bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prognostic Role of Glutathione S-Transferase (GST) Polymorphisms in Acute Myeloid Leukemia.
Authors: Voso Maria Teresa , Hohaus Stefan , Guidi Francesco , Fabiani Emiliano , D’Alo’ Francesco , Groener Silia , Spaeth Daniela , Dohner Konstanze , Leone Giuseppe , Dohner Hartmut , Schlenk Richard F. 
Published: April 2020

Abstract
Glutathione S-transferases (GST) are phase II detoxification enzymes, involved in the metabolism of carcinogens and anti-cancer drugs. Since GSTs have been also shown to interact with kinase complexes during oxidative or chemical stress-induced apoptosis, their polymorphic variants may account for differences in outcome following chemotherapy between individuals. Studying 106 patients (pts) with acute myeloid leukaemia (AML), we have shown that individuals with a GSTM1 and/or GSTT1 deletion were at higher risk of relapse and of shorter overall survival (Voso et al, Blood100:2703, 2002). The aim of the present study was to confirm or previous findings and to better define the role of GSTM1 and GSTT1 deletions in the context of a molecularly well-characterized cohort of AML-patients who were treated within two prospective clinical trials of the German-Austrian Acute Myeloid Leukemia Study Group (AMLSG) In addition to polymorphisms in GSTM1 and GSTT1, we were also interested in the role of the GSTP1(Ile105Val) polymorphism on outcome. Based on our previous work, we intended to show a 20% difference in the 3-year survival rate. Therefore the estimated power for our current study including 350 patients to confirm the previous findings was 98%. We studied 354 patients (pts) with AML, treated according to two AMLSG prospective multicenter clinical trials (HD98-A for pts with age &lt; 60 years, 254 pts; HD98-B for pts &gt; 60 yrs, 100 pts). The Ile105Val polymorphism of the GSTP1 gene (GSTP1*105) was analyzed by PCR-RFLP, while GSTM1 and GSTT1 homozygous deletions (GSTM1*0 and GSTT1*0) were studied using a multiplex PCR technique, including the housekeeping gene BCL-2 as internal control. GST genotypes were not associated to patients’ characteristics, including type of AML, cytogenetic risk group, presence of FLT3-ITD and abnormal laboratory parameters, and response to induction chemotherapy. GSTP1*105Val was associated with a significant better relapse-free survival using age-stratified test (adjusted HR, 0.7; 95% C.I., 0.51–0.98), while GSTT1 and GSTM1 genotypes had no impact on RFS. A Cox proportional hazard model on RFS including GST genotypes, LDH level, cytogenetic risk group, and FLT3-ITD mutation status showed that the GSTP1 genotype was a significant variable associated with a favourable outcome (n= 236 pts, HR, 0.62; 95% C.I., 0.44–0.88). Limiting the analysis to patients with a normal karyotype, the GSTP1 105Val allele was associated in the multivariable model on RFS with a 51% risk reduction (HR, 0.49; 95%C.I., 0.30–0.81). In conclusion, GSTP1*105Val seems to be of prognostic value, whereas the prognostic value of GSTM1 and GSTT1 reported in previously could not be confirmed.
Use of Post-Treatment Clinical Data To Predict Response to Decitabine.
Authors: Estey Elihu , Issa J.-P. , Garcia-Manero Guillermo , Faderl Stefan , Kantarjian Hagop 
Published: April 2020

Abstract
Best response of AML or MDS to decitabine (DAC) may require &gt; 3 courses. Identification of pts whose response to initial courses suggests they are unlikely to respond further to subsequent courses would allow such pts to receive other therapies more expeditiously. 104 pts, median age 69, received DAC (20 mg/m2 IV daily × 5 days every 5 weeks) for newly- diagnosed MDS (80, 76 IPSS int-2 or high) or AML (24) from 9/00-5/07. After 3 courses, 33 of the 104 were in CR, 6 were dead, and 32 had been removed from study, with the remaining 32 receiving more DAC. The probability of CR decreased to &lt; 10% on all courses after the 5th:
Course# Pts Receiving CR Deaths/Off Study Received Next Course after course Course 1 104 7 (7%) 2/15 80 2 80 16 (20%) 4/12 48 3 48 10 (21%) 0/6 32 4 32 5 (16%) 0/5 22 5 22 3 (14%) 0/3 16 6 16 1 (6%) 1/3 11 7–15 11 1 (9%) 1/9 0
Results were qualitatively similar for AML and MDS. We divided the 32 pts who received ≥ 4 courses according to whether after course 3 their:
marrow showed a response, defined as a p &lt; 0.05 difference between the pre-Rx and post course 3 blast count, platelet count increased &gt; 30, 000 if &lt; 100,000 pre-Rx, or neutrophil count increased &gt; 500 if &lt; 500 pre Rx; criteria 2 and 3 were adapted from those of the IWG. 8/14 pts with vs. 1/11 without a marrow response after course 3 entered CR on subsequent courses (p = .02, 6/10 vs. 1/10 considering only MDS).
The corresponding figure for platelet response/no response was 5/8 vs. 2/11 (3/5 vs. 2/9 for MDS). The median number of courses after the 3rd given pts with marrow response was 2 (range 1–10) vs. 3 (range 1–12) for pts without marrow response. Our data suggest that if CR is the goal, pts should not receive more than 5 courses of DAC and no more than 3 if marrow response has not occurred after the 3rd.
The Bone Resorption Marker Serum-ICTP and the Circulating Proteasome Levels Separate ISS-Stages 1–3 and Are the Most Powerful Prognostic Factors for Overall Survival in Newly Diagnosed Symptomatic Multiple Myeloma.
Authors: Jakob Christian , Liebisch Peter , Egerer Karl , Sterz Jan , Kaiser Martin , Rötzer Susanne , Heider Ulrike , Rademacher Jessica , Fleissner Claudia , Kloetzel Peter M. , Terpos Evangelos , Sezer Orhan 
Published: April 2020

Abstract
Prognostic markers are important to identify high-risk patients in Multiple Myeloma (MM). Several prognostic models have been published, including parameters of tumor burden and cytogenetics. Recently the International Staging System (ISS) for Multiple Myeloma, which includes beta2-microglobulin (beta2-MG) and albumin, has been introduced for newly diagnosed patients with symptomatic MM. We previously reported about the prognostic significance of circulating proteasome levels (cProteasomes) in MM patients (Jakob et al., Blood 2007). In the present study we investigated the prognostic relevance of the bone resorption marker carboxy-terminal telopeptide of type-I collagen (ICTP) and cProteasome levels in comparison to the classical prognostic factors (beta2-MG), albumin, deletion 13q14 and type of chemotherapy (high-dose-therapy versus conventional dose chemotherapy) in 92 patients with newly diagnosed active MM. ICTP and cProteasome were significantly elevated parallel to ISS stages (P&lt;0.001). ICTP (cut off: normal value), cProteasome levels (cut off: median value) and a combined ICTP-cProteasome score (1: ICTP &lt; normal value and cProteasome &lt; median; 2: one of both parameters elevated; 3: both parameters elevated) were significant univariate prognostic factors for overall survival in active MM (P&lt;0.001, P=0.002 and P&lt;0.001, respectively). Survival rates at 5-yrs were 95%, 66% and 27% in the groups of patients with ICTP-cProteasome score 1, 2 and 3, respectively. In a further analysis ICTP alone and the combined ICTP-cProteasome score significantly separated each of the three ISS stages into two distinct groups with a better versus worse prognosis. In a multivariate Cox regression analysis, including beta2-MG, albumin, deletion 13q14, type of chemotherapy, cProteasome levels and ICTP, ICTP was the parameter with the strongest prognostic power (P&lt;0.001, hazard-ratio: 7.9) and cProteasomes (P=0.011, hazard-ratio: 3). Our study underlines that the activity of myeloma bone disease, as reflected by the collagen-I degradation product ICTP, has a major impact on the prognosis of symptomatic MM. This result is in line with published data showing a positive feedback between myeloma cells and osteoclasts, i.e. a vicious cycle. The inclusion of the bone resorption marker ICTP and cProteasome levels into multivariate models add substantial prognostic information on overall survival in newly diagnosed active MM.
In Vivo Evaluation of a BAFF Inhibitor AMG 523 Suggests Lack of Efficacy in Multiple Myeloma Tumor Models.
Authors: Xu Jiangchun , Leenheer Evy D. , Piasecki Julia , De Vries Anne-Renee V. , Vanderkerken Karin , Croucher Peter 
Published: April 2020

Abstract
B-cell activating factor (BAFF), a member of the TNF superfamily, has been reported to play a role in the growth and survival of B-cell malignancies. However, the effect of inhibiting BAFF in vivo on tumor growth has not been fully determined. Additionally, studies so far have not addressed relative contributions of BAFF vs. APRIL towards cancer growth as the inhibitors used, such as TACI-Ig and BCMA-Ig, inhibit both BAFF and APRIL ligands. In the present study, we examined the potential therapeutic utility of a BAFF inhibitor, AMG 523, for treating B-cell malignancies in murine multiple myeloma in vivo models. AMG 523 is a peptibody that is a potent BAFF antagonist as a result of its high affinity binding to BAFF (KD of 1 pM). It inhibits BAFF-mediated increases in human and mouse B cell proliferation (IC50 of 0.1 nM at 0.2 nM BAFF). To determine the in vivo efficacy of AMG 523 in myeloma cells, we used the 5T series of murine models of myeloma. The 5T murine models of myeloma are myelomas that arose spontaneously in an inbred substrain of C57 black mice (C57BL/KaLwRij substrain). Several of the 5T myeloma lines closely mimic myeloma disease in humans. AMG 523 was first evaluated in a 5TGM1 syngeneic in vivo myeloma model. 5TGM1 was injected into the C57BL/KaLwRij mice and mice were treated with 5 or 20mg/kg of AMG 523 three times a week for four weeks. Tumor burden was determined by measuring the serum level of IgG2b, a paraprotein secreted by 5TGM1 myeloma cells. AMG 523 decreased serum IgG2b levels at early time points (day 8 and 15) but not at late time points (day 22 and 27) compared to the control. AMG 523 was also tested in two bone microenvironment dependent murine myeloma in vivo models: 5T33MMvv and 5T2MM. Both 5T33MMvv and 5T2MM cell lines express TACI receptor but not BCMA. BAFF increased survival of 5T2MM cells ex vivo, but inhibiting the activity of BAFF in vivo with AMG 523 had no effect on tumor burden, measured by proportion of bone marrow occupied by 5TMM cells, in both 5T33MMvv and 5T2MM models. Taken together, our data suggests that the BAFF inhibitor AMG 523 lacks efficacy in murine multiple myeloma in vivo tumor models.
A Novel Notch Pathway Inhibitor Blocks Osteoclast Activity and Synergistically Induces Apoptosis with the Proteasome Inhibitor Bortezomib in Multiple Myeloma Cells.
Authors: Jundt Franziska , Schwarzer Rolf , Kaiser Martin , Acikgoez Oezlem , Heider Ulrike , Mathas Stephan , Preissner Robert , Sezer Orhan , Dorken Bernd 
Published: April 2020

Abstract
Notch pathway inhibition in multiple myeloma (MM) cells is a promising new therapeutic approach since it controls myeloma cell growth as we previously demonstrated (Blood. 2004; 103:3511–3515). Notch signaling is involved in the tight interactions between myeloma cells and the bone marrow microenvironment and induces tumor cell growth in MM. We provided evidence that Notch receptors are expressed on MM cells and that Notch ligands on MM and bone marrow stromal cells activate Notch signaling through homotypic as well as heterotypic interactions in MM cells. In this study, we analyzed whether Notch signaling might be activated in osteoclasts, which express Notch receptors but not the ligands. To that end, we co-cultured MM cells and human osteoclasts, which were generated from mononuclear hematopoietic precursors of healthy donors using in vitro RANKL/M-CSF stimulation. Co-cultivation specifically activated Notch in osteoclasts and induced osteoclast activity as measured by mRNA expression of the tartrate-resistant acid phosphatase. The novel Notch pathway inhibitor, so called γ-secretase inhibitor 1 (GSI1), that we recently identified by structural comparison of known inhibitors with unknown compounds by data bank screening, specifically inhibited Notch signaling in osteoclasts and blocked their activity in this co-cultivation assay. In addition, GSI1 induced apoptosis in osteoclasts in higher concentrations. We suggest from our data that GSI treatment controls MM cell growth and concomitantly aberrant osteoclast activity in vitro and possibly in vivo, that is under current investigation. We further hypothesized that GSI1 can be combined with the proteasome inhibitor bortezomib, which has been known to have in vitro and in vivo activity against MM. We evaluated the activity of the combination of GSI1 and bortezomib against MM cell growth and survival. Proliferation of MM cell lines treated with GSI, bortezomib and their combination was determined by CellTiter-Glo® luminescent cell viability assay. AnnexinV-FITC/PI staining and cleaved poly (ADP-ribose) polymerase (PARP) staining were used to determine the degree of apoptosis. Although treatment of MM cell lines (OPM2, LP1) with either drug alone significantly inhibited proliferation and induced apoptosis with concentrations of GSI1 (30–60 μM) and bortezomib (1–4 nM), the combination resulted in synergistic inhibition of cell growth and survival. Our data suggest that combination of GSI1 and bortezomib is a rational novel treatment option in MM that simultaneously targets different proliferative and anti-apoptotic pathways. Whether this combination might also have synergistic activity against aberrant osteoclast activity in MM will be further investigated.
Inducible Expression of Leukemia-Associated Shp2 (Ptpn11) Affects Multiple Stages of Hematopoiesis and Causes a Fatal Myeloproliferative Disorder (MPD) in Mice
Authors: Chan Gordon , Kalaitzidis Demetrios , Mohi M. Golam , Yang Wentian , Kutok Jeffery , Neel Benjamin 
Published: April 2020

Abstract
Germ-line mutations in ptpn11, which encodes the protein tyrosine phosphatase Shp2, cause ∼50% of Noonan Syndrome (NS), which is associated with an increased risk of juvenile myelomonocytic leukemia (JMML). Somatic Shp2 mutations are found in ∼35% of sporadic JMML; nearly all other cases have either activating Ras mutations or homozygous Nf1 deficiency. Shp2 mutations are also found at lower incidence in acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), and B-cell acute lymphoblastic leukemia (B-ALL). NS and leukemia-associated Shp2 mutations can affect the same residue, but result in different substitutions. We showed previously that the leukemia-associated mutants E76K or D61Y, but not wild type (WT), Shp2 transform bone marrow (BM) or fetal liver cells. Transplantation of E76K- or D61Y-transduced BM evoked invasive MPD in ∼60% of recipients, with most dying at 6–7 months. In this model, mutant Shp2 is expressed under retroviral promoter control and the phenotype is variable and incompletely penetrant. To assess the effect of expressing a leukemogenic Shp2 allele under endogenous promoter control, we generated knock-in mice that can conditionally express Shp2D61Y (LSL-Shp2D61Y). Global expression of the D61Y heterozygous allele was embryonic lethal. Post-natal expression of D61Y, induced by treating Mx-1 Cre; LSL-Shp2D61Y mice with pI-pC, evoked fatal MPD with ∼50% mice dying within 5–7 months. Mutant mice showed a marked increase in WBC, expansion of the Gr-1+/Mac-1+population in BM and spleen, and histopathological evidence of infiltrating MPD. Unlike in our retroviral transduction/transplant model (and similar to JMML patients), most induced LSL-Shp2D61Y mice were also anemic. Mutant mice exhibited a marked depletion of quiescent LSK (Lin−Sca1+cKit+) cells in the BM, with a concomitant increase in LSK cells in the spleen. Interestingly, Shp2 mutant-expressing BM failed to promote long term reconstitution in BM transplant assays. Cells from mutant spleens did have some long term multi-lineage reconstitution activity, but the level was substantially less than predicted by their LSK content. Moreover, no transplant recipients develop MPD. In contrast to these effects on the stem cell compartment, D61Y directly allowed cytokine-independent differentiation of CMP and GMP in vitro. Macrophages from mutant mice showed enhanced GM-CSF-mediated proliferation and ERK activation. Our results showed that expression of leukemia-associated Shp2 at endogenous levels is sufficient to evoke MPD and has cell type-specific effect on different stages of hematopoiesis.
Characterization of del20q in Peripheral Blood of MPD Patients Using Copy Number Analysis and High Resolution Oligonucleotide CGH Array
Authors: Schaub Franz , Tiedt Ralph , Hermouet Sylvie , Girodon François , Kralovics Robert , Tichelli André , Skoda Radek C. 
Published: April 2020

Abstract
Previous studies showed that deletions of the long arm of chromosome 20 (del20q) are the most common chromosomal aberration in myeloproliferative disorders (MPD) and can be found in up to 10% of patients with polycythemia vera (PV). We screened 447 MPD patients (177 PV, 228 essential thrombocythemia (ET), 42 primary myelofibrosis (PMF)) for loss of gene copy number in the del20q region. Granulocyte DNA was assayed using a quantitative PCR assay for two different exons of the L3MBTL gene, which is located in the common deleted region. Since deletions can only be detected if the majority of granulocytes have lost a copy of L3MBTL, this method selects for del20q events that provided a selective advantage for the clone. Samples that showed a decrease in copy number in the initial screen were validated using primers in neighbouring genes. We found del20q in peripheral blood of 29/447 patients (6%), including 9/177 PV (5%), 12/228 ET (5%), and 8/42 PMF (19%). The observed frequency of del20q in peripheral blood is lower than reported in previous studies for bone marrow analysis. The patients positive for del20q in this screen were chosen for mapping of the deleted region by high density oligonucleotide comparative genomic hybridisation (CGH) array. A custom chip was designed with 94’000 probes from chromosome 20q, with an average resolution of 500bp. We have so far analyzed 6 patients and confirmed del20q by CGH in all of them. The deleted regions in these patients overlapped with the published common deleted region. We will use CGH to map the remaining 23 patients. Since the majority of the del20q positive patients also carried the Jak2-V617F mutation, we performed clonogenic assays and analyzed single colonies. In one patient all 94 colonies were double positive for del20q and JAK2-V617F. In a second patient 54 colonies were double positive and 10 colonies only carried del20q, but not JAK2-V617F, indicating that del20q occurred before the acquisition of JAK2-V617F.
Association between Polymorphic Variation at the IL5RA Locus and the Severity of FIP1L1-PDGFRA Positive Chronic Eosinophilic Leukaemia
Authors: Burgstaller Sonja , Kreil Sebastian , Waghorn Katherine , Metzgeroth Georgia , Preudhomme Claude , Zoi Katerina , White Helen , Cilloni Daniela , Zoi Christine , Brito-Babapulle Finella , Walz Christoph , Reiter Andreas , Cross Nicholas C.P. 
Published: April 2020

Abstract
FIP1L1-PDGFRA is a cytogenetically cryptic fusion gene seen in a minority of patients with chronic eosinophilic leukaemia (CEL). Observations in mice have suggested that FIP1L1-PDGFRA alone is insufficient to induce all features of the human disease, and that a secondary event associated with IL-5 overexpression may be required for the full phenotype. In this study we have investigated the hypothesis that constitutional genetic variation in IL-5 signalling may be associated with the development or severity of FIP1L1-PDGFRA positive CEL in humans. We genotyped six single nucleotide polymorphisms (SNP) within or close to the IL5RA or IL5 genes in 82 European patients with FIP1L1-PDGFRA positive CEL plus, as controls, healthy individuals (n=100), patients with FIP1L1-PDGFRA negative eosinophilia (n=100) or patients with chronic myeloid leukaemia (n=100). We found no association between SNP allele frequency between FIP1L1-PDGFRA positive and control cases. However for FIP1L1-PDGFRA cases, we found an association between the genotype at rs4054760, a SNP in the 5′UTR of IL5RA, and peripheral blood eosinophil count (P=0.026) as well as the presence or absence of tissue infiltration (P=0.032). Although these associations fell below the level of significance once corrected for multiple testing, no such association was seen in FIP1L1-PDGFRA negative cases and no difference in allele frequencies for rs4054760 was seen in control populations across Europe. Furthermore, in an analysis of 128 patients with CML, there was a trend towards lower expression of IL5RA expression in cases with a c/t or t/t genotype compared to those who were c/c (P=0.08). These data suggest that the constitutional IL5RA genotype may be associated with the severity of FIP1L1-PDGFRA disease.
Comprehensive Highly Multiplexed Single-Tube Liquid Bead Microarray Assay for Constitutively Activated Tyrosine Kinases in Chronic Myeloproliferative Disorders.
Authors: Roberts Mark J. , Zhou Yening , Miller Kenneth B. , Pihan German A. 
Published: April 2020

Abstract
Chronic Myeloproliferative disorders (CMPDs) are malignancies characterized by excessive proliferation of one or more of the myeloid lineages driven by constitutively activated tyrosine kinases (CATKs). CATKs are increasingly serving as targets for cancer-specific therapy and newer and more potent TK inhibitors are becoming available for clinical use. CATKs in CMPDs may result from karyotypically evident or cryptic chromosomal translocations leading to TK gene fusions or from karyotypically silent gain-of-function point mutations. More than ten CATK genes have been shown to result in CMPDs by imparting a demonstrable positive growth drive. CATK inhibitors important in the treatment of CMPDs have different inhibitory profiles, making it critical to identify the specific mutation involved in patients receiving targeted therapy. Of these TK genes, ABL, PDGFRB, PDGFRA, FGFR1, ETV6, JAK2 and MPL are the most frequently involved. Since most of these TK genes can participate in chromosome translocations with a large number of partner genes, the resulting number of possible CATKs is rather large, making detection and identification a challenging task. To date, two classic (B2A2 and B3A2) and seven less common fusions of the ABL and BCR genes have been described. In addition, over twenty non-BCR-ABL mutations have been shown to have pathogenetic relevance in CMPDs. We have developed a simple, high-throughput assay with a fast turnaround time that solves the complex problem of detecting and identifying multiple translocations and point mutations. Two steps are involved in this expandable assay. The first is a multiplex RT-PCR reaction that amplifies fusion transcripts and point mutations described in CMPDs from peripheral blood or bone marrow. This PCR reaction employs internal specific primers targeted to each of eight BCR-ABL mutations (E1A2, E1A3, B2A2, B3A2, E6A2, E8A2, E15A2, E19A2) eight non-BCR-ABL mutations (FIP1L1-PDGFRA, BCR-PDGFRA, BCR-FGFR1, MYO18A-FGFR1, CDK5RAP2-PDGFRA, ETV6-PDGFRB, RABAPTIN-5-PDGFRB, H4-PDGFRB) and two gain of function mutations (JAK2V617F and MPLW515L) and a biotinylated common primer set to increase amplification efficiency. The second is a liquid bead microarray (Luminex) which uses optically encoded beads to detect probes specific for multiple known translocations in a single tube. To date, we have selectively detected four BCR-ABL and three non-BCR-ABL mutations from cloned patient samples and the cell lines SupB15 (E1A2 p190 BCR-ABL fusion), K562 (B3A2 p210 BCR-ABL fusion) and BV173 (B2A2 p210 BCR-ABL fusion). Dilution studies using these cell lines demonstrate detection down to between 1/1000 and 1/100000 cell equivalents without significant cross-talk.
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