bluuebunny/crossref_metadata_2025_split huggingface dataset:
Raf1 Is Required for Induction of a Bcr-Abl Positive CML Like Myeloproliferative Disease in Mice
Authors: Albers Corinna , Illert Anna Lena , Miething Cornelius , Peschel Christian , Duyster Justus 
Published: April 2020

Abstract
Introduction: Chronic myelogenous leukemia (CML) results from neoplastic transformation of hematopoietic stem cells (HSC), characterized by a chromosomal translocation t(9;22)(q34;q11). This aberration leads to the expression of the oncogenic tyrosine kinase Bcr-Abl, which mediates signals for proliferation, transformation and anti-apoptosis via various different pathways including the Raf/MEK/ERK cascade. The cytoplasmic protein Raf1 is a key molecule within this cascade. Recent studies have revealed an additional function of the Raf-1 kinase that is independent of the activation of the MAPK cascade and whose effect is to increase resistance to apoptosis. Therefore Raf1 is an interesting target for molecular therapies and more effective Raf1 inhibitors have recently been developed by the pharmaceutical industry. Here we report the impact of Raf1 signalling for Bcr-Abl mediated transformation.
Methods: We exerted a siRNA based approach in combination with a murine bone marrow transplantation model. To this end we designed a MSCV based retrovirus encoding both the Raf1 microRNA and the Bcr-Abl oncogene on a single construct. This approach ensured knockdowns of more than 90% of Raf1 in every Bcr-Abl transformed cell.
Results: Methylcellulose assays demonstrated that bone marrow coexpressing Raf1 microRNA and Bcr-Abl had a 2 fold decreased colony forming ability compared to control cells. We then transduced bone marrow (BM) with retrovirus coexpressing Raf1 microRNA and p185 Bcr-Abl and transplanted lethally irradiated recipient Balb/C mice. The onset and progression of leukemia was significantly delayed in mice transplanted with Raf1 microRNA and Bcr-Abl compared with the Bcr- Abl transduced control microRNA group. Raf1 knockdown mice showed only a moderate rise of white blood cell (WBC) counts and prolonged overall survival (median survival 39 ± 7.1 days) in comparison to control mice (23.3 ± 2.4 days). However, we were not able to completely avoid the development of leukemia by Raf1 knockdown.
Conclusion: Taken together our data demonstrate that Raf1 is important for the development of a myeloproliferative disease by Bcr-Abl in mice. Therefore Raf1 inhibition in combination with Bcr-Abl kinase inhibition depicts an interesting approach towards eradication of Bcr- Abl positive leukemia. In addition, this study describes a novel and versatile approach to express an oncogene and a microRNA using a single retroviral construct. Thus this powerful tool can be used to systematically screen drugable signalling targets involved in oncogenesis.
Nilotinib in Elderly Chronic Myeloid Leukemia Patients in Chronic Phase (CML-CP) with Imatinib Resistance or Intolerance: Efficacy and Safety Analysis
Authors: Lipton Jeffrey H. , le Coutre Philipp D. , Wang Jim , Yang Mindy , Szczudlo Tomasz , Giles Francis 
Published: April 2020

Abstract
Background: Nilotinib is a potent and highly selective BCR-ABL kinase inhibitor approved for the treatment of Philadelphia chromosome-positive chronic myeloid leukemia (Ph+ CML) patients in chronic (CML-CP) or accelerated phase (CML-AP) who have failed prior therapy including imatinib.
Methods: This subanalysis of the open-label, single-arm, phase 2 study evaluated the efficacy and safety of nilotinib in elderly (≥65 years) CML-CP patients who were resistant or intolerant to imatinib. Nilotinib was dosed at 400 mg twice daily.
Results: A total of 321 CML-CP patients (71% imatinib-resistant; 29% imatinib-intolerant) were enrolled. Thirty percent (98/321) of patients were ≥65 years; 8% (8/98) of these patients were 380 years. The baseline characteristics among patients ≥65 and &lt;65 years were similar. Discontinuation of nilotinib due to adverse events was uncommon, and did not differ among the two age groups (18% in both groups). Efficacy was maintained in elderly patients, with 48% of patients achieving MCyR and 38% achieving CCyR, compared with 63% and 44% of patients &lt;65 years achieving MCyR and CCyR, respectively. Duration of cytogenetic response was also consistent among age groups with 84% and 85% of responding elderly patients maintaining MCyR and CCyR at 18 months, compared with 85% and 89% of patients &lt;65 years maintaining MCyR and CCyR at 18 months, respectively. Estimated overall survival (OS) rates at 12 months were 97% and 91%, for patients &lt;65 years and ≥65 years, respectively. Overall, the safety profile of nilotinib was similar among the two age groups. Biochemical laboratory abnormalities were transient, clinically asymptomatic, and consistent in both age groups; elevated lipase occurred in 14% and 23%, and elevated total bilirubin occurred in 9% and 3% of patients &lt;65 years and ≥65 years, respectively. The most common grade 3/4 hematological laboratory abnormalities were also comparable; neutropenia was reported in 31% and 30%, thrombocytopenia in 26% and 36%, and anemia in 8% and 15%, of patients &lt;65 years or ≥65 years, respectively. The incidence of grade 3/4 pleural/pericardial effusions were &lt;1% and 1% and grade 3/4 bleeding events were &lt;1% and 1% in patients &lt;65 years or ≥65 years, respectively. The incidence of myocardial infarction (&lt;1% vs 4%), congestive heart failure (2% vs 1%), and QTcF prolongation &gt;500 msec (&lt;1% vs 2%) were also similar among patients &lt;65 years or ≥65 years.
Conclusions: Nilotinib is highly active and induced durable clinical responses in CML-CP patients regardless of age. Importantly, the safety profile of nilotinib is maintained in elderly patients and there was no increase in the incidence of cardiac evens making it an excellent therapeutic option for patients with Ph+ CML, regardless of age.
The Incidence and Outcome of Acute Lymphoblastic Leukemia in Shanghai, China Over 5 Years (2002–2006)
Authors: Ni Xiong , Shen Zhixiang , Chen Fangyuan , Liang Hui , Lu Fengjuan , Tang Jingyan , Wang Chun , Shao Jingbo , Hou Jian , Zou Shanhua , Wang Jianmin 
Published: April 2020

Abstract
Great advances have been made in the diagnosis, molecular pathogenesis and treatment for acute lymphoblastic leukemia (ALL) in the past decade. Due to the lack of comprehensive population-based studies, the updated trend of incidence and geographic variations of ALL has not been well documented in Shanghai, China. To better understand the incidence and epidemiological features of ALL in Shanghai, we conducted a survey based on the database in the Center for Disease Control and the medical record in all hospitals in Shanghai, especially 28 major hospitals with hematology department. According to the CDC data, 544 patients with median age of 32y (1.2~89y) were diagnosed as de novo ALL during January 1, 2002 and December 31, 2006 and followed-up until December 31, 2007. The yearly incidence of ALL was (0.81±0.11)/100 000, which is slightly higher in men (0.86/100 000) than in women (0.75/100 000). The age-stratified incidence showed 2.31/100 000 in pts ≤17y, 0.54/100 000 in 18~34y, 0.46/100 000 in 35~59y, and 0.94/100 000 in pts &gt; 60y. There were substantial geographic variations in the incidence of ALL, with 0.57±0.14/100 000 in Chongming county, an island eastern of Shanghai city, much lower than that of other districts in the central or around the central city, such as Qingpu, Baoshan, Hongkou, and Jiading districts (&gt;0.9/100 000, P&lt;0.05). Both FAB and WHO classification system were applied in the present study. Eighty-eight pts were diagnosed as L1 (26.2%), 193 L2 (57.4%), and 55 L3 (16.4%), For 302 pts with immunophenotyping data, 242 pts were identified as B cell orgin (80.1%), 59 T cell origin (19.5%), and 1 biphenotype (0.4%). The leukemia cells in 61 pts co-expressed one or two myeloid antigen (20.2%). For 269 pts with cytogenetics data, the incidence of t (9;22) in pts aged &lt;10y, 11–17y, 18–44y, 45–59y and ≥60y were 4.2%, 11.4%, 19.2%,23.1% and 5.3%, respectively. For 473 evaluable pts, the complete remission (CR) rate and 3 year OS was 72.4% and (34.2±2.3)%, respectively. Both CR rates and 3 year OS decreased with age, 88.8% and (75.9±4.1%) in pts aged≤10, 87.3% and (36.8±6.8)% in 11–17y, 78.0% and (27.2±4.5)% in 18–44y, 73.5% and (16.1±4.7)% in 45–59y and 35.1% and (2.7±1.8)% in ≥60y (P&lt;0.01). In this cohort of pts, only 26 received allogeneic hematopoietic stem cell transplantation (Allo-HSCT) in CR1. The probability of 3-year OS was 59.9±9.4%, better than those treated with conventional chemotherapy (36.3±8.0%) or auto-HSCT (33.3±17.2%, P&lt;0.05). The age, WBC count at diagnosis, cytogenetic changes and donor availability are the factors influencing the prognosis.
Clofarabine in Combination with Etoposide and Cyclophosphamide (CLOVE) in Pediatric Patients with Relapsed Acute Leukemia: 4 Italian Pediatric AIEOP Centers’ Experience
Authors: Micalizzi Concetta , Parasole Rosanna , Putti Maria Caterina , Luciani Matteo , Banov Laura , Petruzziello Fara , Messina Chiara , De Rossi Giulio 
Published: April 2020

Abstract
Clofarabine is a new promising chemotherapic agent active in the treatment of pediatric acute leukemia. We planned to utilize the same drugs’ combination currently evaluated in USA as phase II study salvage therapy: clofarabine (40mg/m2/day) in combination with etoposide (100mg/m2/day) and Cyclophosphamide (440mg/m2/day) (CLOVE). We planned to treat the patients (pts) (aged 1–18) with relapsed or refractory acute lymphoblastic leukemia (ALL) or acute myeloblastic leukemia (AML) with 1 or 2 induction cycles (5 days chemotherapy) to reach complete remission (CR) or complete remission without platelet recovery (CRp) and with 1 or 2 consolidation cycles (4 days chemotherapy). ALL patients (pts) received intrathecal methotrexate at day 6 and AML patients intrathecal cytarabine at Day 0. Prednisone was added (0,5 mg/Kg/day) to prevent systemic inflammatory response syndrome (SIRS). Fourteen children (5 AML, 9 ALL) were treated, median age 5,7 years (range 1,75–16,5), 7 pts in first, 3 pts in second and 4 pts in third relapse. Four AML pts had one prior bone marrow transplantation (BMT): 1 autologous, 1 matched familiar donor and 2 matched unrelated donor (MUD) transplants. Five ALL pts had one prior BMT : 2 autologous, 1 syngenic and 2 MUD. The AML pts were in first relapse, and three of them were refractory to FLAG-X protocol: 3/5 (60%) achieved remission (1CRp, 2 CR), response data are currently pending for1 patient (day + 20 first induction), and 1 patient died for disease progression. The remission rate of nine ALL pts was 66% (4 CR, 2CRp). Infection grade 3 was observed in one patient, diarrhea and abdominal pain occurred in 4 pts. Febrile neutropenia was e common toxicity. Severe muscles pain with functional impairment was observed in two pts (1 AML, 1 ALL) 7 days post induction cycle: the gabapentin therapy in association with prednisone was efficient and the symptom quickly disappeared. One ALL patient refractory to first induction cycle, died after second cycle (day + 6) for cardiac failure. One ALL patient relapsed after CLOVE induction, after two more cycles he obtained partial response (BM Blast cells 10% ), and he underwent MUD transplantation and died for infection. The overall survival is 58 %, 7/9 (77 %) responders are alive. The median survival is 3 months (range 20 days –10 months). Four out 9 responding patients proceeded to BMT from unrelated donor, and three of them are alive and in CR. No unexpected transplant related toxicity was encountered. We confirm that the association of Clofarabine with Etoposide and Cyclophosfamide is active in heavily pretreated relapsed or refractory pediatric acute leukemia. patients.
Long-Term Monitoring of Patients with Hematological Malignancies and Hepatitis B and Hepatitis C Virus Infections
Authors: Garmaeva Tatyana , Kulikov Sergei , Michailova Elena , Sudarikov Andrey , Filatov Felix , Savchenko Valeryi 
Published: April 2020

Abstract
INTRODUCTION. Patients (pts) with hematological malignancies are at high risk of infection by Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) due to multiple transfusions and the large number of invasive procedures. The aim of the study was to evaluate the presentation rate of clinical hepatitis, pt survival and how hepatitis infection influenced survival.
RESULTS. The study group was comprised of all pts admitted to the hematology department from Feb 2004 to June 2006. The pts were followed until June 2008. All pts were monitored by testing of HBsAg, anti-HCV, DNA-HBV, RNA-HCV, HBeAg, anti-HBs, anti-HBc, anti-HBe approximately every 3 weeks. Liver biopsy was performed on 64 pts, 23 pts each with HBV and HCV immunohistology. A total of 7800 biological samples were collected for HBV and HCV testing; of these, 4000 were tested by PCR methods. Acute leukemias (AL) and aplastic anemias (AA) constituted 77% (205/265) of all pts. The median age was 38 years (range, 15 to 79), Male; 47% (n=125), female; 53% (n=140). Median transfusion load (the number of donors per pt for the study period) was 45 (range, 0 to 418). When patients new to the hematology dept were examined: HBV was detected in 15% (39/265) of new pts and HCV in 7% (19/265) of new pts respectively. We postulated that the rate of infection is extremely high in the hematology clinic. For the entire study period from Feb 2004 to Jun 2006, 51% (135/265) pts had positive markers for HBV infection, 19% (51/265) pts had positive markers for HCV, and 14% (37/265) pts had positive markers for both HBV+HCV. We have shown that up to 95% of HCV-positive pts and up to 60% of HBV-positive pts developed clinical and biochemical symptoms and signs of viral hepatitis with 3 years of initial detection of HBV and HCV markers. 154 (58%) pts survived, 111 (42%) pts died. Of these, 2 pts developed fulminant liver failure due to severe hepatitis B. The analysis of survival risk factors demonstrates that the expected life duration significantly decreases after HBV infection. For AL pts RR=1.8 (p=0,034), for AA pts RR=4.3 (p=0,022). There was no significant association between the expected life duration and HCV infection. Proportional hazard regression model with time dependent covariates (PHREG SAS) was used for the analysis.
CONCLUSION. The majority of pts infected by HBV and HCV developed clinically recognizable viral hepatitis within 3 years from the first detection of viral markers. Pts with severe immunosupression often do not manifest obvious features of acute viral hepatitis. Pts with hematological malignances should be monitored regularly (at least once in a month) for HBV and HCV markers during all period of treatment and for 1–2 years after completion of therapy. Viral hepatitis B is a risk factor associated with survival risk factor for AL and AA pts.
An Unusual Presentation of Acute Biphenotypic Leukemia without Bone Marrow Involvement
Authors: Bayraktar Ulas D , Escalon Maricer 
Published: April 2020

Abstract
Acute biphenotypic leukemia (ABL) represents a minority of acute leukemia cases (around 5%) in which blasts co-express markers of different lineages (myeloid, T-cell or B-cell). ABL without bone marrow involvement is exceedingly rare. Case report: An 18-year-old man presented with slowly enlarging, non-tender cervical lymph nodes. He had no other symptoms. Family history included Hodgkin’s disease in his mother and maternal great aunt, histiocytosis X in his maternal cousin, and non-Hodgkin’s lymphoma in his paternal grandfather. Physical exam revealed multiple small cervical and axillary lymph nodes. Laboratory analyses including complete blood count, peripheral blood smear, complete metabolic profile, mononucleosis screen, and hepatitis and HIV serologies were unremarkable. Histopathological examination of the left cervical lymph node revealed rare germinal centers with an interfollicular proliferation of generally medium size cells with a fine blastic chromatin pattern and scant cytoplasm. CD34 was positive in more than half of the cells. Between 30% and 80% of the cells were positive for TdT (Figure 1), CD3, CD4, and CD7. Myeloperoxidase (Figure 2) and lysozyme were positive in about 20% of the cells. Flow cytometry revealed an immature myeloid cell population coexpressing CD33, CD13, CD7, and CD34. Chromosomal analysis revealed a complex karyotype. Bone marrow exam was unremarkable and no evidence of leukemia was observed. Flow cytometry of the bone marrow aspirate revealed only 4% of the cells to be CD34+. Whole body PET scan showed extensive uptake in bilateral axillae, neck, and inguinal regions without any extranodal involvement. The patient received hyper-CVAD regimen (alternating cycles of cyclophosphamide, vincristine, adriamycin, dexamethasone and high dose methotrexate, cytarabine) for 8 cycles and remains in remission five months after the end of hyper-CVAD protocol. To our knowledge, this is the third case reported in the literature with acute biphenotypic leukemia not involving the bone marrow. However, our case cannot accurately be referred to as leukemia since the bone marrow is not involved; neither can it be named as lymphoblastic lymphoma because of its myeloid differentiation. Therefore, we will refer to it as extramedullary granulocytic sarcoma with biphenotypic features.
Figure 1 Figure 1. Figure 2 Figure 2.
Inhibition of Long-Term Expansion in a Subgroup of Acute Myeloid Leukemia Samples by Dasatinib
Authors: Han Lina , Schuringa Jan Jacob , Mulder Andre , Vellenga Edo 
Published: April 2020

Abstract
The Src family kinases are frequently constitutively activated in acute myeloid leukemia (AML) due to mutations in receptor tyrosine kinases including Flt3 and c-Kit or to the autocrine or paracrine production of growth factors. Recent studies have demonstrated that dasatinib is an effective inhibitor of Src kinase activity in chronic myeloid leukemia (CML) by blocking the Bcr/Abl-mediated Src activity resulting in inhibition of cellular proliferation. In the present study we questioned whether similar effects can be observed in AML cells exposed to dasatinib and whether differences can be observed in normal cord blood (CB) CD34+ cells and CML cells (K562 cell line). Firstly, we demonstrated that dasatinib was effective in blocking the proliferation of K562 cells at a concentration of 2–10 nM which coincided with a significant reduction in Src, Erk1/2 and Stat5 phosphorylation. Long-term MS5 cocultures initiated with CB CD34+ cells demonstrated a reduced expansion of 50% at 5 nM dasatinib (p=0.005, n=3) after 4 weeks of coculture which was associated with a reduction in progenitor output as determined by colony forming cell (CFC) assays (p=0.031). Moreover, the inhibitory effects of dasatinib were cytokine-specific, eg IL3 and TPO-induced proliferation of both CB and Mo7e cells were not inhibited by dasatinib, while SCF-mediated proliferation was significantly impaired (94% ± 4% inhibition, p&lt;0.001) at 5 nM. These inhibitory effects were associated with a reduced SCF-induced phosphorylation of Erk1/2 and Stat5 in the presence of dasatinib. Subsequently, AML cells (n=19) were studied in the long-term stromal culture assays by using exclusively the sorted CD34+ cell fraction that is enriched for leukemic stem cells. In 79% (15/19) of the tested AML cases long-term expanding AML co-cultures could be generated. In 27% (4/15) of the AML cases a distinct reduction in cell proliferation of AML-CD34+ cells was observed as compared with the untreated group, ranging from 48%–91% inhibition at low concentration of dasatinib (0.5 nM). This concentration of dasatinib did not significantly affect expansion of normal progenitors in the long-term culture assays (36% ± 12% inhibition, p=0.15). The AMLs that responded to dasatinib were characterized by Bcr-Abl (n=1), c-Kit (n=1), Flt3-ITD (n=1) or undetermined (n=1) mutations. The additional Flt3-ITD+ AMLs (n=6) did not show a reduced expansion to dasatinib exposure. No difference in Src expression was observed between the responsive and non-responsive AMLs. In conclusion, our results demonstrate that dasatinib is effective in blocking SCF-mediating signaling in hematopoietic cells which is associated with reduced phosphorylation of Erk1/2 and Stat5, while in the malignant counterpart long-term expansion of a subgroup of AML CD34+ cells can be inhibited at low-doses.
Influence of Genetic Polymorphisms of VKORC1 and CYP2C9 in Patients on Phenprocoumon Steady-State Dose Requirements
Authors: Wu Weidong , Harenberg Job , Weiss Christel , Kirchheiner Julia , Stehle Simone , Fuhr Uwe , Gleiter Christoph 
Published: April 2020

Abstract
The variants of the (Vitamin-K epoxid reductase subcomplex 1) VKORC1 and of cytochrome P450 2C9 (CYP2C9) genes are know to influence the steady state dose of the vitamin-K antagonist (VKA) phenprocoumon during oral anticoagulant therapy. We have analysed in detail the influence of the VCORC1 1173CC and 1173CT variants in combination with the CYP2C9 variants on the steady state dose of phenprocoumon. The weekly phenprocoumon dosage was calculated after a 3 to 6 months of stable anticoagulation by the international normalized ratio (INR) which had to be in a range from 2 to 3 in 114 patients. VKORC1 1173CC, CT, TT and CYP2C9*1/*1, *1/*2, *1/*3, *2/*2, *2/*3 and *3/*3 genotypes were determined using direct DNA sequencing and realtime PCR. Patients carrying VKORC1 1173CT and CYP2C9*1/*1 genotypes required a 33% lower steady state dose of phenprocoumon compared to VKORC1 1173CC and CYP2C9*1/*1 genotypes (p=0.0001). Patients with VKORC1 1173CC in combination with CYP2C9*1/*2, *1/*3 needed 10% and 34% lower steady state dose of phenprocoumon compared to the combination with CYP2C9*1/*1, respectively (p=0.0001). Patients carrying VKORC1 1173CT needed lower doses of phenprocoumon in combination with CYP2C9*1/*1 (−32%), *1/*2 (−29%), or *1/*3 (−33%), and VKORC1 1173TT in combination with CYP2C9*1/*3 (−44%) compared to VKORC1 1173CC combined with CYP2C9*1/*1 (all p&lt;0.001). CYP2C9 variants did not influence the dose of carriers of VKORC1 1173CT gene. Both genotypes together influenced the phenprocoumon maintenance dosage in this naturalistic study design. Genotyping for CYP2C9 and VCORC1 therefore might be a suitable tool for dose individualization in patients at oral anticoagulant drug therapy.