bluuebunny/crossref_metadata_2025 huggingface dataset:
IL-6 Enhances C-myc Translation in Myeloma Cells Via Activation of the Myc Internal Ribosomal Entry Site (IRES) and an IRES-Binding Trans Factor, hnRNP A1.
Authors: Shi Yijiang , Gera Joseph , Lichtenstein Alan
Published: October 2019

Dysregulated c-myc expression in multiple myeloma (MM) can be due to a translocation with juxtaposition of the myc gene to the Ig enhancer resulting in constitutive transcription. However, myc protein levels can also be increased in MM cells when it is due to heightened translation. Activated translation can be mediated via an mTOR-mediated, cap-dependent mechanism, thus named because the translation initiation complex utilizes the cap structure on mRNA to recruit the ribosomal subunit. However, an alternative mechanism is cap-independent where the ribosomal subunit binds a transcript via an IRES in the RNA’s 5′UTR. IRES function is regulated by IRES trans activating factors (ITAFs) which bind to the 5′UTR. We studied myc expression in ANBL-6 wild type and activated K-RAS-transfected MM cells which proliferatively respond to IL-6 but do not contain myc translocations. IL-6 increased myc protein expression in both cell lines in a concentration and time-dependent fashion but RNA expression was only enhanced in the wild type cells. The increased protein expression in K-RAS mutant cells was unaffected by rapamycin, indicating independence from mTOR and cap-dependent mechanisms. Using an IRES-dependent reporter vector, reporter expression was shown to be increased by IL-6, confirming IL-6 activates myc IRES activity. This was also demonstrated in the IL-6 autocrine U266 MM cell line where myc IRES-dependent reporter expression was ablated following exposure to anti-IL-6 antibodies. In addition, primary MM cells treated with IL-6 also demonstrated an increase in myc IRES activity. Since a yeast three-hybrid screen for proteins specifically binding to the myc IRES identified several clones encoding hnRNP A1, we evaluated hnRNP A1 as a mediator of myc IRES activity. IL-6 increased the cytoplasmic localization of hnRNP A1 in ANBL-6 MM cells. Furthermore, in MM cells where hnRNP A1 expression was knocked down, myc IRES activity was greatly diminished and there was no enhancement with addition of IL-6. These results indicate that IL-6 can enhance c-myc translation in MM cells via activation of the myc IRES and that hnRNP A1 is a critical myc ITAF involved in the IL-6 response. hnRNP A1 may serve as a new therapeutic target in myeloma.
TH17 Pathway and Associated Pro-Inflammatory Cytokines Promote Immune Dysfunction in Myeloma.
Authors: Prabhala Rao , Pelluru Dheeraj , Neri Paola , Fulciniti Mariateresa , Driscoll James J. , Weihua Song , Blotta Simona , Tai Yu-Tzu , Patel Hiren , Efebera Yvonne A. , Hideshima Teru , Chauhan Dhaminder , Daley John F. , Anderson Kenneth C. , Munshi Nikhil C.
Published: October 2019

Multiple myeloma (MM) is associated with significant immune dysfunction. Although various mechanisms mediating immune dysregulation in MM have been studied, its molecular and cellular basis is ill defined. IL-6, TGF-β and IL-1β have been implicated in this process, but their mechanism of effects on immune function have not been studied in MM. Together, IL-6 and TGF-β enhance the generation of TH17 cells, important in the development of immunity and auto-immunity. Additionally, TH17 cells are differentiated by number of inflammatory cytokines including, IL-21, IL-22, IL-23, and IL-27. Therefore, we evaluated the immune dysfunction and the role of TH17 cells and associated pro-inflammatory cytokines in myeloma. We have previously characterized that the production of TH1 mediated cytokines including IFN-γ following anti-CD3-mediated activation is significantly lower in myeloma PBMC compared to normal PBMC. We hypothesize that this may be regulated via skewing the immune system towards TH17 pathway. We observed that TH17 cells, measured by intra-cellular flow cytometry, are significantly increased in number in myeloma (16.9%) and MGUS (6.2%) compared to normal (3.3%). Furthermore, we analysed supporting pro-inflammatory cytokine network for the generation of TH17 cells in myeloma, which may be responsible for the observed TH17 skewing of T cell subsets. Sera from MGUS (n=12) and myeloma (n=17) patients were evaluated for the presence of these pro-inflammatory cytokines compared with normal sera (n=6) using ELISA. We observed significant increase in serum IL-21, IL-22 and IL-23 in MGUS (373 pg/ml, 14 pg/ml and 147 pg/ml respectively; p<0.05) and myeloma (296 pg/ml, 12 pg/ml and 215 pg/ml respectively; p<0.05) compared with normal (63 pg/ml, 1.5 pg/ml and 39 pg/ml respectively). In addition, we also observed that the myeloma PBMC stimulated in the presence of IL-6 and TGF-β, both of the cytokines present at a high level in myeloma, induced significant IL-23 production compared with normal. Importantly, IL-23 levels were 10 fold higher in myeloma BM samples compared with matching blood samples. These results indicate that the cytokines from myeloma BM microenvironment may be responsible for the observed T cell subset abnormality by favouring TH17 cells via IL-23/IL-21 production. These cytokines thus may be targets to modulate immune responses in myeloma to enhance immune function and devise effective vaccination strategies in the future.
Mycosis Fungoide: Immunochemistry Analysis of Lymphoid and Microenvironment Cells by Macrotissue Array.
Authors: del Agua Celia , Rubio-Martinez Araceli , Filipo Francesc , Piris Miguel A. , Giraldo Pilar
Published: October 2019

Aims: Cutaneous lymphomas (CL) represent a unique group of lymphomas and are the second most frequent extranodal lymphomas. CL probably is the result of a multifactor and multistep process. Firstly an inflammatory reactive process is developing secondary to various chronic stimuli that may be genetic, environmental, infectious and immunologic. The consequences are the negative effects in cell regulation and deregulation of oncogenes and/or suppressor genes later promotes transition from pre-neoplastic conditions to neoplasia. Detailed molecular expression analysis of cutaneous T-cell lymphoma is not available. Some oncogenic alterations have been demonstrated, such as functional inactivation of the Fas receptor, constitutive activity of STAT 3, or the inactivation of the p16 gene via deletion or promoter hypermethylation.

Objective: To study the expression of p16, c-myc, Ki67, bcl-2, CD1a, CD123, TCL1, CD68, STAT 3, STAT 4 and MAL1 in an homogeneous group of. CL diagnosed in a general Hospital in order to know more about the expression of these markers in this neoplastic process.

Design: retrospective, analytical study in 30 CL diagnosed consecutively as Mycosis Fungoide (14 early and 16 advanced stages) since January 2005 to December 2006. A macrotissue array technique by incubation with monoclonal antibodies and immunohistochemistry protocol to visualize by avidin-biotin peroxidases was applied in all paraffin histological samples. A semi quantitative method was applied in order to categorize the positive cells.

Results: In 28 samples (92%) we have observed p16 positive over expression, the two negative samples corresponding to early affectation. In 14 samples from early stages (48%) c-myc was negative. In 29 samples (96%) CD1a was positive in dermal and epidermal layer. CD123 (interleukine 3 receptor) was negative in 16 samples (52%) and TCL1 was positive in 12 cases (39%) in small cells with oval and cleaved nucleus and scarce cytoplasm. STAT3 and STAT4 were positive in 29 cases (97%), Over expression of MAL1 (48%) was observed in advanced patients with aggressive CL.

Conclusions: In our study an over expression of p16 is observed in the majority of cases and high c-myc expression in advanced stages. Probably dendritic plasmacytic cells are involved in the pathogenesis of skin lymphoproliferative disorders with cutaneous T cell infiltration. MAL1 could be a predictor of aggressive CL but it is necessary more studies and more cases in order to confirm it.
Primary Non-Hodgkin’s Lymphoma of the Lung.
Authors: Fong Thomas , Lossos Izidore , Morgensztern Daniel
Published: October 2019

Background: Unlike the relatively common secondary involvement of the lung in patients with widespread disease, primary lung lymphoma (PLL) is a rare disease with a few reports in the literature and information based mainly from small series of patients. We conducted a study in a large population database to improve the current knowledge of this malignancy.

Methods: The SEER database was searched for primary non-Hodgkin’s lymphoma (NHL) of the lung in patients aged 20 or older and diagnosed between 1988 and 2003. Entry criteria included lung malignancies with codes for histology 9670–9699 (mature B-cell lymphomas), 9700–9719 (mature T and NK-cell lymphomas), 9720–9729 (precursor cell lymphoblastic lymphomas) and NHL at primary sites labeled C34.0 (main bronchus), C34.1 (upper lobe), C34.2 (middle lobe), C34.3 (lower lobe), C34.8 (overlapping), and C34.9 (lung NOS). All primary lung NHLs (also including histology codes 9590–9599, malignant lymphoma, NOS or diffuse) were used in the calculation as a percentage of total NHLs and primary lung tumors, but only patients with known histology were used for further analysis.

Results: There were 1,652 cases of PLL, which accounted for 5.2% of all extranodal NHL and 1.7% of all NHL. PLL represented 0.4% of all primary lung cancers, but 51% of non-carcinoma histologies. Among the PLL cases, 355 did not have histological definition, leaving 1,297 eligible for further analysis. The median age at diagnosis was 65 years, and the disease was slightly more common in males (53.2% male; male/female ratio of 1.14). Most of the patients were Caucasians (85.6%), and the most common histologies were diffuse large B-cell (DLBCL) (51%), marginal zone B-cell lymphoma (MZL) (25.2%), small lymphocytic lymphoma (SLL) (9.2%), and follicular lymphoma (FL) (5.3%). Stage distribution was as follows: stage IE - 45.2%, stage IIE - 18.1%, stage IIIE/IIIES - 2.7%, and stage IV - 34.0%. Most patients presented with early-stage (IE/IIE) disease (63.3%). Surgery and radiation were the initial therapy in 36.2% and 13.1% of the patients respectively. Median overall survival was 58 months, and 5-year survival was 48.5%. The 5-year survival rates for patients with early stage disease (IE and IIE) according to histology were as follows: DLBCL - 36%, MZL - 78%, SLL - 71%, and FL - 81%.

Conclusions: PLL is a rare disease with limited information from the literature. Although it accounts for a small fraction of both NHLs and primary lung tumors, it is the most common pulmonary malignancy of non-carcinoma histology. It occurs most commonly in elderly patients, with a slight male predominance. Nearly two-thirds of the patients present with early-stage disease, and approximately half of the patients are alive at 5 years.
First Successful Allogeneic Hematopoietic Stem Cell Transplantation in Children with Chronic Myeloid Leukemia, Preceded by Imatinib Prior Transplantation.
Authors: Wu Kang-Hsi , Weng Te-Fu , Chen Ming-Chieh , Peng Ching-Tien
Published: October 2019

Objectives: Although very rare in children, chronic myeloid leukemia (CML) can be cured by hematopoietic stem cell transplantation (HSCT). However, the 5-year survival probability following allogeneic HSCT is only about 50–60% in childhood CML. With imatinib successful in CML patients, the impact of imatinib treatment on subsequent allogeneic hematopoietic stem cell transplantation (HSCT) remains unclear. Imatinib therapy prior to HSCT has not been reported in children. Herein we describe imatinib induction preceding HSCT in three children newly diagnosed.

Methods: Imatinib was administered as induction in three boys, aged 11, 14, 15 years, newly diagnosed in the chronic phase. Between induction and cytogenetic remission, two patients received HSCT from matched unrelated donors (MUD) and one received HSCT from matched sibling donor. Busulphan and cyclophosphamide were utilized as a conditioning regimen. Cyclosphorin A and methotrexate were both used as graft versus host disease (GVHD) prophylaxis. Anti-thymocyte globulin was added for MUD cases.

Results: Successful engraftment and neither severe liver toxicity nor acute GVHD were encountered. All three cases show cytogenetic remission, with chronic GVHD well controlled in one. We monitored each patient’s status for minimal residual disease by real-time quantitative polymerase chain reaction. Absence of detectable minimal residual disease was found in two patients at 28 and 63 months after HSCT. At present, all of these three patients have good quality of life.

Conclusions: As first-line therapy, imatinib appears useful for providing a bridge to HSCT in children with CML. Further study is warranted to determine impact of imatinib prior transplantation on child CML.
Dasatinib Can Be Administered Orally with or without a Meal.
Authors: Kaul Sanjeev , Wu Chiyuan , Mayfield Shelley , Manning James , Blackwood-Chirchir Anne
Published: October 2019

Background: Food can change the bioavailability of a drug, which can have clinically significant consequences. This study was conducted to investigate the effect of food on the oral bioavailability of dasatinib (SPRYCEL®) in healthy adult subjects.

Methods: Fifty-four healthy adult subjects received a single dose of dasatinib 100 mg dose, as 2 x 50 mg film-coated tablets after an overnight fast and within 10 minutes after the ingestion of a low-fat meal (315 kcal [20% fat, 68% carbohydrates, and 12% protein]) and a high-fat meal (985 kcal [52% fat, 34% carbohydrates, and 14% protein]) in a randomly assigned sequence. Individual treatments were separated by at least a 7-day washout period. Serial blood samples were collected for 24 hours after each treatment to determine dasatinib plasma concentrations using a validated liquid chromatography/tandem mass spectrometric method. Dasatinib pharmacokinetic (PK) parameters were determined using a non-compartmental method. Safety was monitored throughout the study.

Results: Of the 54 healthy adult subjects (85% male, 61% Caucasian, mean age 32 y, and weight 80 kg), 48 completed the study. There were no serious adverse events. Adverse events and laboratory abnormalities were, in general, typical of those seen with dasatinib administration. PK results are summarized in the table below.

Conclusions: Compared to the fasted state, a low-fat meal decreased Cmax and AUC of dasatinib by 21%; a high-fat meal decreased Cmax by 24% and increased AUC by 14%. These results are not expected to be of clinical relevance and, therefore, dasatinib may be taken without regard to meals. The drug was generally safe and well-tolerated when administered in the fed or fasted state.

Statistical Analysis of PK Parameters for Dasatinib Treatment PK Parameter Geometric Mean Ratios (95% Confidence Intervals) Fed versus Fasted Low-Fat Meal Cmax 1.216 (1.047, 1.413) AUC 1.212 (1.100, 1.336) High-Fat Meal Cmax 0.758 (0.651, 0.882) AUC 1.140 (1.034, 1.257)
Clinical Features and Prognosis of MDS Patients with Chromosome 5 Abnormalities Other Than ‘5q-Syndrome’: Results of Multi-Center Analysis in Korea.
Authors: Cheong June-Won , Kim Inho , Yoon Sung-Soo , Lee Dong Soon , Jung Chul Won , Ahn Jin Seok , Kim Kihyun , Kim Sun Hee , Seo Eul-Ju , Park Chan-Jeoung , Chi Hyun-Sook , Kim Sung-Hyun , Kwon Hyuk-Chan , Kim Hyo-Jin , Kim Hyeoung Joon , Ahn Jae Sook , Kook Hoon , Shin Ho-Jin , Chung Joo Seop , Lee Mark Hong , Yoon Hwi-Joong , Bae Sung Hwa , Joo Yong-Dong , Hong Dae Sik , Bang Soo-Mee , Kim Chul Soo , Jo Deog-Yeon , Zang Dae Young , Hwang Doh Yu , Kim Jin Seok , Min Yoo-Hong
Published: October 2019

Introduction: The myelodysplastic syndrome (MDS) is frequently associated with various chromosomal abnormalities. ‘5q− syndrome’ is low-risk MDS known as good responder of lenalidomide recently. However, the patients with other abnormalities in chromosome 5 showed quite different clinical features from those with ‘5q− syndrome’. The aim of this study was a retrospective evaluation for Korean MDS patients with abnormalities in chromosome 5 other than ‘5q− syndrome’.

Materials and Methods: Among 456 patients with MDS diagnosed at 16 hospitals in Korea between 1996 and 2006, 370 with available cytogenetic data entered the study. Univariate and multivariate analysis were performed.

Results: Ninety three patients (25.1%) showed abnormalities in chromosome 5 and the ‘5q− syndorme’ was only 10 patients (2.7%). Among the rest, 39 patients (10.5%) had various abnormalities other than 5q deletion such as translocation or 5 monosomy, 38 (10.3%) had complex abnormalities with 5q−, and 2 had mosaic pattern with normal chromosome. Four patients had isolated 5q− but blasts in marrow were over 5%. The deletion of 5q was interstitial but with a predominance for 5q13-33 deletions (34.8%). MDS patients with chromosome 5 abnormalities other than ‘5q− syndrome’ didn’t share the clinical features with ‘5q− syndrome’. There was no leukemic transformation in ‘5q− syndrome’ group, but 18 (21.7%) with other abnormalities in chromosome 5 finally transformed to acute leukemia. Five year overall survival was significantly inferior in non-’5q− syndrome’ patients than ‘5q− syndrome’ (14.3% vs. 79.6%, P=0.0115).

Conclusions: Patients with isolated 5q− and excess blast (>5%), other abnormalities than isolated 5q−, or mosaic chromosome with isolated 5q− and normal chromosome didn’t share the clinical features such as lower rate of leukemic transformation and long survival.
Establishment of a Metastasizing Human Cell Line Based Multiple Myeloma Model in Immunodeficient Mice.
Authors: Schueler Julia B. , Wider Dagmar , Wagner Martin , Fiebig Heiner , Engelhardt Monika
Published: October 2019

Multiple myeloma (MM) accounts for 10% of all hematological malignancies, leads to devastating bone destruction, bone pain, renal failure, and hematopoietic insufficiency. In order to allow a better understanding of MM, the establishment of functional and reproducible in vivo models is pursued worldwide. Of available models, xenograft models in immunodeficient mice (IDM) reproduce the clinical situation best. Using the different MM cell lines (MMCLs) L363 and RPMI8226, we tested their ability to engraft in IDM under different conditions. As the supportive effect of bone marrow (BM) stroma has been suggested to be vital, we injected L363 or RPMI8226 into a freshly prepared mouse tibia which was implanted subcutaneously (sc) into IDM. In a second approach, we injected MMCLs directly into the tibia of the recipient mouse. Furthermore, the influence of IL-6, matrigel, pre-treatment with an anti-mouse CD122-antibody (Ab) and/or the use of NOD/SCID-IL2-receptor-gamma-chain−/− (IL2−/−) mice, instead of conventional NOD/SCID mice, was evaluated. Tumor growth was monitored by a) multiparameter flow-cytometry (FACS; detection of human HLA-DR, HLA-A,B,C, CD45 and CD138), b) immunohistological detection of human CD138+ cells in tumor implants, BM and spleen and c) a fluorescence-based in vivo imaging system. L363 and RPMI8226 engrafted reliably at the injection site and in distant organs, independent of the experimental conditions (using IL-6, matrigel and/or pre-treatment with anti-CD122-Ab). L363 cells showed higher take- and metastases-rates compared to RPMI8226. The knock-down of NK cell activity, either by Ab-treatment or by genetic engineering, enhanced the tumor take rate with both MMCLs in 8/8 mice. With RPMI8226, the BM infiltration rate was 1.5–1.8% in all examined murine models. In contrast, BM-infiltration rates of L363 cells negatively correlated with the NK cell activity of the host: L363 cells metastasized particularly well to the BM when injected into anti-CD122-pretreated mice or intratibialy into IL-2−/− mice. Engraftment of circulating cells into the peripheral blood and spleen was found with both MMCLs irrespective of the mouse strain, pretreatment or implantation site. Matrigel or IL-6 showed no relevant engraftment effect, neither sc, nor with the intratibial approach. We conclude that the establishment of a metastasizing cell line-based human MM in vivo model was successfully pursued. We observed that the knock-down of NK cell activity was essential for both MMCLs, whereas the influence of matrigel or IL-6 treatment could be neglected. Furthermore, the intratibial approach optimized especially the L363 model in terms of infiltration rate to clinically relevant sites. With this optimized transplant approach, we aim to determine whether this allows MM engraftment using primary patient specimens, which should also enable us to test novel anti-myeloma agents.
Genetic Abnormalities Underlying B-Cell Development Are Characteristic for a New Subgroup of Childhood Precursor B-ALL with a Very Poor Prognosis
Authors: van Slegtenhorst Marjon , de Menezes Renee X , Pieters Rob , Boer M. L Den
Published: October 2019

Gene expression signatures are a promising tool to predict lineage and genetic subtypes of acute lymphoblastic leukemia (ALL) and other types of cancer. Using a 110 probe set signature, we correctly identified patients belonging to the 4 major subtypes of pediatric ALL (T-ALL, TELAML1-positive, hyperdiploid and E2A-rearranged ALL). In addition, we identified a novel subtype of precursor B-ALL. The signature of this group was most similar to true BCRABL-positive cases, whereas these cases did not have the BCRABL-translocation. We therefore describe this group as BCRABL-like. The existence of BCRABL-like patients was confirmed in an independent validation cohort of 107 patients. Most importantly, they represent 15–20% of precursor B-ALL cases with a highly unfavorable prognosis (5-year disease-free survival 60±11%) compared to other precursor B-ALL cases (84±4%; p=0.009), resembling that of true BCRABL-positive cases. To investigate the genetic abnormalities in BCRABL-like patients, we applied array-comparative genomic hybridization analysis. In 44 BCRABL-like patients, we found seven recurrent deletions. Most abnormalities were found in chromosome 9p, while other chromosomes affected included 20q, 22q11, 7p12, 11q, 13q14 and 19p13. Interestingly, many of these deleted regions harbor genes that play an important role in B-cell development. In total, 82% of BCRABL-like patients had abnormalities in one or more genes that are involved in B-cell development including PAX 5 (16 patients), VpreB1 (17 patients), Ikaros (17 patients), EBF1 (6 patients) and TCF3/E2A (3 patients). Bi-allelic loss was frequently found in VpreB1 and PAX5, including inactivating point mutations in exon 2 (V26G) in PAX5. The frequency of B-cell development gene deletions was also high in BCRABL-positive cases (80%), but was significant lower in a control group of precursor B-ALL, including TELAML1-positive, hyperdiploid, MLL-rearranged en B-other patients (36%, p=0.0002). In summary, these results indicate that the BCRABL-like group is a novel high-risk subtype of precursor B-ALL with a high percentage of deletions in B-cell development genes. Currently experiments are being done to determine the biological significance of the recurrent changes in the BCRABL-like group.
Transcription Factor Binding Sites Are Genetic Determinant of Retroviral Integration in the Human Genome
Authors: Cattoglio Claudia , Felice Barbara , Cittaro Davide , Miccio Annarita , Ferrari Giuliana , Recchia Alessandra , Luzi Lucilla , Mavilio Fulvio
Published: October 2019

Gamma-retroviruses and lentiviruses integrate non-randomly in mammalian genomes, with specific preferences for active chromatin, promoters and regulatory regions. Gamma-retroviral gene transfer vectors show a high frequency of insertional gene activation, and may induce neoplastic or pre-neoplastic clonal expansions in patients treated with genetically modified cells. An analysis of >10,000 integration sites of a Moloney leukemia virus (MLV)-derived vector in human CD34+ hematopoietic progenitors showed that genes involved the control of growth, differentiation and development of the hematopoietic and immune system are targeted at high frequency and enriched in integration hot spots, suggesting that the cell gene expression program is instrumental in directing MLV integration. To investigate the role of transcriptional regulatory networks in retroviral target site selection, we analyzed the distribution of transcription factor binding sites (TFBSs) flanking >4,000 MLV- and HIV-derived proviruses in human hematopoietic and non-hematopoietic cells. We show that MLV vectors integrate in genomic regions enriched in cell-type specific subsets of TFBSs, independently from their relative position with respect to genes and transcription start sites. Conversely, HIV vectors appear to avoid TFBS-rich genomic regions. Hierarchical clustering and a principal component analysis of TFBSs flanking retroviral integration sites in CD34+ and HeLa cells showed that TFBS subsets are vector- and cell type-specific. Analysis of sequences flanking the integration sites of vectors carrying mutations in their long terminal repeats (LTRs), and of HIV vectors packaged with a MLV integrase, indicates that the MLV integrase and LTR enhancer are the viral determinants of the selection of TFBS-rich regions in the genome. Chromatin immunoprecipitation analysis shows that transcription factors bind the LTR enhancers in the cell nucleus before integration, and may therefore synergize with the integrase in tethering retroviral pre-integration complexes to transcriptionally active regulatory regions. This study identifies TFBSs as differential genomic determinants of retroviral target site selection in the human genome, and indicates that gamma-retroviruses and lentiviruses have evolved dramatically different strategies to interact with the host cell chromatin. These differences predict a higher risk in using gamma-retroviral vs. lentiviral vectors for human gene therapy applications, independently from the design of the vector and the transgene expression cassette.
Escalating Dose Intravenous Methotrexate without Leucovorin Rescue during Interim Maintenance Is Superior to Oral Methotrexate for Children with Standard Risk Acute Lymphoblastic Leukemia (SR-ALL): Children’s Oncology Group Study 1991
Authors: Matloub Yousif , Bostrom Bruce C. , Hunger Stephen P. , Angiolillo Anne L , Cole Catherine , Thomson Blythe , Devidas Meenakshi , Heerema Nyla A. , La Mei K , Buckley Patrick J. , Carroll William L. , Winick Naomi , Sather Harland , Nachman James B , Gaynon Paul S.
Published: October 2019

A substantial number of relapses derive from the NCI standard risk population. CCG-1991 tested two components of the successful COG augmented “BFM” regimen (N Engl J Med1998; 338:1663) in this subset in a 2 × 2 factorial design. Patients received a 3-drug Induction: vincristine (V), pegylated asparaginase, dexamethasone (D), and intrathecal (IT) cytarabine and methotrexate (M). Patients with an unfavorable early marrow response by morphology (> 25% blasts at day 14, or >25% blasts at day 7 and >5% blasts at day 14) were assigned rescue daunorubicin and “augmented BFM” therapy. Other patients, termed rapid early responders (RER), received Consolidation (daily oral mercaptopurine (MP) and weekly IT M) and were randomized to receive either single or double delayed intensification (DI) phases, and D pulses, oral daily MP, weekly M and monthly V (Reg O) or V and intravenous (IV) escalating dose M with no leukovorin every 10 days for two months preceding and following the DI phase (Reg I).

Induction Consolidation Interim Maintenance Delayed Intensification (DI #1) Interim Maintenance Reg O Oral MP/M/D Reg O Oral 6MP/M/D Maintenance DI #2 Maintenance Reg I V/IV M Reg I V/IV M Maintenance DI #2 Maintenance

CCG-1991 enrolled a total of 3026 patients between 6/1/00 and 1/31/05 with 2078 eligible randomized non-T ALL patients. The overall 5-year event free (EFS) and overall survival (OS) for the randomized patients was 90.5% (SE = 1.0 %), and 96.0% (SE = 1.0%). We previously reported the double DI randomization (Blood2006; 108: abstract #146). Events were 71/1042 patients in Regimen I, and 100/1036 patients in Regimen O. For Reg I and Reg O, 5-year EFS was 92.1% and 88.7% (p = 0.02; RHR = 0.69 respectively. Comparison by type of events: isolated and combined marrow relapse (55 and 54), isolated CNS relapse (10 and 25), testes relapse (0 and 5), death in remission (1 and 5), secondary neoplasm (2 and 6), and other (2 and 4). The benefit was shown in both sexes. The overall survival was 96% on either arm. We therefore conclude that Reg I was superior for children with B-precursor SR-ALL with RER. We saw decreases in CNS and testes relapse with no decrease in marrow relapse. At this time, we find no difference in OS between the two arms.
Predictors of Osteopenia/Osteoporosis in Children with Acute Lymphoblastic Leukemia.
Authors: Athale Uma , Webber Colin , Barr Ronald
Published: October 2019

Background: Loss of bone mineral is a major problem in children with acute lymphoblastic leukemia (ALL), resulting in acute and chronic morbidity. About 30–40% of children with ALL will develop osteopenia/osteoporosis (OP) and about 10–15% will suffer from fractures. Identifying a population at high risk for OP is important to implementing a preventive strategy (e.g. biphosphonate therapy). So far predictors of OP in association with ALL in children are uncertain. Hence we undertook the following study to evaluate predictors of OP in children with ALL treated according to Dana-Farber Cancer Institute protocols.

Aim: To evaluate the relationship between lumbar spine bone mineral density (LS-BMD) Z scores in patients with ALL during maintenance therapy and the variables of age at diagnosis (< 10 vs. ≥ 10-years), risk group [Standard (SR) vs. high-risk (HR)], gender (male vs. female) and LS-BMD at diagnosis.

Methods: Children (≤ 18-years) diagnosed with ALL during the period 1995–2006 who were in first clinical remission, were included in the study. LS-BMD was measured using dual-energy X-ray absorptiometry (DEXA) at the time of diagnosis (n=88) and during the maintenance phase of therapy (n=119). The actual values of LS-BMD were expressed as age and gender matched Z-scores based on local population norms. Regression analyses were used to evaluate the risk of osteopenia, defined as LS-BMD Z score < -1.00, and osteoporosis, defined as LS-BMD Z score < -2.00. We evaluated the effect of age at diagnosis, gender, ALL risk category and LS-BMD at diagnosis on the LS-BMD during maintenance phase of therapy.

Results: Of the 119 patients, 19 (16%) were ≥ 10-years of age, 46 (39%) were girls and 41 (34.5%) had HR ALL. At diagnosis 29 of 88 (33%) patients had osteopenia and 6 (6.8%) had osteoporosis whereas, during maintenance therapy, 47/119 patients (39.5%) had osteopenia and 10 (8.4%) patients had osteoporosis. LS-BMD at diagnosis had a positive linear relationship with LS-BMD during maintenance therapy (Pearson correlation coefficient 0.721, p<0.001). Older children and children with HR ALL had a significantly higher risk of osteopenia compared to younger children (p=0.01) and children with SR ALL (p=0.019). Age and risk category were confounding variables since all children ≥ 10 years were classified as HR ALL. Gender by itself had no significant effect. However, the effect of age on the LS-BMD during maintenance phase was gender-dependent with older girls having lower LS-BMD compared to older boys.

Conclusions: Osteopenia and osteoporosis (OP) are common in children with ALL. Age over 10-years, female gender, HR ALL and lower LS-BMD at diagnosis are predictors of lower LS-BMD during the maintenance phase of therapy. Using these variables it is feasible to develop a predictive model to define the risk of OP during the maintenance phase of therapy. Larger prospective studies will better define this risk.
CD20 up-Regulation in Childhood B-Cell Precursor Acute Lymphoblastic Leukemia during Induction Treatment Translates into Higher Rituximab-Sensitivity.
Authors: Attarbaschi Andishe , Schumich Angela , Mann Georg , Haas Oskar A. , Gadner Helmut , Dworzak Michael
Published: October 2019

CD20 is expressed in about one half of childhood acute lymphoblastic leukemia (ALL) cases with B-cell precursor (BCP) origin. We recently observed that this phenotypic marker is further up-regulated in some patients during remission induction treatment containing corticosteroids for up to 5 weeks. To understand the impact of this phenomenon on the potential effectiveness of anti-CD20 immunotherapy (i.e., Rituximab), we studied 237 CD10-positive childhood BCP-ALL patients consecutively enrolled onto the trial AIEOP-BFM-ALL 2000. The analysis included the assessment of CD20 expression changes from diagnosis to the end of induction therapy and complement-induced cytotoxicity by CD20-targeting with Rituximab in-vitro. CD20-positivity significantly increased from diagnosis to the end of induction therapy with respect to the number of positive cases as well as to the levels of expression. After completion of induction therapy, one half of cases showed ≥ 90% CD20pos. leukemic cells, as opposed to 5% at diagnosis and 20% after 2 weeks of chemotherapy. Notably, up-regulation occurred in viable cells sustaining chemotherapy, most probably as a consequence of steroid-induced modulation of gene expression. Rituximab-cytotoxicity was significantly enhanced by CD20 up-regulation and depended on high expression levels. Importantly, CD20 up-regulation was frequent in high-risk patients (mainly poor prednisone responders), patients with high minimal residual disease levels at the end of induction therapy, and patients who suffered later from relapse, but not in TEL/AML1-positive cases. In conclusion, CD20 up-regulation is frequently induced in BCP-ALL during induction therapy and this translates into an acquired state of higher sensitivity to Rituximab.
Acute Erythroleukemia: An Analysis of 108 Patients Treated with Cytarabine-Containing Regimens at the M.D. Anderson Cancer Center.
Authors: Santos Fabio P.S. , Faderl Stefan , Garcia-Manero Guillermo , Koller Charles A. , Beran Miloslav , Borthakur Gautam , Bueso-Ramos Carlos , Keating Michael , Cortes Jorge , Kantarjian Hagop M. , Ravandi Farhad
Published: October 2019

Acute erythrocytic leukemia (AML-M6) is an uncommon subtype of acute myeloid leukemia (AML). It usually presents as a proliferation of dysplastic erythroid elements mixed with blasts of myeloid origin, although there are some rare cases of pure erythroid proliferation (DiGuglielmo’s disease). It has previously been associated with a poor prognosis. Here, we report the experience at the M.D. Anderson Cancer Center. From January, 1980 to May, 2006 there were 2992 patients with newly diagnosed AML (excluding APL), and 108 (4%) had the diagnosis of AML-M6. Diagnosis was based on morphological features according to the FAB classification, PAS-positivity by cytochemistry and immunophenotypical positivity for glycophorin A when available. Seventy (65%) were males; median age was 60 years (range 17 – 85). Antecedent hematological disorder (AHD) and/or MDS was present in 54 patients (50%) compared to 40% in other AML patients (p=0.056), and 27 (25%) patients had previously received treatment for other malignancies. The median white blood cell count at diagnosis was 2.5x109/L (0.3–45.1), hemoglobin 7.8 g/dL (2.5–12.6) and platelet count 40x109/L (3–22). Information about bone marrow dysplasia was available in 80 patients. Significant morphological dysplasia of erythroid lineage was found in 66 (83%), of myeloid lineage in 38 (48%) and of megakaryocytic lineage in 36 (45%) cases. Cytogenetic analysis was available in 106 (98%) cases, and was abnormal in 77 (71%) patients. The most common abnormalities were: Monosomy 5/Monosomy 7 in 50 (46.3%) and Trisomy 8 in 9 (8.3%). Poor risk cytogenetics were found in 46% of patients with AML-M6, comparing to 19% of the control group (other AML patients excluding APL) (p<0.001). All patients received induction chemotherapy containing cytarabine. Complete remission rates were 63% for AML-M6 patients, comparing to 58% for the control group (p = 0.285). Seventeen patients (15.7%) were refractory and 23 (21.3%) had induction death. Median disease free survival of AML-M6 patients was 31 weeks, compared to 49 weeks for the control group (p = 0.03). Median overall survival of AML-M6 patients was 33 weeks, compared to 42 weeks for the control group (p = 0.190). A total of 24 patients underwent allogeneic stem cell transplantation. Their median survival after transplantation was 32 weeks. We conclude that acute erythrocytic leukemia is a rare subtype of AML. It is commonly associated with a previous diagnosis of MDS and often presents with dysplastic features at diagnosis. It is frequently associated with poor-risk cytogenetics and a poor outcome.
NDRG1 Expression Is Inhibited in AML and Its Knockdown Attenuates Neutrophil Differentiation.
Authors: Tschan Mario P. , Shan Deborah , Laedrach Judith , Eyholzer Marianne , Leibundgut Elisabeth Oppliger , Baerlocher Gabriela , Tobler Andreas , Stroka Deborah , Fey Martin F.
Published: October 2019

The N-myc down-regulated gene 1 (NDRG1) is a stressed induced protein whose expression is associated with growth arrest and differentiation of tumor cells. Although the exact function of NDRG1 protein remains unknown, various studies support its role as a suppressor of tumor metastasis. In prostate, colon and breast cancer its expression is associated with a better disease prognosis and patient survival. In hematopoietic cells, NDRG1 was identified in a differential display screen for differentiation-related genes in human myelomonocytic U937 cells. In the present study, we sought to investigate the role of NDRG1 in myeloid differentiation. To this end we first evaluated NDRG1 mRNA expression in acute myeloid leukemia (AML; n=82) patient samples as well as in CD34+ progenitor cells (n=5) and neutrophils (n=6) of healthy donors using quantitative real-time RT-PCR. We found significantly higher NDRG1 mRNA levels in granulocytes as compared to CD34+ (p=0.0043) or AML blast cells (p<0.0001), whereas no significant difference between CD34+ progenitor and AML blast cells was seen (Figure A). Moreover, we found that NDRG1 mRNA levels were increased in 4/5 APL patients upon ATRA therapy. In contrast, the closest relative of NDRG1, NDRG2, did not show significantly different expression in these primary cells, thus indicating a unique role for NDRG1 in granulocyte differentiation. Next we examined NDRG1 expression using quantitative RT-PCR and Western blotting in two different cell line models for ATRA-induced neutrophil differentiation. ATRA treatment of NB4 and HT93 acute promyelocytic leukemia (APL) cells induced NDRG1 mRNA 2.3- and 14.3- fold, respectively. Increased NDRG1 mRNA expression was paralleled by an increase of NDRG1 protein as well as a decrease in c-myc protein. Earlier reports described that NDRG1 is also suppressed by c-myc suggesting that down-regulation of c-myc in our cell line models allowed for an increase of NDRG1. In line with these observations, lentivirus-driven short hairpin (sh)RNA-mediated silencing of NDRG1 diminished ATRA-induced neutrophil differentiation of NB4 and U937 cells as measured by CD11b, CD11c and CD18 surface expression. In NB4 NDRG1 knockdown versus non-targeting shRNA expressing cells mean fluorescent intensities (MFI) for CD11b, CD11c and CD18 upon six days of ATRA-treatment were 99±17 vs 146±7, 20±2 vs 32±10 and 19±2 vs 45±6, respectively (Figure B). Similarly, U937 NDRG1 knockdown versus control cells displayed the following MFIs for CD11b and CD18 upon neutrophil differentiation: 61±1 vs 102±2 and 11±4 vs 33±13, respectively. In conclusion, we report here for the first time an association of low NDRG1 levels with an immature hematopoietic cell phenotype. Using RNAi technology we further provide evidence that NDRG1 is functionally involved in neutrophil maturation.

Figure Figure
Survival after Complete Response to Treatment with Laromustine (CLORETAZINE®) in Elderly Patients with De Novo Poor Risk Acute Myelogenous Leukemia (AML).
Authors: O’Brien Susan , Schiller Gary , Stuart Robert Kenneth , Hudak Diana , Giles Francis
Published: October 2019

Background: Treatment options for patients with AML are limited. Poor outcomes are associated with increasing age, decreasing performance status, unfavorable cytogenetics, and the presence of comorbid conditions. Elderly patients ³ 65 years with AML have a short survival (median 2.4 months) (Lang 2005), and for older patients who receive treatment, 1-year survival is 5–25% (Menzin 2002, Kantarjian 2006, Burnett 2007). CR/CRp is a desired outcome from induction treatment and a requisite for meaningful survival. Laromustine (Cloretazine) is a novel sulfonylhydrazine alkylating agent which preferentially targets the O6 position of guanine resulting in DNA cross-links. Results of two sequential Phase II multi-center studies of single agent laromustine (CLI-033 and CLI-043) conducted in elderly untreated AML patients have been reported (Giles 2007, Schiller 2008). The purpose of this analysis is to assess survival for a combined group of patients from the two studies who achieved a response to laromustine (CR/CRp by IWG criteria [Cheson 2003]).

Methods: Eighty-five patients in study CLI-043 and 55 patients from a retrospectively identified de novo subset of CLI-033 who reasonably met the eligibility criteria for CLI- 043, received laromustine 600 mg/m2 as induction therapy (N=140). Poor risk was defined as the presence of at least one risk factor: age ³ 70 yrs, unfavorable cytogenetics, PS2, or pulmonary, cardiac, or hepatic comorbidity. 86% (120/140) pts had 2 or more risk factors. 16% (22/140) pts received a second induction of laromustine 600 mg/m2. 24% (34/140) patients received a consolidation cycle; per individual protocol, 15 pts received consolidation with laromustine 400 mg/m2 and 19 pts received cytarabine 400 mg/ m2. Kaplan-Meier analysis of survival was used to better understand the clinical benefit of CR/CRp.

Results: 52/140 (37%) pts achieved a CR/CRp; 44/140 (31%) pts survived ³ 30 days following CR/CRp. All but one patient achieved CR/CRp following first induction. Estimated Kaplan-Meier survival from time of response for these 44 patients is shown below. Six and 12 month survival for this subset is 58% and 44%, respectively. The 30-day mortality for the entire group is 14% (20/140).

Figure Figure

Conclusions: Early mortality, progressive disease, and short CR/CRp duration account for short median survival rates in leukemia studies. The 30-day mortality for this group compares favorably with published data of early death rates in older patients with multiple risk factors who receive induction chemotherapy (Appelbaum 2006, Kantarjian 2006). Relatively long survival in a subset of older patients with multiple risk factors who achieve a response to laromustine implies clinical benefit, and confirms the concept that obtaining a CR/CRp is in itself a benefit.
Late Relapses in Acute Myelogenous Leukemia: Analysis of Characteristics and Outcome.
Authors: Verma Dushyant , Faderl Stefan , O’Brien Susan , Pierce Sherry , Cortes Jorge , Vu Khanh , Freireich Emil , Keating Michael J. , Kantarjian Hagop M. , Ravandi Farhad
Published: October 2019

The majority of patients with acute myelogenous leukemia (AML) who achieve complete remission (CR) with initial chemotherapy will eventually relapse. Most relapses occur in the first three years and rare patients relapse after being in CR for more than 5 years. We have reviewed retrospectively 2347 patients with AML treated at M D Anderson Cancer Center from January 1980 to July 2008. Of the total cohort, 1366 patients achieved CR and among these 942 patients relapsed. We identified 11 patients (1.16%) who relapsed after being in CR >5 years; these patients are the focus of this analysis. There were 4 females and 7 males. Their median age was 66 years (range 37–79), and their median presentation white blood cell (WBC) count was 2.3 x109/L (range, 1.1–92.3). The FAB classification was M2 in 5 patients, M1, M4, M5, M6 in 1 patient each, and unknown in 2 patients. Initial cytogenetics were diploid in 6, del(7q) in one, miscellaneous in 1, and unavailable in 3 patients. Initial therapy was with combination of idarubicin (Ida) and cytarabine (Ara-C) in 4 patients (1 with additional fludarabine), amsacrine based in 3, daunorubicin (Dauno) single agent in 2 and other agents in 2 patients. All patients except one achieved CR after the first induction course; one patient needed 2 cycles of induction to achieve CR. None underwent an allogeneic stem cell transplant in first CR. The median duration of CR was 81 months (range, 60–137).

At the time of relapse, median WBC count was 4.4 x109/L (range 1.7–48.8). Karyotype at relapse was diploid in 2, del(5)del(7) in 1, del(6)del(7) in 1, trisomy 8 in 1, hyperdiploid in 2, add(2q) and add(6q) in 1 each and unavailable in 2 patients. The karyotype at relapse was different from the initial finding in 8 of 8 patients with available data at both time points. Treatment for relapse included Ida (or Dauno) with Ara-C in 8 patients (1 with additional fludarabine), and other agents in 3 patients. The response to treatment was CR in 4, partial remission in 2, resistant in 4 and unknown in 1. No patient underwent an allogeneic stem cell transplant in second CR. The median duration of the second CR was 2 months (range 0–37). The median survival after relapse was 6.4 months (range 1–39). Median survival from initial diagnosis of AML was 107 months (range 68–143). We conclude that late relapses in AML (>5 years after CR) are infrequent (1.16% of all relapses) and response to their subsequent therapy is poor; best responses occur with a regimen similar to the initial induction regimen. The karyotype at relapse is frequently different raising the question of a second AML versus relapse with the original clone.
Clinical and Serologic Interleukin 2 Receptor α Response to Interleukin-2 in CCG-2961, a Ranomized Phase 3 Trial for Pediatric Acute Myeloid Leukemia.
Authors: Lange Beverly , Yang Richard K , Gan Jacek , Hank Jaqueline A , Sievers Eric , Gerbing Robert , Alonzo Todd , Sondel Paul M
Published: October 2019

The Children’s Cancer Group conducted a randomized comparison of interleukin-2 (IL-2) to no further therapy (control) in pediatric patients with acute myeloid leukemia in first remission after 3 courses of dose intensive chemotherapy. To assess biologic activity of IL-2, serum samples for soluble IL-2 receptor alpha concentration (sIL-2rα) were collected before induction chemotherapy, and after completing chemotherapy but before (day 0), during (day 4) and after IL-2 (day 18) in those randomized to receive IL-2 and before chemotherapy and at the end of chemotherapy (day 0) and 28 days later in the controls. Of 385 eligible patients, 289 consented to participation in the randomization, and 172 submitted specimens. Patients randomized to IL-2 received IL-2 9 x 106 IU/m²/day by continuous infusion (CI) days 0–3 and 1.6 x 106 IU/m²/day CI days 8–17. At five years disease-free survival (DFS) is 51% + 9% in the IL-2 group and 58% + 8% in the controls (p =0.489), and respective overall survivals are 70% + 8% and 73% + 8% (p= 0.727). At diagnosis, median sIL-2rα concentration was 3342 pg/ml in the IL-2 group (N=34) and 3778 pg/ml in the controls (N=41) (p=0.947). In both groups there was a significant reduction in sIL-2rα concentration between the start and day 0 after chemotherapy. In the IL-2 group median sIL-2rα increased significantly from 1462 pg/ml on day zero to 13659 pg/ml on day 4 (p<0.001) and 8435 pg/ml on day 18 (p<0.001). In the control group day sIL-2rα was 1424 pg/ml on day zero and 1559 pg/ml on day 28. Despite evidence of substantial biologic activity as measured by significant increases in sIL-2rα concentration during IL-2 administration, IL-2 in this dose and schedule had no significant effects on DFS or survival. Furthermore, sIL2αr levels (the pretreatment sIL2rα level, the sIL2rα level obtained after chemotherapy or the level achieved after IL2 treatment) did not correlate with DFS or survival, or treatment-related toxicity.