bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Cell Expression of PML-RARa at Diagnosis Has a Marginal Correlation with Prognosis in Patients with Acute Promyelocytic Leukemia (APL).
Authors: Fernández Carina , Chillón Carmen , Blázquez Lorena , Ramos F. , Fernández-Calvo J. , Rayón C. , Balanzategui A. , Martín-jiménez P. , Alcoceba M. , Sarasquete M. E. , Armellini A. , García-Sanz R. , González M. , Miguel J.F. San 
Published: April 2020

Abstract
Acute promyelocytic leukaemia (APL) is characterised by the t(15;17) leading to the fusion of the PML gene with the retinoic acid receptor a (RARa) gene. APL is associated with favourable prognosis, however, approximately 15–20% of patients ultimately relapse. With regards to pre-treatment characteristics, is now clear that long-established prognostic factors such as age and leukocyte count have a major impact on outcome. However, none of these parameters is robust enough, in order to adjust treatments strategies according to the risk of relapse. The quantification of PML-RARa fusion gene transcripts (leukemia-specific chimeric mRNA) can add important prognostic information useful for risk group stratification. However, few clinical studies have been carried out and there are discrepancies concerning the prognostic significance of the quantification of fusion gene transcripts of newly diagnosed patients. In this study we tested the PML- RARa fusion gene transcripts level in 126 newly diagnosed patients with t (15;17) in order to determine their correlation with disease characteristics at presentation and to identify a subset of patients at high risk of relapse. The presence of the PML- RARa fusion gene transcripts was analysed from 1mg of RNA by real-time quantitative RT-PCR (RQ-PCR) based on TaqMan probes and the 7700 Sequence Detector, performed according to the “Europe Against Cancer Program” (Leukemia2003, 17:2323; 2318–2357).
Results: In 121 out of 126 patients, evaluable data were obtained: 73 (60,3%) cases had bcr-1, 4 (3,3%) cases had bcr-2 and 44 (36,4%) cases had bcr-3 fusion types. The expression of the fusion gene was reported as the PML- RARa normalized quantities (NQ, number of PML- RARa copies per 1x104 ABL copies as gene control). The median of NQ was 3030 and the expression was highly variable (range of 826 to 9605). The NQ was not significantly associated with disease characteristic at diagnosis, such as age, percentage of blasts cells, fusion types and platelet counts. Nevertheless, patients with NQ above 3000 had lower white cell counts (7,4±12,1 vs 24,2±39,3x109/L, p=0,03). Therefore, the fusion transcript copy number per leukemic cell was higher in patients with lower leukocyte count. Moreover, in survival analysis, the NQ did not correlate with disease free survival or overall survival.
Conclusions: Our data confirms the wide range of differences in expression activity between individual patients. The NQ did not correlate with the clinical and biological disease characteristics except for white cell counts. Neither did it correlate with principal prognostic parameters (response and survival).
Genetic Instability in Remission Hematopoiesis Is Different from That in AML Blasts: A New Diagnostic Tool to Study Biology of AML.
Authors: Reichle Albrecht , Panzer Birgit , Goetz Gerlinde , Raffer Michael , Zaiss Matthias , Andreesen Reinhard , Wild Peter , Hofstaedter Ferdinand , Dietmaier Wolfgang 
Published: April 2020

Abstract
Recently we have shown that aberrations, ie microsatellite instability (MSI) and allelic imbalance (AI) in positively selected CD34+ cells from leukapheresis products collected during first complete remission of de novo AML following double induction or consolidation according to AMLCG protocol may be predictive for relapse-free survival (RFS). Here we present data of an extended patient population treated on the same protocol. We compared MSI/AI pattern in CD34+ cells (n=60) with the corresponding pattern in AML blasts at diagnosis (n=60) or at relapse (n=12), and compared the AI/MSI pattern in CD34+ cells with those observed in the corresponding unselected bone marrow aspirates in first CR (n=50). The following loci were tested: D7S486, D7S525, D8S559, TP53, D11S1356, D2S123, APC, MfD5. Minimal residual disease (MRD) in the tested remission specimen (FACS analysis) was &lt;0.1%. MSI and/or AI at diagnosis could be detected in few cases (10 of 60, 17%), whereas CD34+ cells were tested positive in 25 of 60 cases (42%), and in 5 of 12 relapses studied (42%). Analysis of AI and MSI in DNA of bone marrow aspirates could not adequately confirm MSI and/or AI detectable in the corresponding CD34+ cells (n=3 cases, 12%). In 15 cases MSI and/or AI was observed in CD34+ cells, but not in AML blasts at diagnosis, in 5 cases in blasts at diagnosis, but not in CD34+ cells, in 4 cases in AML blasts at diagnosis and in CD34+ cells. Identical AI/MSI pattern were found in CD34+ cells and in AML blasts at relapse (n=2), as well as in AML blasts at diagnosis and relapse (n=1). According to the definition of genetic instability (GIN: MSI or &gt;2 AI at any locus, AI at APC locus), GIN in CD34+ cells was predictive for unfavorable RFS (p=0.001). In conclusion, detection of AI/MSI in CD34+ cells of remission hematopoiesis represents a new important diagnostic tool to study biology of AML besides the detection of MRD. For the first time we could demonstrate, that the same AIs or MSIs detectable in CD34+ cells, but not in AML blasts at diagnosis may occur in AML blasts at relapse.
High Levels of Soluble HLA-I in Patients with Acute Myeloid Leukemia and Myelodysplastic Syndrome: Relevance to Clinical Behavior and b2M Levels.
Authors: Albitar Maher , Johnson Marcy , Do Kim-Ann , Day Amanda , Jilani Iman , Pierce Sherry , Estey Elihu , Kantarjian Hagop , Keating Michael , Verstovsek Srdan , O’Brien Susan , Giles Francis J. 
Published: April 2020

Abstract
Human leukocyte antigen-class I (HLA-I) molecules are membrane-associated proteins that contain two separate polypeptide chains: an alpha heavy chain and a beta chain. The beta chain is the b-2 microglobulin (b2-M). Significant levels of free soluble HLA-I (sHLA-I) have been detected in serum and studied as a marker for immunomodulation in patients with infection and after transplant. Little is known about the role of sHLA-I as a tumor marker, despite the documentation of b2-M as a tumor marker. We measured the levels of sHLA-I and b2-M in the plasma of 205 patients with acute myeloid leukemia (AML) and 95 patients with myelodysplastic syndrome (MDS) and assessed the value of both markers in predicting clinical behavior. Both sHLA-I and b2-M were strong predictors of response to therapy (P = 0.03 and P = 0.001, respectively), overall survival (both P &lt;0.0001), and remission duration (both P &lt;0.001). The Multivariate Cox proportional hazards model incorporating cytogenetics, b2-M, and sHLA-I demonstrated that only b2-M was an independent predictor of survival. In patients with MDS, b2-M but not sHLA-I correlated with response to therapy, overall survival, and response duration. These data not only establish the role of sHLA-I as a tumor marker in AML, but also suggest that MDS disease is significantly different from AML. Because sHLA-I has been reported to be an immunomodulator that inhibits the cytotoxic effects of T-lymphocytes, its role as an immunomodulator in patients with MDS needs further investigation.
Figure Figure
Clinical Relevance of Circulating Myeloperoxidase (MPO) in Acute Myeloid Leukemia (AML) and Myelodysplastic Syndrome (MDS).
Authors: Jilani Iman , Vincenti Ted , Faraji Homan , Giles Francis J. , Estey Elihu , Kantarjian Hagop M. , Albitar Maher 
Published: April 2020

Abstract
MPO, a hemoprotein expressed in polymorphonuclear neutrophils and their precursors, is an important component of the antimicrobial activity of phagocytes and is associated with inflammatory response. Cellular expression levels of MPO, assessed by cytochemical staining, are used extensively for the diagnosis and subtyping of various leukemias. However, the association of circulating (plasma or serum) MPO levels with hematologic values and clinical outcomes of AML and MDS has not been reported. We investigated whether MPO, presumably released from leukemic cells, can be detected in the plasma of patients with AML and MDS. We further assessed the association of plasma MPO levels with hematologic characteristics and outcomes. 144 patients with AML, 28 patients with MDS, and 100 control subjects were included in the analysis. Patients with AML (median 19.3, range 1.0–9514.7 ng/mL) and MDS (median 13.6, range 1.0–3021.9 ng/mL) had significantly higher plasma MPO levels than control subjects (median, 4.9; range, 3.5–20.6 ng/mL). Because MPO levels and overall survival did not differ significantly between AML and MDS patients, the 2 groups were considered together in subsequent analyses. MPO levels were inversely correlated with overall survival (Cox regression analysis, P&lt;0.001). In the multivariate model, the inverse correlation between MPO levels and survival was independent of cytogenetics, performance status, anticident hematologic disease (AHD), age, and diagnosis (AML vs MDS). Response to therapy in AML and MDS patients was not associated with plasma MPO levels. However, responders with high MPO levels had shorter remission duration (CRD), although this difference did not reach statistical significance (P = 0.07). As expected, MPO levels correlated with the level of differentiation of leukemic cells (FAB subclassification): patients with M0 and M6 had lower MPO levels than those with M2, M3, or M4 (P = 0.0001). MPO levels also correlated positively with white cell count, beta2-microglobulin, monocyte, LDH, BUN, and creatinine levels. These data indicate that plasma MPO levels are elevated in AML and MDS and may be useful as a prognostic factor. Our findings also raise the question of how the proinflammatory properties of MPO might affect the clinical course of these diseases.
CALM-AF10 Positive T-ALLs Show a Pattern of Expression Similar to MLL-Translocated Acute Leukemias.
Authors: Delabesse Eric , Dik Wim A. , Brahim Wajih , Braun Charlene , Asnafi Vahid , Van Dongen Jacques J.M. , Langerak Anton W. , Macintyre Elizabeth A. 
Published: April 2020

Abstract
The t(10;11) translocation is recurrent in T-ALL and AML. The AF10 gene on chromosome 10 is rearranged either with MLL or CALM located on chromosome 11. CALM-AF10 fusion gene is found in T-ALLs in immature (IM) and TCRγδ-expressing (TCRGD+) T-ALLs. We compared 6 CALM-AF10+ T-ALLs cases (4 IM, 2 TCRGD+) to 17 CALM-AF10 negative T-ALLs cases (14 IM, 3 TCRGD+) using Affymetrix U133A microarrays.
44 genes were significantly overexpressed in CALM-AF10+ T-ALLs, the most significant being HOXA9, a homeobox gene overexpressed in MLL-translocated acute leukemias (MLL-t AL), BMI1, a polycomb family member whose function in regulation of HOX genes expression is opposite to Trithorax genes (whose MLL belongs), SOX4, a frequent insertion site in retroviral-induced leukemogenesis, SFRS6 and COMMD3 (p≤0.001). Only two other HOX genes, HOXA5 and HOXA10, were significantly increased. 89 genes were significantly underexpressed in CALM-AF10+ T-ALLs, the most significant being GGH, ARL6IP4, NBS1, OGFR and TUBB (p≤0.001).
An independent analysis of the expression of HOXA5, HOXA9, HOXA10 and BMI1 genes was done by quantitative RT-PCR in 10 CALM-AF10+ T-ALLs and 27 CALM-AF10 negative T-ALLs. These were compared to 19 MLL-translocated acute leukemias (2 MLL-AF10, 5 MLL-AF4, 3 MLL-AF6, 5 MLL-AF9, 3 MLL-ELL and 1 MLL-ENL), since HOXA9 overexpression had been previously associated with MLL-t AL.
HOXA5, HOXA9 and HOXA10 expression were higher in CALM-AF10+ T-ALLs than in CALM-AF10 negative T-ALLs (p&lt;0.001), confirming the microarray results. HOXA5 and HOXA9 expressions in CALM-AF10+ T-ALLs were similar to those detected in MLL-t AL and lower for HOXA10 in CALM-AF10+ T-ALLs as compared to the values of MLL-t AL (p=0.008). BMI1 expression in CALM-AF10+ T-ALLs was significantly higher than in CALM-AF10 negative T-ALLs and MLL-t AL (p&lt;0.001).
Additionally, MEIS1 expression was determined as this gene was associated in MLL-t AL with the overexpression of HOXA9. As for BMI1, MEIS1 expression was significantly higher in CALM-AF10+ T-ALLs compared to CALM-AF10 negative T-ALLs and MLL-t AL (p&lt;0.001 and p=0.019, respectively).
In summary, we demonstrated here the association between CALM-AF10 in T-ALLs and overexpression of HOXA5, HOXA9, HOXA10, BMI1 and MEIS1 genes. Overexpression of BMI1 is restricted to CALM-AF10+ T-ALLs. Although no obvious similarities are apparent between MLL and CALM proteins, the activation of HOXA and MEIS1 genes represent a highly recurrent pattern of expression in CALM-AF10+ T-ALLs and MLL-t AL. Consequently, the leukemias resulting in the activation of HOXA9 (MLL-t AL, CALM-AF10+ AL, NUP98-HOXA9 AML) should be seen as an independent group of acute leukemias and may benefit from common therapeutic protocols.
Identification of 41 Novel Promoter-Associated CpG Islands Methylated in Leukemias.
Authors: Jelinek Jaroslav , Mannari Rajan , Issa Jean-Pierre 
Published: April 2020

Abstract
DNA methylation within promoter-associated CpG islands is a well-recognized mechanism of gene silencing and plays an important role in the development of malignancies. CpG dinucleotides in human DNA are methylated at 5′-cytosine with the exception of areas with dense concentration of CpGs (CpG islands) located in gene promoter regions. In cancer cells, methylation of CpG islands in promoter regions of tumor suppressor genes is a frequent epigenetic change with a gene-silencing effect analogous to inactivating mutations. Methylation profiling can identify biologically and clinically distinct tumor subgroups by mapping the methylation status of multiple genes, and reports in AML and ALL suggest associations between methylation and poor prognosis. Identification of methylated CpG islands can shed new light on the biology of leukemia. We used Methylated CpG Island Amplification coupled with Representative Difference Analysis (MCA-RDA) as a genome-wide screen for promoter-associated CpG islands methylated in leukemic and/or myeloproliferative cell lines and primary malignant cells, but unmethylated in blood cells from normal controls. We identified 51 unique promoter-associated CpG islands in 321 sequenced clones recovered by MCA-RDA. Forty-one CpG islands belonged to known genes, and 10 to annotated mRNAs. Of the genes with known function, 8 are involved in signaling, 7 in transcription, 3 in dephosphorylation, 2 in oxido-reductive processes, 2 in NO synthesis, 2 in adhesion, 2 in solute transport, and 2 in DNA replication. Seven out of the 51 genes were previously reported as methylated in cancer or leukemia (CDH13, HLA-B, HLA-C, PGR, SCGB3A1, SLC26A4, TERT), thus validating the MCA-RDA approach. Of the 41 new hypermethylated CpG islands recovered, 20 corresponded to genes of known function. Published data infer an association with cancer for 10 of these genes (CTDSPL, ECGF1, EDG4, FOXD2, NOR1, NOS3, OLIG2, SLC16A1, TLE1, WNT5B), and no reports were found for the other 10 genes (CNR1, FADS, FBXW3, FGD1, NPM2, P518, PDE4DIP, SNCB, TCEA3, VENTX2). To further validate our findings we are assessing the methylation status of these genes by bisulfite pyrosequencing. Analyses of the bone marrow samples from AML, ALL, CML and MDS patients are ongoing. Our preliminary data confirm methylation of H-cadherin precursor (CDH13), progesterone receptor (PGR) in AML and ALL and cannabinoid receptor 1 (CNR1) in ALL (Table). In conclusion, MCA-RDA identified methylation of 41 new and 10 previously reported promoter-associated CpG islands in leukemia. Functional studies of these may shed new light on the biology of leukemias, and these genes may be useful for methylation profiling of leukemias for prognosis and response to treatment.
Promoter CpG Island Methylation
Gene AML ALL Methylation levels over 10% for CDH13 and PGR and over 25% for CNR1 were scored as positive. CDH13 5/23 22% 12/19 63% PGR A Isoform 11/22 50% 12/18 67% PGR B Isoform 17/24 71% 5/13 38% CNR1 0/24 0% 3/18 17%
Inhibition of Retinoic Acid Receptor Signaling by SKI in Acute Myeloid Leukemia.
Authors: Ritter Markus , Samuelsson Magnus , Hartmann Oliver , Burchert Andreas , Kim Theo D. , Thiede Christian , Hayman Micheal J. , Ueki Nobuhide , Eilers Martin , Neubauer Andreas 
Published: April 2020

Abstract
Objectives: Acute myeloid leukemia (AML) with monosomy 7 or deletion of the long arm of chromosome 7 (-7/7q-) is a leukemia with a poor outcome. Microarray analysis resulted in a gene expression profile specific for this subset of AML and showed SKI, a nuclear Co-repressor protein, being strongly upregulated. SKI is described to be a transforming gene, which inhibits TGF-beta-, SMAD- and retinoic acid (RA)-signaling.
Materials, methods and patients: Gene expression profiling was performed using cDNA microarrays (detecting 4608 genes). Gene expression pattern of 20 AML (9 pts. with -7/7q- and 11 pts. with normal karyotype) patients was compared to 23 healthy bone marrow donors. Expression data of interestings genes were confirmed by real-time PCR in samples of 111 AML patients. In cell culture experiments HL60 and U937 leukemic cell lines were used to investigate the influence of the identified oncogene Ski on differentiating properties. In a reporter gene assay retinoic acid response element (RARE) was transfected in QT6 cells. U937 were retrovirally transfected with Ski wild type and Ski mutant.
Results: Microarray analysis in AML (-7/7q-), revealed SKI being upregulated in AML. The group of patients with a high SKI expression level had a poorer overall survival when compared to the low expressers (p = 0.0279). Real-time PCR analysis comparing 111 AML samples to healthy CD34+ haematopoietic stem cells revealed, that SKI levels were highest in AML with -7/7q-, moderately elevated in most other karyotypes except AML with translocations involving retinoic acid receptor alpha (RARalpha). This suggests that Ski might interfere with RARalpha function in leukemia. Using reporter gene assays we could show that expression of wild-type, but not mutated SKI blocked RA induced differentiation. SKI was down regulated in HL60 leukemia cell line when differentiation was induced by ATRA and or valproic acid.
Conclusion: Upregulation of SKI has prognostic significance in AML. Inhibition of RA signaling may be more often implicated in pathogenesis of AML than anticipated so far. The Co-repressor of histone deacetylases (HDACs) Ski might therefore become a molecular target for the induction of differentiation in the treatment of AML.
Treosulfan-Based Myeloablative Conditioning Prior to Allogeneic Hematopoietic Cell Transplantation Is Associated with Reduced Toxicity without Increased Risk of Relapse.
Authors: Giebel Sebastian , Wojnar Jerzy , Krawczyk-Kulis Malgorzata , Wylezol Iwona , Markiewicz Miroslaw , Kruzel Tomasz , Holowiecki Jerzy 
Published: April 2020

Abstract
The goal of this study was to evaluate toxicity and efficacy of allogeneic hematopoietic cell transplantation (alloHCT) with Treosulfan (alkylyting agent, soluble Busulfan-derivative)-based conditioning as a preparative regimen. The outcome of 27 patients (CML-15, AML-9, ALL-1, SAA-2) given Treosulfan 3x14 g/m2 + Fludarabine 5x30 mg/m2 (n=15) or cyclofosfamide (CTX) 120 mg/kg (n=12) was compared with that of 146 patients treated with Busulfan 16 mg/kg + CTX 120 mg/kg between 2000–2004. In case of unrelated donor (URD)-HCT patients were additionally given anti-thymocyte globulin. GVHD prophylaxis consisted of cyclosporin A and short-course Methotrexate. The indications for alloHSCT were comparable for both subgroups. The patients age was 35(14–56)y and 30(14–55)y, the proportion of URD-HCT was 55% and 45%, peripheral blood was used as a source of stem cells in 33% and 11% of cases, and the proportion of patients with advanced disease (AML and ALL &gt;CR1; CML &gt; CP1) equaled 26% and 19%, respectively.
The cumulative incidence of non-relapse mortality at one year was 8% for the Treosulfan (EBV-LPD n=1, cerebral hemorrhage n=1) and 32% for the Busulfan group (p=0.11). Grade II-IV neutropenic infections occurred in 5% and 13% (p=NS) and grade II-IV mucositis in 9% and 61% of patients (p&lt;0.001), respectively. Other serious adverse events were infrequent. Both subgroups did not differ in terms of time to neutrophil and platelet recovery, as well as for need of RBC and platelet transfusions. Median hospital stay since the date of alloHCT was shorter after Treosulfan- compared to Busulfan-based conditioning (31 (21–55) d. vs. 40.5 (21–153) d., p&lt;0.001).
The overall survival and disease-free survival at one year equaled 92% vs. 67% (p=0.11), and 92% vs. 66% (p=0.09) for the Treosulfan group and the Busulfan group, respectively. None of the patients experienced hematologic relapse among Treosulfan- compared to 3% among Busulfan-treated alloHCT recipients. However, in two CML patients given Treosulfan-based regimen, the immunosuppression taper and interferon or imatinib therapy was neccessary to establish complete chimera. We conclude that Treosulfan + Fludarabine (or CTX) +/− ATG conditioning regimen is characterized by reduced toxicity resulting in shorter hospital stay and low transplat-related mortality. The pattern of hematopoietic recovery is similar to that of Busulfan + CTX indicating myeloablative character of the Treosulfan-based regimen. The risk of relapse is low and comparable for both kinds of treatment.