89%). Patients in both study arms increased or maintained QoL status from baseline to last evaluation with respect to global health status, physical, cognitive and role functioning, and financial issues, as well as symptoms of fatigue, nausea/vomiting, pain, diarrhea, and dyspnea. Comparisons of pre- vs. post-treatment QoL assessments showed transient increases in fatigue (P=0.008), nausea/vomiting (P=0.006), and appetite loss (P=0.005) in the treatment arm.

CONCLUSION: Post-consolidation immunotherapy with HDC/IL-2 self-administered at home by AML patients was associated with transient gastrointestinal symptoms and fatigue, but did not prevent recovery from induction/consolidation chemotherapy.
Intensity of FDG Uptake on PET Scan Varies by Histologic Subtype of Hodgkin Lymphoma.
Authors: Holalkere Nagaraj , Hochberg Ephraim P. , Takvorian Ronald , Blake Michael , Toomey Christiana , Michaelson James , Rahemtullah Aliyah , Abramson Jeremy S.
Published: October 2019

FDG-PET scanning has demonstrated utility in the staging and prognostication of Hodgkin lymphoma, but PET characteristics of discrete histologic subsets of Hodgkin lymphoma have never been reported. Further, while studies in this disease have focused on classical Hodgkin lymphoma, the PET features of nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) have not been addressed to date. NLPHL represents less than 5% of all cases of Hodgkin lymphoma and has distinct clinicopathologic features, natural history, and treatment from classical subtypes. Diagnosis may be difficult as it exists in a “gray zone” between Hodgkin and non-Hodgkin lymphomas (NHL), and shares immunophenotypic and histologic features with the T-cell rich variant of diffuse large B-cell lymphoma. Clinically, NLPHL follows a natural history more akin to indolent B-cell NHLs than to classical Hodgkin lymphomas. We retrospectively identified 7 cases of NLPHL diagnosed at our institution since 2003 with available PET staging data. Consistent with prior series of this disease, the majority of patients were male (6:1) and the median age was 32 years. All 7 patients presented with limited stage disease (3 stage I, 4 stage II) and without “B” symptoms. All 7 cases demonstrated increased FDG avidity on PET scanning. The mean Standard Uptake Value (SUV) max was 6.1 (range 1.1–8.8), and mean SUV mean was 5.6 (range 1.0–8.3). We compared these results to recently diagnosed patients with nodular sclerosis and mixed cellularity subtypes of classical Hodgkin lymphoma. Among 13 patients with nodular sclerosis Hodgkin lymphoma (NSHL), the mean SUV max was 10.1 (range 4.8–15.8), while among the 7 patients with mixed cellularity histology (MCHL), the mean SUV max was 5.6 (range 4.0–9.8). Patients with NLPHL had significantly decreased SUV max compared to NSHL patients (p=0.022), but were similar to patients with MCHL (Figure 1). Mixed cellularity histology similarly demonstrated significantly decreased SUV max compared to patients with nodular sclerosing disease (p=0.005). PET intensity therefore highlights differences between classical subtypes of Hodgkin lymphoma, as well as between classical and non-classical variants. When compared to published PET SUV data in aggressive B-cell lymphomas, the maximal SUVs we observe in NLPHL fall well below those reported in aggressive B-cell lymphomas, which may aid in the diagnosis of these occasionally similar appearing neoplasms. We conclude that PET scans identify differences in FDG avidity across classical and non-classical Hodgkin lymphoma subtypes, likely reflecting underlying biological differences. We also demonstrate for the first time that despite its indolent behavior, NLPHL is a uniformly FDG-avid neoplasm, and so PET scans may have a role in the staging and surveillance of this uncommon disease variant.

Figure Figure
Velcade®, Thalidomide, Dexamethasone (VTD) Induction Therapy Followed by Melphalan, Prednisone, Thalidomide (MPT) Maintenance as a First Line Treatment for the Patients (pts) with Multiple Myeloma (MM) Who Are Non-Transplant Candidates: Early Analysis from the Korean Multiple Myeloma Working Party.
Authors: Eom HyeonSeok , Kim Yeo-Kyeoung , Chung Jooseop , Kim Kihyun , Kim HyoJeong , Jin JongYul , Do Young-Rok , Oh Sukjoong , Kim HoYoung , Suh Cheolwon , Seong Chu-Myong , Kim Chul Soo , Lee Dong Soon , Lee Jae Hoon
Published: October 2019

VTD and MPT chemotherapies have been known to be active regimens in pts with MM. The objective of this study is to examine response and toxicities and to estimate survival of pts with VTD followed by MPT, who are non-transplant candidates with previously untreated MM. Total of 29 pts were enrolled from March, 2006 through August, 2007 and this study is still ongoing. 13 pts were men and 16 pts were women. The median age was 67 years (range, 61–75 years) and median follow up was 5 months (range, 1–16 months). Here 23 pts who completed at least first two cycles of VTD were analyzed. Pts received bortezomib (Velcade®) 1.3 mg/m2 on days 1, 4, 8, 11, thalidomide 100 mg daily, dexamethasone 40 mg on days 1–4 every 3 weeks for a maximum of 6 cycles of treatment, and thereafter melphalan 4 mg/m2 on days 1–7, prednisone 40 mg/m2 days 1–7, thalidomide 100 mg daily every 4 weeks for a maximum of 12 cycles. In these 23 pts, 92% of them showed responses (18% CR, 4% nCR, and 70% PR). 17 pts completed 4 cycles of VTD and all of them showed 100% response rates (CR 35%, nCR 18%, PR 47%). 13 out of 14 who completed 6 cycles of VTD showed responses (CR 50%, nCR 14%, PR 29%, PD 7%). It is too early to see whether improved response rate translates into improved overall survival (OS) and progression free survival (PFS). The median OS and PFS have not been reached yet. 11 pts (37%) stopped protocol therapy because of consent withdrawal (2 pts), death (4 pts), disease progression (2 pts) and severe adverse reaction (3 pts). The causes of death were infection-related in 2 pts who had been in remission. Other 2 pts were related to disease progression. Although peripheral neuropathy affected all of pts, only 20% of the pts were grade 3. The most common side effects of the chemotherapies greater than grade 3 were pneumonia (24%), asthenia (12%), diarrhea (16%), nausea (4%), thrombocytopenia (16%), neutropenia (12%) and anemia (12%). Although VTD followed by MPT chemotherapy in pts with previously untreated MM, who are non-transplant candidates showed high response rates with manageable toxicities, they showed high withdrawal rates from the study which attributed partly to the characteristics of the pts at baseline who were non-transplant candidates because of old age and morbidities, and a few major neuropathies. Follow up data will be presented.