bluuebunny/crossref_metadata_2025 huggingface dataset:
Difference in Outcome of Adolescents with Acute Lymphoblastic Leukemia (ALL) Enrolled in Pediatric (AIEOP) and Adult (GIMEMA) Protocols.
Authors: Testi Anna Maria , Valsecchi Maria Grazia , Conter Valentino , Vignetti Marco , Paoloni Francesca , Giona Fiorina , Moleti Maria Luisa , Nigro Luca Lo , Locatelli Franco , La Dogana Saverio , Specchia Giorgina , Fabiano Francesca , Recchia Anna , Mandelli Franco , Foa’ Robert , Masera Giuseppe
Published: October 2019

Progress in the treatment of acute lymphoblastic leukaemia (ALL) has led to better survival rates; however, children have had a greater benefit from improved treatment modalities than adolescent who show an overall lower event-free survival (EFS) compared to younger patients. Some differences in the clinical and biologic characteristics of adolescents compared to childhood ALL may partly account for the different outcome, but adolescents treated on pediatric ALL trials seem to have a significantly better EFS than those treated on adult trials. We retrospectively compared the results obtained in a series of 245 patients ranging in age from 14 to 18 years diagnosed and enrolled in specific Italian children and adult ALL trials, between 4/1996 and 10/2003. One hundred and fifty patients, from 30 pediatric centers, underwent the childhood AIEOP ALL 95 and 2000 protocols; the other 95, from 28 adult centers, were enrolled in the GIMEMA ALL 0496 and 2000 protocols. The AIEOP 95 and 2000 trials are BFM-like protocols with a 7 drug induction followed by risk-modulated post-remission therapy that includes high-dose MTX and reinduction for low and intermediate groups, and intensive blocks (high-dose MTX and cytarabine) for high-risk patients. Standard maintenance therapy is administered up to a total of 2 years. Cranial radiotherapy is limited to high-risk patients. Stem cell transplantation is planned for very high-risk patients. The GIMEMA regimens are instead based on an induction with high-dose anthracyclines (cumulative dose 550 mg/m2), high-dose cytarabine as consolidation and do not include high-dose MTX and the reinduction phase. Standard maintenance with vincristine + daunorubicin/cyclophosphamide pulses is given for 2 years. Cranial radiotherapy is administered to all patients. The main patients characteristics at diagnosis, in the two groups under examination, were comparable except for age: median age was 15 and 16 years, respectively in the AIEOP and GIMEMA trials.Poor risk cytogenetic translocations and T-immunophenotype were equally dinstributed. Adolescents in the AIEOP protocols had a higher CR rate (94% vs 89%) and a lower relapse rate (17% vs 45%) compared to the adolescents enrolled in the GIMEMA trials. The 2-year overall survival rate was 80% in the AIEOP protocols and 71% in the GIMEMA trials. Detailed results according to the different clinical and biologic features of the adolescents analyzed will be presented. The results of our comparative study indicate that adolescents enrolled in pediatric trials have a more favourable clinical outcome.
AMN107: In Vitro Profile of a New Inhibitor of the Tyrosine Kinase Activity of Bcr-Abl.
Authors: Mestan Juergen , Ellen Weisberg , Sandra Cowan-Jacob , Doriano Fabbro , Pascal Furet , Gabriele Fendrich , Gerard Goutte , Dominique Kempf , Markus Gaugler , James Griffin D. , Paul Manley W.
Published: October 2019

Imatinib mesylate (Gleevec®, Novartis Pharma AG), an inhibitor of the c-Abl/Bcr-Abl, c-Kit, and PDGFR kinases, is an effective therapy for CML, GIST and HES. However, advanced stage CML and Ph+ ALL patients frequently relapse due to the development of resistance. In up to 50% of cases, imatinib resistance is the result of the clonal expansion of cells expressing Bcr-Abl kinase domain mutants and more than 30 such mutations have been detected. Different mutations display varying degrees of resistance, and although some are sensitive to increased concentrations of imatinib, new Abl kinase inhibitors with higher potency against wild-type and imatinib-resistant mutants of Bcr-Abl could have substantial clinical utility. Rational drug design based on the structural information obtained from the crystal structure of the Abl-imatinib complex and medicinal chemistry optimization of drug-like properties, resulted in the discovery of AMN107. The potencies of AMN107 and imatinib were compared with cellular assays using CML lines (K562 and KU812F) and Ba/F3 cells expressing either ‘wt’- or imatinib-resistant mutants of Bcr-Abl. AMN107 effectively reduced the cellular Bcr-Abl autophosphorylation in K562, KU812F and Ba/F3 Bcr-Abl cells with IC50s of 42, 60 and 23 nM, respectively (Gleevec: 470, 399 and 231 nM) The proliferation of these cells was inhibited with IC50s of 11, 8 and 23 nM (Gleevec: 272, 80 and 634 nM). Autophosphorylation of the Bcr-Abl mutants M351T, F317L and E255V was reduced by AMN107 with IC50s of 33, 43 and 245 nM (Gleevec: 595, 818 and 6499 nM), resulting in inhibition of cell proliferation with IC50s of 30, 77 and 684 nM (Gleevec: 1285, 1583 and 6294 nM). No effect could be observed below 8 μM against the T315I and G250E bcr-abl mutants. Similar to imatinib, AMN107 inhibited the cellular tyrosine kinase activity of PDGFR-β (A31 cells) and c-Kit (GIST882 cells), with IC50s of 85 and 192 nM (Gleevec: 74 and 96 nM). Control (Ba/F3) cell proliferation was unaffected by 10 μM AMN107, and added IL-3 abrogated the antiproliferative effects on Bcr-Abl expressing Ba/F3 cells. Furthermore, AMN107 (< 10 μM), did not effect unrelated kinases, e.g. c-erbB2 (BT474), Ins-R (A14) and IGF-1R (NWT-21), in cellular assays, consistent with its specificity and lack of general cytotoxicity. Based on these in vitro assays and preclinical in vivo and pharmacokinetic data to be presented separately, AMN107 represents a promising new tyrosine kinase inhibitor for testing in human clinical trials.
Identification of Candidate Oncogenes in Acute Megakaryoblastic Leukemias with Gain of Chromosome 19.
Authors: Alvarez Sara , Largo Cristina , Blesa David , Gomez-Pineda Alfonso , Martin M. Carmen , MacGrogan Donal , Garcia Jose F. , Roncador Giovanna , Benitez Javier , Nimer Stephen D. , Cigudosa Juan C.
Published: October 2019

Acute Megakaryocytic leukemia is a rare subtype of AML that is often difficult to diagnose. Although GATA1 mutations are always found in AML-M7 associated with Down’s syndrome, such mutations are a rare event in adult AML-M7 leukemia. In 2001, we reported that Comparative Genomic Hybridization (CGH) identifies gain of chromosome 19 as part of the spectrum of nonrandom abnormalities characterizing megakaryoblastic proliferations, being its frequency underestimated when only conventional cytogenetics is used (1,2).

To identify candidate genes amplified and overexpressed on chromosome 19, we selected 10 cell lines derived from AML-M7 patients, that were previously characterized and classified by CGH according to their chr. 19 gain status. For each cell line, two hybridizations were performed: one with total RNA and one with DNA. Both hybridizations were done against the same c-DNA CNIO Oncochip, which allowed us to compare the results. As controls, 2 hybridizations with RNA obtained from CD34 cells selected from different leukapheresis were used to normalize the expression pattern from the cell lines.

First, we analyzed the expression values (normalized by the CD34+ cells) of the 201 c-DNA clones localized on chr. 19. The total number of overexpressed clones was higher in cell lines with gain or high level amplification of chr. 19 ( 38 to 79 of c-DNAs up) than in cell lines with normal chr. 19 or a discrete gain of 19q13.2 (29 to 31 of c-DNAs up). Second, examining the copy number changes and expression data allowed us to exclude some functionally interesting genes as MLL4 and to identify 36 clones that were recurrent and simultaneously gained and overexpressed. Eight out of these 36 clones showed at least three times greater expression in at least two cell lines. Six of these genes (KCNK6, APOC1, CD33, CEACAM5, CCNE1, and PPF1A3) are located on the q arm and two of them (NR2F6 and ASF1B) on the p arm. Interestingly, 80% of these 8 genes have at least one Gene Ontology annotated term relative to cell growth and/or maintenance. The dendogram of the unsupervised analysis of these 8 genes gives three arms that discriminate between 1) the cell lines with gain of chr. 19, 2) the cell lines with high level amplification of 19q, and 3) cell lines with normal chromosome19 or a discrete gain of 19q13.2. Moreover, the supervised analyses of the expression pattern of these 8 genes could identify significant differences among the four different states of chr.19 in the cell lines: high level amplification of 19q, gain of 19q13.2, gain of the whole 19, or normal 19, based on the expression of three genes: KCNK6, NR2F6, and APOC1. Further analyses, to identify the function of these genes in acute megakaryocytic leukemia cells are in progress.
Drug-Response Signature Predicts Outcome in Adult Acute Myeloid Leukemia and Associates Poor Response with Molecular Characteristics of Hematopoietic Stem Cells.
Authors: Heuser Michael , Wingen Luzie U. , Steinemann Doris , Cario Gunnar , von Neuhoff Nils , Tauscher Marcel , Bullinger Lars , Dohner Hartmut , Schlegelberger Brigitte , Ganser Arnold
Published: October 2019

Resistance to induction chemotherapy is of independent prognostic value in acute myeloid leukemia (AML). DNA microarrays were used to determine the gene-expression profile of AML blasts in 38 patients with good or poor response to induction chemotherapy. We selected an 11-sample training set, applying prediction analysis of microarrays (PAM) to devise a drug-response predictor which was tested on the remaining 27 samples and an independent set of samples recently published (Bullinger et al. 2004). Our drug-response predictor with 46 clones divided the 27 samples into two prognostic subgroups, the poor response group having a significantly shorter overall survival (P= .021). A subset of these 46 clones was sufficient to divide the published 62-sample test-set with intermediate risk cytogenetics into prognostically relevant subgroups (P= .028), adding prognostic information to that of known risk factors in multivariate analysis (hazard ratio, 2.8; 95 percent confidence interval, 1.4 to 5.8; P= .005). Thirteen of 39 drug resistance-enriched genes are known to be upregulated in hematopoietic stem/progenitor cells, and the expression pattern in normal CD34+ cells is highly correlated to the drug-resistance signature. This suggests that drug resistant AMLs show molecular features of hematopoietic stem/progenitor cells and can be identified by a characteristic gene-expression profile.
Pharmacology and Safety of SB-497115-GR, an Orally Active Small Molecular Weight TPO Receptor Agonist, in Chimpanzees, Rats and Dogs.
Authors: Sellers Teresa , Hart Timothy , Semanik Michael , Murthy Krishna
Published: October 2019

SB 497115-GR is a small molecular weight Tpo receptor (TpoR) agonist that has properties similar to thrombopoietin (TPO), primarily inducing proliferation and differentiation of megakaryocytes from bone marrow progenitor cells. SB-497115-GR is being developed for the treatment of thrombocytopenias, such as immune thrombocytopenic purpura. In vitro and in vivo studies have demonstrated that SB-497115-GR has very distinct species specificity. SB-497115 or other molecules in this class induced dose dependent STAT activation in platelets from humans and chimpanzees but not in platelets from laboratory animal species commonly used in drug safety studies. In order to demonstrate in vivo activity of SB-497115-GR, a single dose and 5 daily dose pharmacology and safety study in chimpanzees was conducted. To support initiation of clinical trials, a comprehensive package of toxicology studies was conducted including studies up to 14 days duration in rats and dogs. All procedures involving the care and use of animals in these studies were reviewed and approved by the appropriate Institutional Animal Care and Use Committees. Female chimpanzees (1–3/group) were administered vehicle or SB-497115-GR at doses of 0.1 to 10 mg/kg/day by oral gavage. For toxicology studies, SB-497115-GR was administered orally to rats (10/sex/group) by gavage at doses of 3 to 40 mg/kg/day and to dogs (3/sex/group) by capsule at doses of 3 to 30 mg/kg/day for 14 days. SB-497115-GR was well tolerated in chimpanzees, rats and dogs at all doses tested. In chimpanzees, no treatment related increases in platelet counts were observed after administration of single doses of up to 10 mg/kg or 5 daily doses of up to 3 mg/kg/day. However, following 5 daily doses of 10 mg/kg/day SB-497115-GR, there was a 1.3- to 2.4-fold increase in circulating platelet counts in 3 chimpanzees. A similar change in reticulated platelet counts was observed preceding this increase. In contrast, there was no effect of treatment for up to 14 days on platelet counts in rats or dogs. In conclusion, SB-497115-GR, an orally bioavailable small molecular weight agonist of the TpoR, has been shown to increase platelet counts in chimpanzees. These in vivo data confirm the in vitro data demonstrating the unique species-specific effects of this novel Tpo receptor agonist on platelets and were predictive of a pharmacodynamic effect currently being observed in human clinical trials.
Comparison of Three Tests for Confirmation of the Diagnosis of Heparin Induced Thrombocytopenia.
Authors: Messmore Harry L. , Choudhury Abdul M. , Fabbrini Nancy J. , Bird Mary L. , Moritz Thomas , Thottapurathu Lizzy , Wehrmacher William H. , Prechel Margaret , Walenga Jeanine M. , Crosby Thelma J. , Love Betty C. , Zarr Courtney A. , Griffin Beth E. , Balbale Sanober S.
Published: October 2019

In a recent two year period only 14% of platelet aggregation tests performed on 230 patients suspected of having heparin induced thrombocytopenia (HIT) were positive. The test (control platelet-rich plasma 0.14 mL, fresh heated patient serum 0.22 mL plus heparin 0.04 mL (1 Unit/mL final concentration) was performed using suitable controls on a commercial 4 channel aggregometer. 20% or greater increase in light transmission was a positive result. We then started a prospective study in which three different tests were done on the serum of each of 96 consecutive Hines Veterans Affairs Hospital patients who were clinically suspected of having HIT. The three tests were: platelet aggregation (PA) (same method as in the earlier study), C14 serotonin release (standard washed platelet method-SRA) and ELISA (GTI-PF4, Waukeshaw, Wisconsin), all performed on the same serum sample. The SRA and ELISA tests were performed on serum stored at −70°C. Blinded to the test results, a clinical score was determined from the record as HIGH (30% or greater fall in platelet count with recovery within 15 days off heparin), MEDIUM (same criteria but other cause present, or failure to recover platelet counts on continued heparin therapy) or LOW probability (failure to fall at least 30% or to recover from 30% or greater fall within 15 days observation). Results of the clinical scoring and laboratory tests were as follows: Clinical Probabilities: High (n=23), Medium (N=33), Low (n=40). Of the 56 who were High or Medium probability, 41% were High and 59% were Medium probability. Analysis of the results of the laboratory tests for sensitivity and specificity for each category revealed the following:

Clinical Probability PA SRA ELISA Sensitivity Specificity Sensitivity Specificity Sensitivity Specificity High and Medium (n=56) 19.6% 97.5% 10.7% 97.5% 10.7% 90% High (n=23) 30.4% 93.1% 17.4% 95.9% 8.7% 89%

Conclusions: Sensitivity of the PA test was greater than either of the other two tests, with specificities for the PA and SRA being nearly equal. The ELISA test was relatively insensitive and less specific than then other two.
Novel Lentiviral Vectors Displaying ‘Early-Acting-Cytokines’ Preferentially Promote the Survival and Transduction of NOD/SCID Repopulating Human Hematopoietic Stem Cells.
Authors: Verhoeyen E.L.S. , Wiznerowicz Maciej , Olivier Delphine , Izac Brigitte , Trono Didier , Dubart-Kupperschmitt Anne , Cosset Francois L.
Published: October 2019

A major limitation of current generation lentiviral vectors (LVs) is their inability to govern efficient gene transfer into quiescent target cells which hampers their application for hematopoietic stem cell gene therapy. Human CD34+ cells that reside into G0 phase of the cell cycle and thus are quiescent, are indeed higly enriched in hematopoietic stem cells. Here, we designed novel lentiviral vectors that overcome this type of restriction by displaying early-acting-cytokines on their surface. Presentation of a single cytokine, thrombopoietin (TPO), or co-presentation of TPO and stem cell factor (SCF) on the lentiviral vector surface improved gene transfer into quiescent CD34+ cord blood cells by 45-fold and 77-fold, respectively, as compared to conventional lentiviral vectors. Moreover, these new LVs preferentially transduced and promoted the survival of immature resting cells rather than cycling CD34+ cells. Most importantly, the new early-cytokine-displaying lentiviral vectors allowed highly efficient gene transfer in CD34+ immature cells with long-term in vivo NOD/SCID mice repopulating capacity, a hallmark of bona fide HSCs. In conclusion, the novel ‘early-acting cytokines’ displaying LVs described here provide simplified, reproducible gene transfer protocols that ensure efficient gene transfer in hematopoietic stem cells. As such, these novel reagents bring us one step closer to selective in vivo gene therapy.
Directing Oncogenic Fusion Genes into Stem Cells Via an Scl Enhancer.
Authors: Eguchi Mariko , Eguchi-Ishimae Minenori , Green Anthony R. , Enver Tariq , Greaves Mel
Published: October 2019

Chimeric fusion genes generated by chromosomal translocation are a consistent feature of leukemias and some other cancer types, including sarcomas, and usually specific to particular subtypes of leukemia or tumors. One favoured explanation for this selective transformation is that it may reflect context-dependent oncogenic functions in particular stem cells and their progeny rather than highly restricted origins of fusion gene recombination itself. We have assessed this proposition by directing the expression of fusion oncogenes in stem cells in vivo.

TEL-TRKC is a fusion gene generated by chromosomal translocation and encodes an activated tyrosine kinase. Uniquely, it associates with both rare solid tumors (congenital fibrosarcoma, congenital mesoblastic nephroma and secretory breast carcinoma) and acute leukemia, but a single exon difference (in TEL) between the two phenotypes. We expressed the two TEL-TRKC variants in mice using the 3′ regulatory element of SCL that is selectively active in a subset of mesodermal cell lineages including endothelial and hematopoietic stem cells and progenitors. The leukemia form of TEL-TRKC (- exon 5 of TEL) enhanced hematopoietic stem cell renewal and initiated leukemia in transgenic mice. In contrast, the TEL-TRKC solid tumor variant (+ TEL exon 5) elicited an embryonic lethal phenotype around day 12.5 (E12.5) with impairment of both angiogenesis and hematopoiesis indicative of an effect at the level of the hemangioblasts. These data indicate that oncogenic fusion proteins similarly expressed in a hierarchy of early stem cells can have selective, cell type specific developmental impacts dependent upon their intrinsic molecular properties.
The Roles of Stat3 and ERK in G-CSF Signaling.
Authors: Kamezaki Kenjirou , Shimoda Kazuya , Numata Akihiko , Yoshie Masumi , Yamamoto Masahiro , Takeda Kiyoshi , Matsuda Tadashi , Akira Shizuo , Ogawa Katsuhiro , Harada Mine
Published: October 2019

G-CSF specifically stimulates the proliferation and differentiation of cells that are committed to the neutrophil-granulocyte lineage. We examined the role of stat3 in G-CSF signaling using the Cre-loxP recombination system. Although Stat3 was thought to be essential for the transduction of G-CSF-induced cell proliferation and differentiation signals, mice deficient for Stat3 in hematopoietic cells show neutrocytosis and infiltration of cells into the digestive tract. The number of progenitor cells in the neutrophil lineage is not changed, and G-CSF-induced proliferation of progenitor cells and prolonged neutrophil survival were observed in Stat3-deficient mice. In hematopoietic cells from Stat3-deficient mice, trace levels of SOCS3, a negative regulator of granulopoiesis, were observed, and SOCS3 expression was not induced by G-CSF stimulation. Stat3 null bone marrow cells displayed a significant activation of ERK1/2 under basal conditions, and the activation of ERK was enhanced and sustained by G-CSF stimulation. Furthermore, the augmented proliferation of Stat3-deficient bone marrow cells in response to G-CSF was dramatically decreased by addition of a MEK1 inhibitor. These results indicate that Stat3 functions as a negative regulator of G-CSF signaling by inducing SOCS3 expression, and ERK activation is the major factor responsible for inducing the proliferation of hematopoietic cells in response to G-CSF.
Prevalence of Microbially Contaminated Hematopoietic Stem Cell Products.
Authors: Champlin Richard E. , Loberiza Fausto R. , Eapen Mary , Rizzo J. Douglas , Bredeson Christopher N. , Wagner John E. , Horowitz Mary M. ,   on behalf of the Histocompatibility Alternative Stem Cell Sources Working Committee of the Center for International Blood Marrow Transplant Research (CIBMTR)
Published: October 2019

In 2001, the Docket Report from the Food and Drug Administration expressed concerns regarding the potential of microbially contaminated hematopoietic stem cell products to produce morbidity and mortality in transplant recipients. This concern was the basis for development of regulatory standards for hematopoietic stem cell products.

We surveyed a total of 2972 patients at 121 U.S. transplant centers that registered patients with the CIBMTR in the years 2000 and 2001. Information regarding microbial contamination of infused grafts was obtained from 94 transplant centers (80% response rate) for 2312 patients. 52 (2%) of 2286 infused grafts tested were culture positive for bacterial or fungal organisms. The microbial isolates included: coagulase negative staphylococcus (56%), gram negative organisms (15%), coagulase positive staphylococcus (10%), gram positive rods (10%), streptococcus (8%), and fungus (1%). Prophylactic antibiotics targeted at the contaminant were given to 17 of the 52 recipients of contaminated grafts. Antibiotic regimens included vancomycin alone (76%), aminoglycosides and vancomycin (12%), or cephalosporin and vancomycin (12%). 47 (50%) of the centers that participated have existing policies regarding contaminated products. Patients with non-malignant disorders or who received bone marrow were more likely to have a contaminated graft. No differences in age distribution, sex, race, type of transplant (allogeneic vs autologous) and year of transplant were noted between recipients of contaminated and non-contaminated grafts. The unadjusted 100-day survival of persons receiving contaminated grafts was 86% (95% Confidence Interval [CI] 72–93%) versus 81% (95% CI 80–83%) among those receiving non-contaminated grafts, p=0.35.

In summary, about 2% of hematopoietic stem cell products infused for allogeneic or autologous transplantations in U.S. centers will test positive for microbial contamination, but such contamination does not increase posttransplant mortality. The absence of significant 100-day mortality among patients infused with contaminated grafts suggests that stringent regulatory policies regarding the use of contaminated hematopoietic cell products may not be indicated.
Four Years of Follow-Up of 1027 Patients with Late Chronic Phase (L-CP), Accelerated Phase (AP), or Blast Crisis (BC) Chronic Myeloid Leukemia (CML) Treated with Imatinib in Three Large Phase II Trials.
Authors: Silver Richard T. , Talpaz Moshe , Sawyers Charles L. , Druker Brian J. , Hochhaus Andreas , Schiffer Charles A. , Guilhot Francois , Goldman John M. , Smith B. Douglas , Mone Manisha , Kranhnke Tillmann , Kantarjian Hagop M.
Published: October 2019

Background

This report updates the results of 3 large phase II studies of the orally available BCR-ABL tyrosine kinase inhibitor imatinib for patients (pts) in AP, BC and late chronic phase (L-CP) CML failing prior interferon therapy (Kantarjan et al, ASH 2003; Talpaz et al, ASH 2003).

Methods

Between August 1999 and June 2000, 1027 pts were enrolled in phase II trials for CML in L-CP (n=532), AP (n=235) or BC (n=260). Pts in L-CP were treated with 400 mg/day and pts in AP or BC with either 400 or 600 mg/day. Dose escalation up to 800 mg/d was allowed in the late-chronic phase study. Pts with a confirmed diagnosis of AP (n=181), BC (n=229) and late-chronic phase (n=454) were evaluated for efficacy. All pts were evaluated for safety. The median time from initial diagnosis to study entry was 32 months for L-CP pts.

Results

As of 31-Jul-03, 5% patients with BC, 25% of CML-AP and 64% of L-CP patients still remain on treatment.

At the recommended dose of 600 mg, an estimated 40% (AP) and 7% (BC) of patients remained progression-free at 36 months, and an estimated 55% (AP) and 14% (BC) patients were alive at 36 months after initiation of imatinib. The 3-year survival rates for pts with AP with a major cytogenetic response at 3 months were 85% vs. 52% for pts with no response (p<0.001). In L-CP patients with a median follow-up of 40 months, 65% of patients achieved a major cytogenetic response, which was complete in 52%. The cytogenetic responses were durable with an estimated 82% of the pts in continuos major cytogenetic response at 3 years. The estimated rates of progression-free survival and overall survival at 3 years were 80% and 88%. Pts with at least a minor cytogenetic response at 6 months ≤65% Ph+ cells) had an estimated 3-year survival rate of 96% vs. 86% for pts with a ( minimal response and 81% for pts with no cytogenetic response (p<0.001).

Conclusion

In large phase II studies, continuous imatinib treatment is safe and has improved progression-free survival of patients at all stages of CML. Responses to imatinib are durable and are predictive of long-term outcomes. These results will be further updated at the meeting using a data base lock planned for 20-Sept-04 (using data collected up to 31-July-04, i.e. more than 4 years after the last pts enrollment).
Avascular Necrosis of Bone after Therapy for Multiple Myeloma: A Study of 561 Consecutive Patients.
Authors: Anaissie Elias J. , Talamo Giampaolo , Angtuaco Edgardo , Walker Ronald C. , Dong Li , Zangari Maurizio , Tricot Guido , Barlogie Bart
Published: October 2019

Purpose: To assess the prevalence, time of onset, risk factors, and outcome of avascular necrosis (AVN) of bone in patients with multiple myeloma undergoing antineoplastic therapy.

Patients And Methods: We reviewed the medical records of 561 myeloma patients (553 evaluable) enrolled in a treatment protocol consisting of dexamethasone-containing induction chemotherapy, autologous stem cell transplantation (ASCT), consolidation chemotherapy and maintenance with alfa-interferon. Patients were randomized at enrollment to receive thalidomide (269 pts) or no thalidomide (284 pts) throughout the study period.

Results: At a median follow-up of 33 months (range, 5–114 months), AVN of the femoral head (s) developed in 49 of 553 patients (9%). Median time to onset of AVN was 12 months (range, 2–41 months). Three risk factors for AVN were identified by multivariate analysis: cumulative dexamethasone dose (p=0.0006; O.R. 1.028; 95% C.I. 1.012–1.044 per 40 mg dexamethasone), male gender (p=0.009; O.R. 0.390; 95% C.I. 0.192–0.790) and younger age (p=0.0122; 95% O.R. 0.961, C.I. 0.934–0991/year). AVN-related pain and limited range of motion of the affected joint were present in only 9 and 4 patients, respectively. Fluorine-18 fluorodeoxyglucose positron emission tomography (FDG-PET) failed to detect abnormal uptake in the AVN-affected bones. Thalidomide-treated patients had a prevalence of AVN similar to that of the control group (8 % vs.10%, respectively; p=0.58).

Conclusion: AVN is a rare and usually asymptomatic complication in myeloma patients undergoing antineoplastic treatment with dexamethasone-containing regimens and ASCT. Cumulative dexamethasone dose, male gender and younger age, but not thalidomide, increase the risk of AVN. FDG-PET does not contribute to the diagnosis of this complication
Serum Transferrin Receptor Value as a Universal Indicator of Erythropoietic Activity.
Authors: Lee Young Soo , Kim Chul Soo , Choi Jong Weon
Published: October 2019

The serum transferrin receptor (sTfR) is thought a sensitive and quantitative parameter of tissue iron deficiency as well as an indicator of erythropoietic activity. This study was aimed at the verification of a hypothesis that sTfR is a general indicator of erythropoiesis regardless whatever the cause is. A total of 173 patients in heterogeneous diseases who underwent bone marrow study as a workup for anemia were measured for sTfR, reticulocyte maturity index (RMI), erythroid element proportion of bone marrow cells, and other hematologic parameters (hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin concentration, red cell distribution width, absolute reticulocyte count). By immunoenzymometric method sTfR was measured using IDeATMcTfR kids (Orion Diagnostica, Orion, Finland). Reticulocyte count and proportion was measured manually by one expert examiner after standard blood smear and stain. Reticulocyte subpopulation was automatically analyzed by flow cytometry using R-3000 TM (Sysmex, TOA, Japan). RMI was calculated from the equation of (medium fluorescent reticulocyte fraction + high fluorescent reticulocyte fraction) X 100 / low fluorescent reticulocyte fraction. Correlation analysis was done among the variables including sTfR, RMI, erythroid element proportion of bone marrow cells, and other hematologic parameters using SAS 6.12 soft ware. The analysis was carried out for the whole 173 patients to see the general trends and repeated for 4 groups of disease category, arbitrarily divided to group 1 (n=33, iron deficiency or or disease with no predisposition to anemia of chronic disease), group 2 (n=53, hematologic malignancies), group 3 (n=44, solid tumors), and group 4 (n=43, chronic or infectious disease) to see if the trends may be affected by specific diseases. The results showed a solid correlation of sTfR with RMI as well as erythroid precursors in bone marrow, not only in the whole patient population (e.g. sTfR vs RMI, R=0.587, p=0.0001) but also in individual groups (e.g. sTfR vs RMI, R=0.48, p=0.005 in group 1, R=0.69, p=0.0001 in group 2, R=0.58, p=0.0001 in group 3, R=0.81, p=0.0001 in group 4). These findings indicated the significance of sTfR is valid under any clinical setting as a universal indicator of hematopoietic activity. The sTfR can be used as a useful parameter for monitoring of erythropoiesis in a variety disease.
An Extra Ordinary Respone of Castleman’s Disease to Rituximab.
Authors: Abdou Sherif , Salib Hayman
Published: October 2019

Castleman’s disease is a very rare non-neoplastic lymphoproliferative disorder of unknown cause that is diagnosed on a pathological basis. It is often associated with autoimmune diseases, and diagnosed in the context of human immunodeficiency virus (HIV) positive patients. No standard treatment has been developed for this rare disease.

CASE REPORT: This 54-year-old male presented with nausea and vomiting. He has a past medical history of hypertension and chronic glomerulonephritis. His laboratory results showed acute renal failure. Hemodialysis was started prior to further work up. A chest x-ray showed bilateral hilar lymphadenopathy that was later confirmed by computerized tomography (CT) of the chest. A mediastinoscopy and mediastinotomy were performed. Mediastinal lymph node biopsy showed findings consistent with Castleman’s disease including atrophic follicles with hyalinized vessels. Remission was obtained for one year by prednisone and cyclophosphamide, but the patient relapsed with multiple lymphadenopathy. Biopsy confirmed the recurrence of Castleman’s disease. Rituximab was added to cyclophosphamide as a treatment regimen and for more than two years the patient has done extremely well. His kidney function returned to his baseline, hemodialysis was stopped, and the patient has long-term remission on CT scan and postron emission tomography (PET).

DISCUSSION: Castleman’s disease is lymphadenopathy of unknown cause. About 400 patients have been reported to date. It is diagnosed microscopically by lymphadenopathy with angiofollicular hyperplasia and plasma cell infiltration. It has a rapid progressive and often fatal course. Traditional treatment is chemotherapy. A new trend in the treatment of Castleman’s disease is Rituximab which is a monoclonal antibody that showed success in HIV positive patients with Castleman’s disease. We believe that this is the first reported case of long-term remission of Castleman’s disease in an HIV-negative patient treated with Rituximab and chemotherapy.
Injury of Endothelium during the Cytostatic Therapy of AML Patients.
Authors: Chervontseva Alevtina , Savchenko Valeri , Romanov Yuri
Published: October 2019

Background: At present, program chemotherapy represents the main approach to the treatment of patients with acute myeloid leukemia (AML). Chemotherapy is directed to the eradication of tumor clones and restoration of normal hematopoiesis. Using “7 + 3” protocol, including standard doses of cytosine arabinoside (ara-C) and daunorubicin (DNR) it is possible to obtain complete remission in 60–70% of patients. However, several vascular complications may take place during chemotherapy that could be related to vascular endothelium injury.

Aims: To investigate the effects of ara-C and DNR on cultured human vascular endothelial cells (ECs) and to confirm endothelial injury by the detection of circulating ECs.

Methods: Experiments were performed on cultured human umbilical vein ECs; cytostatic drugs were studied at therapeutical concentrations ranging from 1 ng/ml to 1 mg/ml. Effects were screened using phase-contrast microscopy, cell viability test, BRDU incorporation, immunocytochemistry, and flow cytometry.

Results: At various concentrations, ara-C and DNR induced EC monolayer changes associated with either cytostatic or cytotoxic action. DNR provoked irreversible cell changes and monolayer damage; the action of ara-C leaded to elimination of mitotic ECs from self-renewing monolayer and to significant decrease of its density. Simultaneously, remaining ECs increased the expression of the cell adhesion molecules (P- and E-selectins, VCAM-1, ICAM-1, and PECAM-1) and enhanced their adhesiveness to blood mononuclear cells. Comparing the level of ECs circulating in peripheral blood of healthy donors and AML patients undergoing chemotherapy, we have find remarkable differences. In the studied group, the concentration of circulating ECs could be as high as 250–750 cells per 1 ml of blood.

Conclusions: Obtained results allow to conclude that cytostatic therapy may cause systemic injury of the vascular endothelium related to functional cell alterations and/or massive EC loss.
Characterization of the Interaction between Cleaved High Molecular Weight Kininogen (HKa) and Tropomyosin (TM) on Endothelial Cells.
Authors: Qi Xiaoping , McCrae Keith R.
Published: October 2019

We previously reported that two chain human high molecular weight kininogen (HKa) inhibited angiogenesis by selectively inducing apoptosis of proliferating endothelial cells (Zhang et al. FASEB J., 2000). This activity appears to depend upon binding of HKa to endothelial cell tropomyosin, as it was completely inhibited by anti-tropomyosin monoclonal antibody (mAb) TM-311. mAb TM-311 also blocked the high-affinity Zn2+-dependent binding of HKa to both purified tropomyosin (TM) and proliferating endothelial cells (Zhang et al., PNAS 2003). However, endothelial cells express several different isoforms of TM, and the isoform(s) expressed on the endothelial cell surface, and the regions within it that bind HKa are unknown. To identify which isoform of TM is expressed on the surface of endothelial cells, human umbilical vein endothelial cells (HUVEC) were briefly exposed to a buffer containing 0.05 M glycine, 0.1 M NaCl, pH 3.0, and the resultant eluate was probed with isoform specific anti-tropomyosin antibodies that recognize TMs 1–5. Only TM5 was detected, with approximately 2.5-fold more TM5 eluted from proliferating versus confluent cells. Northern blot analysis also showed that the expression of TM5 mRNA was higher in the proliferating versus confluent cells. To locate the binding site for HKa in human TM, hTM3 was digested by CNBr, producing three fragments containing amino acids 10–127, 141–200 and 208–270. These were incubated with HKa and the mixture then passed through a Superose 6 gel filtration column. Analysis of the first peak that eluted from the column by tricine gel electrophoresis revealed two bands, the smaller of which contained the N-terminal CNBr-derived TM fragment (aa 10–127), and the larger of which contained HKa. Taken together with our previous observation that HKa binds with similar affinity to TM isoforms 1–5, suggesting that it binds to a homologous region among these proteins, these findings suggest that the binding site for HKa resides within the homologous region within the N-terminal TM fragment, and is likely contained within amino acids 81 to127aa of TM3 (Figure). Since these results suggest that at least hTM5 is non-covalently associated with the endothelial cell surface, we have begun to explore the nature of this interaction. Endothelial cell surface proteins were cross linked using the membrane-impermeable cross-linker, BS3, and cell extracts were then immunoblotted using the anti-hTM5 antibody LC-1. These result revealed native hTM5 (~30 kD), as well as a new band of ~60 kD, suggesting an association of hTM5 with a cell surface protein of approximately equal size. In conclusion, our results suggest that the anti-endothelial cell activity of HKa is mediated through binding to cell surface hTM5, via a homologous region of this protein shared with other non-muscle tropomyosins.
ALFA-9000 Trial - An Analysis of Cytogenetic Subsets: Significant Long-Term Benefit of Early Induction Reinforcement Is Observed in CBF-AML Only.
Authors: Gardin Claude , Chevret Sylvie , Charrin Christiane , Thomas Xavier , Bastard Christian , Lai Jean-Luc , Maarek Odile , de Botton Stephane , Bordessoule Dominique , Tilly Herve , de Revel Thierry , Terre Christine , Preudhomme Claude , Bauters Francis , Michallet Mauricette , Fiere Denis , Castaigne Sylvie , Fenaux Pierre , Dombret Herve
Published: October 2019

We have recently reported the results of the ALFA-9000 randomized trial in which younger adults with AML had a significant better long-term relapse-free interval when receiving a timed-sequentially reinforced induction (arm C) as compared to either standard 3+7 (arm A) or double (arm B) induction (Castaigne et al, Blood 2004). Both arms B and C included the same induction reinforcement with mitoxantrone and intermediate-dose cytarabine (ID-AraC) systematically administered at day 20 in arm B or at day 8 in arm C. Although patients aged between 15 and 65 years were included in this study, the gain in relapse incidence associated with arm C was observed in those less than 50 years of age only. We report here the outcomes of cytogenetic subsets, according to the arm C versus arm A/B stratification. Overall, 405 karyotypes were available (68%) and classified according to the SWOG classification as 56 favorable (CBF-AML, 46/56 under the age of 50 years), 215 intermediate (including 194 normal), 91 unfavorable, and 43 of unknown significance. Outcome of unfavorable and unknown subsets did not differ significantly in C versus A/B. Patients with complex karyotype or chromosome 7, 5q, or 3q abnormalities, as well as those with 11q23 abnormalities, had a similar outcome whatever their induction arm. Similarly, no benefit of early reinforcement was observed in the intermediate group, nor in patients with a normal karyotype even after adjustment on WBC or when considering only those aged less than 50 years. In the favorable subset, patients with either t(8;21) (N=30) or inv(16) (N=26) had identical long-term outcome. Contrary to other subsets, this outcome differed strikingly among induction arms with better disease-free survival in arm C (64% versus 28% at 5 years, P=.05 after adjustment on WBC) with a similar trend observed for event-free survival. The value of higher than standard doses of cytarabine in CBF-AML patients, either during induction or as post-remission therapy, has been demonstrated by several cooperative groups. Our results do suggest that early introduction of ID-AraC is also associated with long-term benefit in this favorable subgroup of patients, even greater when made at day 8 instead of day 20. We will evaluate prospectively this induction approach in the next French Intergroup CBF-AML study, in which all patients will then receive high-dose cytarabine during post-remission therapy.
Effects of Single-Dose BAY 59-7939 - An Oral, Direct Factor Xa Inhibitor - in Subjects with Extreme Body Weight.
Authors: Kubitza Dagmar , Becka Michael , Zuehlsdorf Michael , Mueck Wolfgang
Published: October 2019

Currently, agents used for the prevention of venous thromboembolism, such as low molecular weight heparins and pentasaccharides, require dose adjustments for patients in extreme weight categories. BAY 59-7939 is a novel, oral, direct Factor Xa (FXa) inhibitor currently undergoing clinical development for the prevention and treatment of thromboembolic disorders. In this single-center, randomized, single-blind, placebo-controlled, parallel-group study, the influence of extremely low and high body weight on the safety, tolerability, pharmacodynamics and pharmacokinetics of a single 10 mg dose of BAY 59-7939 administered with food was investigated. In total, 48 healthy male and female subjects aged 20–54 years and in three different weight groups (≤ 50, 70–80, and >120 kg) were evaluated; 12 received placebo and 36 received BAY 59-7939. The 70–80 kg and >120 kg groups were gender-balanced (six men and six women in each group); the ≤ 50 kg group comprised of 12 women. Overall, BAY 59-7939 was well tolerated. No serious adverse events were reported; all events were of mild to moderate intensity and resolved 7 days after drug administration. The occurrence of adverse events was similar among the weight groups. No adverse events were reported in the placebo group. FXa was inhibited by BAY 59-7939 in all three weight groups, with a maximum inhibition (Emax) of 46.8%, 45.8%, and 41.7% in the ≤ 50, 70–80, and >120 kg groups, respectively; maximum inhibition was observed about 2–3 hours after BAY 59-7939 administration in all three weight groups. AUC for inhibition of Factor Xa activity was comparable for all volunteer groups regardless of the body weight. BAY 59-7939 prolonged prothrombin time (PT), which was less pronounced in the higher weight groups: 1.7 times baseline (tb), 1.6 tb, and 1.5 tb in subjects ≤ 50, 70–80, and >120 kg, respectively. The pharmacodynamic parameters have returned to baseline within 24 hours in all weight groups. Placebo treatment had no effect on PT. The clotting tests activated partial thromboplastin time and HepTest were also distinctly prolonged after drug administration in all weight groups; the prolongation was slightly more pronounced in subjects in the ≤ 50 kg group than in the other weight groups. The bioavailability of BAY 59-7939 as a 10 mg single dose, in terms of AUC of plasma concentrations, was similar in all three weight groups (P=0.205). However, peak plasma concentrations (Cmax) were 24% higher in subjects with ≤ 50 kg body weight compared with normal-weight subjects. For subjects >120 kg, Cmax was similar to normal-weight subjects. No pharmacokinetic differences between men and women were detected in the normal-weight and >120 kg groups. In conclusion, these data demonstrate that only small influences of body weight on pharmacodynamic or pharmacokinetic parameters are observed which require no dose adjustment of BAY 59-7936 in subjects with extreme body weight.
Anticoagulant and Anti-Inflammatory Effects of Recombinant Human Antithrombin in Man after LPS Challenge.
Authors: Jilma Bernd , Leitner Judith M. , Mayr Florian B. , Firbas Christa , Reiter Rosemarie A. , Kalhs Peter
Published: October 2019

Background: Antithrombin (AT) had beneficial effects on mortality of septic patients in an a priori defined subpopulation of the Kypersept trial, which received no concomitant administration of heparin.

Objectives: We hypothesized that recombinant human (rh)AT (without concomitant heparin) has anticoagulant properties and may decrease cytokine production in a well standardized model of human endotoxemia.

Methods: This study was randomized, double-blind, placebo-controlled in parallel groups in 30 healthy male volunteers. The active treatment groups received bolus primed continuous infusion of rhAT (recombinant human Antithrombin) to increase AT-levels to 200% and 500% iv. before infusion of 2ng/kg endotoxin

Results: Infusion of rhAT rapidly decreased neutrophil (p<0.01) and monocyte counts (p<0.05) before LPS-challenge, demonstrating that rhAT directly interacts with these leukocyte subsets. rhAT exhibited intrinsic anticoagulant efficacy ex vivo as shown by thrombelastography ((p<0.01). More importantly, in vivo thrombin formation decreased in a dose dependent fashion as measured by prothrombin fragment (F1+2) and thrombin antithrombin complexes (TAT). rhAT significantly decreased interleukin-6 (IL-6) release by 40%.

Conclusion: rhAT exhibited intrinsic anticoagulant effects, which are stronger than the minor effects of drotrecogin alfa (rhAPC; Blood2003;102:2093ff) in this model of tissue factor triggered coagulation. Effects of rhAT on leukocytes and inhibition of IL-6 release were not previously observed with various different synthetic or natural anticoagulants in this model and therefore represent specific pharmacodynamic properties of rhAT. The demonstrated anticoagulant and anti-inflammatory properties of rhAT may have beneficial effects in critically ill patients, and may explain the efficacy of rhAT in septic patients without concomitant heparin administration.