bluuebunny/crossref_metadata_2025_split huggingface dataset:
Impact of Different Postremission Strategies in Younger Adults with Acute Myeloid Leukemia and Normal Karyotype Exhibiting a CEBPA Mutation.
Authors: Schlenk Richard F. , Corbacioglu Andrea , Fröhling Stefan , Du Juan , Späth Daniela , Schäfer Irina , Krauter Jürgen , Ganser Arnold , Döhner Hartmut , Döhner Konstanze ,   for the German-Austrian, AML Study Group (AMLSG)
Published: April 2020

Abstract
Recently, mutations in the CEBPA gene, encoding CCAAT/enhancer binding protein alpha have been identified in 10 to 15% of patients with acute myeloid leukemia (AML) exhibiting a normal karyotype and mutant CEBPA was shown to predict favorable outcome. However, the impact of different postremission strategies such as high-dose cytarabine based chemotherapy (chemo), autologous (auto-SCT) or allogeneic stem cell transplantation (allo-SCT) in subgroups defined by these molecular markers is under discussion. Therefore, we initiated a pooled data analysis on patients exhibiting a normal karyotype at diagnosis treated within the prospective treatment trials AML-2/95, AML-1/99, AMLHD93 and AMLHD98A.
All patients (age 16–60 years) received two cycles of induction therapy with standard dose cytarabine combined with etoposide and idarubicin. After a first consolidation therapy, patients were assigned to an allo-SCT if an HLA-identical sibling donor was available in all four trials. In the AML-2/95 and AMLHD93 trials all other patients were assigned to chemo whereas in the AML-1/99 and AMLHD98A trials patients were randomised between auto-SCT and chemo. All patients were analyzed for CEBPA-mutations in the N-terminal transactivation domains and the C-terminal basic region-leucine zipper domain by direct sequencing of genomic DNA.
Between 1993 and 2004 a total of n=872 patients exhibiting a normal karyotype had been registered. Actually in 413 patients results of CEBPA analyses are available and 54 (13%) exhibited a mutation (46% N-terminal, 15% C-terminal, 39% N- and C-terminal). Response to induction therapy was 85% (46/54) and 74% (259/352) in the CEBPA mutated group and the CEBPA wild-type group, respectively. RFS and OS were 59% and 38% (p=0.01) as well as 66% and 41% (p=0.002) in the CEBPA mutated group and the CEBPA wild-type group, respectively after a median follow-up of 42 months. The distribution of postremission therapy of the 46 patients in the CEBPA mutated group was n=22 Chemo, n=13 allo-tpl, n=9 auto-tpl, n=2 allo-tpl from a matched unrelated donor. Analysis based on the availability of a matched related donor revealed a significant better RFS (p=0.03) and OS (p=0.03) for patients with an matched related donor.
Conclusion: Although AML patients with normal karyotype and a CEBPA mutation comprises a group with favorable prognosis allogeneic transplantation from an HLA-matched family donor seems to improve long term outcome. Confirmation of these preliminary data on a larger series of patients are necessary to improve a risk-adapted decision on postremission strategies.
Protein Kinase CK2 Is Associated with Poor Clinical Outcome and Regulates the Expression of Anti-Apoptotic Proteins and PTEN Phosphorylation in Acute Myeloid Leukemia.
Authors: Cheong June-Won , Eom Ju In , Lee Jin Koo , Park Sun Young , Kim Yuri , Park In Hae , Kim Jin Seok , Min Yoo-Hong 
Published: April 2020

Abstract
We previously demonstrated that the phosphorylation of c-terminal regulatory domain of phosphatase and tensin homologue (PTEN) protein are frequently observed and significantly associated with poor clinical outcome in acute myeloid leukemia (AML). Phosphorylation of PTEN stabilizes the PTEN protein, but makes it less active towards its substrate, phosphatidylinositol 3,4,5-triphosphate, resulting in an antagonizing the apoptotic pathways. Protein kinase CK2 is implicated in many cellular processes including the cell proliferation, survival, and tumorigenesis. However, the biological significance of CK2 expression and its association with PTEN phosphorylation has not been evaluated. In this study, we evaluated the protein kinase CK2 activity/expression and PTEN phosphorylation simultaneously in leukemic blasts obtained from 59 patients with AML with normal karyotype. Western blot analysis revealed that PTEN phosphorylation was observed in 30 (50.9%) cases. CK2 expression was significantly associated with CK2 catalytic activity and PTEN phosphorylation. Levels of CK2 catalytic activity were significantly higher in AML cases with PTEN phosphorylation (p-PTEN-positive) compared with those without PTEN phosphorylation (p-PTEN-negative) (P&lt;0.05). Age, sex, white blood cell count, and complete remission rate were not different according to the CK2 expression. However, the overall survival was significantly shorter in AML cases with high CK2 expression (P&lt; 0.05). Treatment of p-PTEN-positive AML cells with apigenin, a specific inhibitor of CK2, abolished the phosphorylation of PTEN, as well as Ser473 phosphorylation of Akt/PKB, in a dose-dependent manner. CK2 overexpression induced an increase in the proportion of cells in the G(2)/M phase. Induced overexpression of CK2 in U937 leukemia cells led to PTEN phosphorylation, Ser473 Akt/PKB phosphorylation, in addition to an increased expression of Bcl-2, Bcl-xL, Mcl-1, Survivin, and XIAP proteins. Taken together, these results suggest that the phosphorylation of PTEN and expression of anti-apoptotic proteins were directly regulated by protein kinase CK2 in AML cells. We also demonstrate, for the first time, that CK2 is an independent prognostic variable in AML patients with normal karyotype. These findings additionally extend our understanding of the role of CK2 in AML, and suggest the CK2 molecule as a potential target for AML treatment.
NPM Mutations in Adult AML with Normal Karyotype: A Retrospective Study of the Acute Leukemia French Association (ALFA).
Authors: Boissel Nicolas , Renneville Aline , Biggio Valeria , Philippe Nathalie , Thomas Xavier , Cayuela Jean-Michel , Terre Christine , Tigaud Isabelle , Castaigne Sylvie , Raffoux Emmanuel , de Botton Stephane , Fenaux Pierre , Dombret Hervé , Preudhomme Claude 
Published: April 2020

Abstract
NPM mutation (NPMm) has been recently reported as the most frequent mutation in AML, especially in the presence of a normal karyotype. Association of NPMm with an increased complete remission (CR) rate has been suggested, but the long-term prognosis is not known. The abnormal mutated NPM protein shows an aberrant cytoplasmic localization that allows an immunohistochemical detection of NPM status. However, all mutations reported are insertion/deletion of exon 12 resulting in a global insertion of 4 nucleotides. We therefore developed a NPMm detection test based on DNA PCR amplification and fragment length analysis. As previously reported, we observed a higher frequency of NPMm in AML with normal karyotype (47%, 50/106) than in CBF AML (0%, 0/7) or in AML with poor risk cytogenetic (13%, 4/32). We further evaluated the clinical profile and the prognosis of NPM mutations in a retrospective cohort of 106 patients with normal karyotype treated according to the ALFA-9000 or ALFA-9802 protocols between 1990 and 2004. In these patients aged 17–65 years, NPMm was significantly associated with a high white blood cell count (69 vs 18 G/L, p&lt;.001) and an involvement of the monocytic lineage (FAB AML-M4/M5, p=.009). NPMm was also associated with a low rate of CEBPA mutation (CEBPAm, 10% vs 27%, p=.05) and a higher rate of FLT3 internal tandem duplication (FLT3/ITD, 38% vs 25%, p=NS). We did not find any correlation between NMPm and CR rate (86% vs 88%, p=NS). Long-term outcome did not differ between NPMm and NPM wild-type (NPMwt) patients neither in univariate analysis (6y-OS, 43% vs 37%; 6y-RFS, 47% vs 34%, p=NS) nor in multivariate analysis after adjustment on covariates significantly associated with prognosis in univariate analysis (age, CEBPAm, FLT3/ITD). NPM mutational status was available for 15 patients at relapse time. Ten patients out of 15 had NPM mutations at diagnosis and still displayed NPM mutations at relapse time. None of the 5 NPMwt patients acquired NPM mutation at relapse. Recently, we developed a RQ PCR technique to study minimal residual disease. Fifteen patients were evaluated at diagnosis and follow-up. In the majority of the cases a sensitivity of 10−4 was obtained. The high frequency, the stability, and the homogeneity of NPMm provide a promising minimal residual disease marker that we are prospectively evaluating. Prospective studies are also needed to confirm the definitive role of NPM mutation in the prognosis of AML patients.
FIP1L1-PDGFRA Positive Secondary Acute Myeloid Leukemia.
Authors: Metzgeroth Georgia , Popp Helena , Walz Christoph , Müller Martin C. , Mix Jürgen , Paschka Peter , Müller Lothar , Steder Martin , Del Valle Francisco , Aulitzky Walter E. , Benke Harro , Schoch Claudia , Haferlach Torsten , Hehlmann Rüdiger , Hochhaus Andreas , Reiter Andreas 
Published: April 2020

Abstract
The FIP1L1-PDGFRA positive myeloproliferative syndrome (MPS) represents a distinct clinicobiological entity with constitutively enhanced tyrosine kinase activity of the fusion protein and excellent response to treatment with imatinib. Most patients are initially diagnosed as idiopathic hypereosinophilic syndrome or chronic eosinophilic leukemia (CEL). A prominent clinical feature besides eosinophilia is the male predominance. However, the natural clinical course remains to be elucidated. Here we report on five male FIP1L1-PDGFRA positive patients (median age 58, range 46–64 years) with various subtypes of acute myeloid leukemia (AML) according to the FAB classification (M0, n=2; M2, n=1; M4eo, n=1; acute eosinophilic leukemia, n=1). All patients had a history of pronounced eosinophilia for a median interval of 6 mo (range 2–186) indicating the presence of a CML-like chronic phase disease evolving to blast crisis or secondary AML. One patient relapsed six years after conventional chemotherapy of a secondary FIP1L1-PDGFRA positive eosinophilia-associated AML M0 with typical features of CEL. Three patients had additional cytogenetic aberrations at diagnosis of AML similar to those frequently observed during clonal evolution of CML (trisomy 8, n=2; trisomy 9, n=1). All five patients received imatinib (100–300 mg/day) for a median time of 12 mo (range 1–23); three patients as monotherapy, one patient after one course and another patient after two courses of intensive chemotherapy. Three AML patients have been treated for more than 3 mo (12+, 18+, and 23+ mo) and are currently free of relapse. All three patients achieved a rapid complete hematological remission, defined as a complete clearance of blasts, normalization of peripheral blood and bone marrow eosinophil count, and a complete molecular remission 4, 5, and 14 mo, respectively, after start of treatment as determined by nested RT-PCR for the FIP1L1-PDGFRA transcript. In all cases treatment with imatinib was well tolerated. We conclude that the occurrence of the FIP1L1-PDGFRA fusion gene seems to be associated with a chronic phase disease which may progress to secondary AML. Therefore, all patients presenting with an eosinophilia-associated AML and normal karyotype should be screened for the cytogenetically invisible FIP1L1-PDGFRA fusion gene by RT-PCR and/or fluorescence in situ hybridization, in addition to the commoner CBFB-MYH11 fusion. Patients with a FIP1L1-PDGFRA positive MPS are excellent candidates for treatment with imatinib even if they present with secondary AML.
Prognosis Value of Flt3 Internal Tandem Duplication and P-Glycoprotein Functionality, in 171 AML.
Authors: Marzac Christophe , Teyssandier Irene , Perrot Jean Yves , Faussat Anne Marie , Tang Ruoping , Casadevall Nicole , Marie Jean Pierre , Legrand Ollivier 
Published: April 2020

Abstract
Adult acute myeloid leukemia (AML) with intermediate cytogenetic remains a very heterogeneous group of patients with highly variable individual prognosis. Emerging data suggest that some molecular markers could help to refine cytogenetic stratification. Here we focused on ABCB1 (MDR1/Pgp) ATP-binding cassette transporter activity and fms-like tyrosine kinase mutations (Flt3/ITD) because their individual prognosis value is well demonstrated and because they may lead to targeted therapy. Therefore we assessed Pgp activity using JC-1 probe and Flt3 exon 14 mutational status by standard PCR and realised a multivariate analysis in 171 adult AML patients treated in the EORTC protocols. Flt3/ITD (ITD+) and high Pgp activity (Pgp+) were found in 26/171 (15%) and 55/171 (32%) of the patients, respectively. The repartition was remarkable in that, as defined with our criteria, both were negative in 94/171 (55%, Pgp-ITD- group) or mutually exclusive in 73/171 (43%, Pgp-ITD+ and Pgp+ITD- groups) and only 4/171 (2%, Pgp+ITD+ group) were positive for both. In univariate analysis complete remission (CR) rates for ITD+ were 38% vs 61% for ITD- patients (p=0.034) and 47% vs 62% (p=0.06) for Pgp+ vs Pgp- patients. Individually, both parameters were strong predicators for overall survival (OS) (p=0,02) but not for disease free survival. In multivariate analysis Pgp+ITD+ (p&lt;0.0001) and age (p=0.0022) were independent prognostic factors for CR achievement. For OS, CR achievement (p&lt;0.0001), WHO performance status (p=0.0007) and Pgp+ITD+ (p=0.0014) were also independent prognostic factors. In 98 patients with intermediate cytogenetic CR rates for ITD+ were 40% vs 62% for ITD- patients (p=0.099) and 41% vs 67% (p=0.014) for Pgp+ vs Pgp- patients. In the Pgp-ITD- group (47/98 patients), CR rate was 70% vs 44% for others (p= 0.012). The 4 years OS achieved 48% vs 16% (p&lt;0.0001) and DFS was 56% versus 27% (p=0.024). Furthermore, OS curves of the intermediate cytogenetic-Pgp-ITD- group were not significantly different from the favorable cytogenetic group. In conclusion Flt3-ITD and P-gp activity are independent and additive prognostic factors, which provide a powerful risk classification that can be used to stratify the treatment of intermediate cytogenetic AML patients.
Figure Figure
Exon-12 Nucleophosmin (NPM) Mutation and Aberrant Cytoplasmic Expression of NPM Protein in Leukemia Cell Line OCI-AML3.
Authors: Quentmeier Hilmar , Martelli Maria P. , Dirks Wilhelm G. , Bolli Niccolo , Liso Arcangelo , MacLeod Roderick A.F. , Nicoletti Ildo , Mannucci Roberta , Pucciarini Alessandra , Bigerna Barbara , Martelli Massimo F. , Mecucci Cristina , Drexler Hans G. , Falini Brunangelo 
Published: April 2020

Abstract
Wild-type nucleophosmin (NPM) is a multifunctional protein shuttling between the nucleus and the cytoplasm. Chromosomal rearrangements leading to NPM fusion proteins occur in leukemias and lymphomas (e.g. with partners RARA, ALK). Recently, Falini et al. reported that 60% of acute myeloid leukemia (AML) patients with normal karyotype carry mutations at exon-12 of the NPM gene. This results in frame shifts that lead to alterations of the C-terminus of NPM resulting in the aberrant cytoplasmic localization of the mutated protein (NPMc+) (1). The effects of a mutationally altered protein on cellular functions like proliferation, differentiation or apoptosis, have often been revealed using immortalized cell lines that carry the mutation in question. Therefore, we screened a panel of 79 myeloid leukemia cell lines for presence of mutations - 4 bp insertions - at the exon-12 of the NPM gene. We performed polymerase chain reaction (PCR) analysis with fluorescent dye-labeled primers. For fragment size determination, the PCR products were mixed with dye-labeled size standards and separated by capillary electrophoresis. OCI-AML3 was the only cell line that expressed a signal in addition to and 4 bp larger than the wild-type NPM signal. Sequencing of the cloned NPM-mutated PCR product showed TCTG duplication at positions 956–959 of exon-12. This mutation was heterozygous and corresponded to the type that occurs in 77% of primary NPMc+ AMLs. OCI-AML3 cells have a hyperdiploid karyotype with 48(45–50)&lt;2n&gt;X/XY, +1, +5, +8, der(1)t(1;18)(p11;q11), i(5p),del(13)(q13q21), dup(17)(q21q25); sideline with r(Y)x1-2 and show the following immunoprofile: CD3−, CD4+, CD7−, CD8−, CD10−, CD13+, CD14−, CD15+, CD19−, CD30−, CD33−, CD34−, CD41+, CD42b−, CD68+, CD235a+, HLA-DR-. Especially the presence of myeloid markers and absence of CD34 is typical for NPMc+ cells (1). Furthermore, immunostaining with anti-NPM antibodies confirmed that the OCI-AML3 cells, like primary NPMc+ AML and in contrast to NPM wild-type cells, show cytoplasmic expression of NPM. Functional studies showed that the altered nucleo-cytoplasmic transport of NPM was nuclear export signalling-dependent and could be blocked by using the specific CRM1/exportin-1 inhibitor leptomycin B. In conclusion, cell line OCI-AML3 promises to be an important tool for studying the biological properties and response to new drugs of NPMc+ AML.
IL-7 Differentially Regulates Cell Cycle Entry and Exit of Naive and Memory CD4+ T Lymphocytes: Effects on HIV-Mediated Infection.
Authors: Verhoeyen E.L.S. , Swainson Louise , Cosset Francois-Loic , Taylor Naomi 
Published: April 2020

Abstract
The IL-7 cytokine plays a major role in the in vivo maintenance of polyclonal naive and memory T-cells, positively regulating the survival, differentiation and proliferation of thymocyte and peripheral T lymphocyte populations. Under conditions of lymphopenia due to HIV infection, it has been shown that increased peripheral IL-7 levels correlate with T cell differentiation. Moreover, a short-term exogenous IL-7 treatment enhances T-cell differentiation as well as expansion of naive and memory T-cells. These properties have lead to the proposal that recombinant IL-7 be used as an adjuvant immune therapy in HIV-infected individuals. Nevertheless, primate studies have shown that after an initial increase in in vivo T cell proliferation, there is a substantial drop in the absolute lymphocyte numbers (Fry et al., 2003; Nugeyre et al., 2003). As the bases for this decrease have not yet been elucidated, it is important to assess the long-term biological effects of IL-7 on quiescent human CD4+ T lymphocyte subsets. Here, we demonstrated that although IL-7-stimulated memory T-cells enter into cycle much more rapidly than their naive counterpart (3 days as compared to 6 days), they also exit the cell cycle much earlier, by day 10 vs. day 18. Importantly, cell cycle exit occurred despite the continuous replenishment of IL-7 and was inversely correlated with IL-7 receptor levels. Specifically, IL-7Rα levels were completely downregulated on both naive and memory T subsets within 12 hours post cytokine treatment but were significantly re-upregulated in memory T cell within 8–10 days. In naive lymphocytes, IL7Rα expression was absent during the first 14 days of continuous IL-7 stimulation after which time it slowly increased, with expression coinciding with cell cycle exit. Importantly, the permissivity of IL-7-stimulated CD4+ lymphocytes to infection with an HIV-1 vector was not related to cell cycle entry per se. After extended IL-7 stimulation of the naive T cell subset, they remained more refractory to HIV-1 vector infection than memory cells even though they demonstrated a higher level of cell cycle progression. Moreover, under conditions mimicking the lymph node environment, cell cycle entry was not required for IL-7 mediated infection. The differential effects of recombinant IL-7 on the cell cycle entry-exit status of naive and memory CD4+ T lymphocytes as well as the relative susceptibilities of these CD4+ subsets to HIV-1 vector infection have important implications for the use of this cytokine as an adjuvant immunotherapy.
Adoptive T Cell Therapy Using Antigen-Specific CD8+ T Cells for the Treatment of Patients with Cancer: A Phase I Clinical Study.
Authors: Mackensen Andreas , Meidenbauer Norbert , Vogl Sandra , Laumer Monika , Andreesen Reinhard 
Published: April 2020

Abstract
The adoptive transfer of in vitro induced and expanded tumor antigen-specific cytotoxic T lymphocytes (CTL) provides a promising approach to the immunotherpy of cancer. We have previously shown that antigen-specific CTL can be generated from HLA-A2.1+ cancer patients by 4 rounds of in vitro stimulation of purified CD8+ T cells with autologous dendritic cells pulsed with HLA-A2 binding tumor-associated peptides. Based on these results we have initiated a pilot study of adoptive T cell therapy in advanced melanoma patients demonstrating that in vitro generated Melan-A (ELAGIGILTV)-specific CTL survive intact in vivo for several weeks and localize preferentially to tumor. Here we report on the clinical results of a phase I study of 11 HLA-A2+ melanoma patients that received at least three i.v. infusions of Melan-A-specific CTL i.v. at 2-week intervals. Each T cell infusion was accompanied by a 6-day course of s.c. IL-2 (3x106 IU daily). A total of 51 T-cell infusions were administered, averaging 2.1 x108 Melan-A specific T cells per infusion, with a range from 0.11 – 13.1 x108 T cells per infusion. Clinical side effects were mild and consisted of chills and low-grade fever (WHO grade I–II) in 7 out of 11 patients that typically occurred within 6 to 8 h post infusion. Hematological effects, observed after T cell transfer, included an increase in eosinophils up to 50% in 7 out of 11 patients, peaking 24h post transfer. Clinical and immunological responses consisted of antitumor responses in 3 out of 11 patients (1 CR, 1 PR, 1 mixed response), an elevated frequency of circulating Melan-A multimer+ T cells up to 2% of total CD8+ T cells for 2 weeks post T cell infusion, suggesting long-term survival and/or proliferation of transferred CTL, and a complete loss of Melan-A expression in lymph node metastases of 2 patients after T cell transfer. Our data indicate that the adoptive transfer of antigen-specific T cells in cancer patients is capable of inducing clinical and systemic tumor-specific immune responses without provoking major side effects.