bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Analysis of FLT3 and NPM1 Gene Mutations in Chinese Patients with Acute Myeloid Leukemia.
Authors: Zhang Su-Jiang , Li Jianyong , Han Yangli 
Published: October 2019

Abstract
Abstract 4136

Objective
To investigate the frequency of nucleophosmin(NPM1) gene mutations and FLT3 mutations in Chinese patients with acute myeloid leukemia (AML) and its correlation with clinical feature and prognosis.

Methods
123 first diagnosed AML patients were involved in our study. Polymerase chain reaction (PCR) combined with electrophoresis was directly used to detect FLT3-ITD mutations, PCR combined with EcoRV digestion was used to detect FLT3-TKD mutation, and PCR combined with directly sequencing was used to detect NPM1 mutations.

Results
(1) In the 123 first diagnosed AML patients, 16 patients (13.0%) were found harboring FLT3-ITD mutation, 6 patients (4.9%) were FLT3-TKD positive, and one patient was found harboring co-existing FLT3-ITD and FLT3-TKD mutation. In addition, 24 patients (19.8%) were NPM1 positive, including 22 patients with A-type mutation, one patient with B-type mutation and one patient with D-type mutation. 5 patients were found harboring co-existing FLT3-ITD and NPM1 mutation. (2) FLT3-ITD and NPM1 mutations were identified as 17.5% (10/57) and 31.6% (18/57) individually in 57 AML patients with normal karyotype, which were significantly higher than patients with abnormal karyotype. All of patients with double mutation had normal karyotype. (3) White blood cell (WBC) count and leukemia cell of bone marrow in FLT3-ITD or FLT3-ITD+/NPM1+ patients were both higher than those patients with FLT3-ITD-/NPM1-. The CR rate of FLT3-ITD+/NPM1-, FLT3-ITD+/NPM1+, FLT3-ITD-/NPM1+, FLT3-ITD-/NPM1- were 50% (4/8), 20% (1/5), 87.5% (7/8), 79.5% (35/44) individually.

Conclusions
FLT3 gene and NPM1 gene mutations are common in AML patients, and early detection of FLT3 gene and NPM1 gene mutations will be beneficial for treatment and prognosis analysis of AML patients.

Disclosures:
No relevant conflicts of interest to declare.
HL-60 Cell Death Induced by Lycorine Is Due to p21-Mediated Cell Cycle Arrest and TNF-α Signaling Stimulation.
Authors: Liu Jing , Hu Ji-liang , He Yang , Shi Bi-Wei , Hu Wei-Xin 
Published: October 2019

Abstract
Abstract 4158
Lycorine displays various biological functions including remarkable anti-tumor effect. We previously reported that lycorine induced anti-leukemia effect via arresting cell cycle and inducing apoptosis on human acute promyelocytic leukemia (APL) cell line HL-60. To explore the molecular mechanism how lycorine induced HL-60 cell apoptosis, cDNA microarray was used to investigate the expression profile of 494 apoptosis-associated genes. Real-time RT-PCR, Western blotting and immunocytochemistry were used to analyze the expression of related genes, as well as the modification and distribution of related proteins in the presence of lycorine. The results showed that 89 differential genes were expressed significantly (Cy3:Cy5 > 2 or < 0.5) among all the 494 apoptosis-related genes. 78 genes were up-regulated and 11 genes were down-regulated. We are particularly interested in the expression increase of p21 (9.271 folds) and TNF-α (8.242 folds). Furthermore, we found that lycorine could down-regulate p21-related gene expression including Cdc2, Cyclin B, Cdk2 and Cyclin E, promote the truncation of Bid protein and the release of cytochrome c from mitochondria, decrease the phosphorylation of IκB, block the nuclear import of NF-κB and down-regulate expression of survivin. This study revealed that lycorine decreased HL-60 cells survival through p21-mediated cell cycle arrest and stimulation of TNF-α signaling pathway which induced apoptosis.

Disclosures:
No relevant conflicts of interest to declare.
The Activity of AT9283 in Acute Myeloid Leukemia in Vitro and in the Clinic Is Karyotype Dependent.
Authors: Lyons John F , Squires Matthew S , Goodall John , Yule Murray , Ravandi Farhad , Foran James , Kantarjian Hagop M. 
Published: October 2019

Abstract
Abstract 4159
Aurora kinases (AK) A and B are overexpressed in a proportion of patients with acute myeloid leukemia (AML) and the level of overexpression correlates with their sensitivity to AK inhibition in vitro. We recently reported data from a dose escalation study of AT9238, a small molecule inhibitor of AKs in patients with refractory leukemias in which eight of 24 AML patients with relapsed/refractory AML achieved a 3 33% reduction in bone marrow blasts and haematological improvement. All patients had received at least one line of previous therapy. Further analysis has revealed that of the eight patients with relapsed/refractory AML that benefited from treatment with AT9283 five had a normal karyotype and the remaining three patients showed evidence of isolated abnormalities of chromosome 7, including 7q loss.
Separately, we have reported that AT9283 inhibits the proliferation and survival of AML cell lines in vitro and suggested that those cell lines with complex karyotypic abnormalities responded differently from normal diploid lines.
In these experiments AML cell lines exhibit one of two phenotypes following exposure to AT9283; rapid induction of cell death at low nM concentrations (Phenotype 1) or endo-reduplication followed by cell death at a later time point (Phenotype 2). In both scenarios treatment with AT9283 results ultimately in cell death. Cell lines with a normal karyotype tended to undergo rapid apoptosis without evidence of endoreduplication at low concentrations of AT9283. These findings provide further support for the potential importance of karyotype as a determinant of outcome in the clinical study. This is the first indication that cytogenetics might be used to predict responsiveness to Aurora kinase inhibitors in the clinic.

Disclosures:
Lyons: astex therapeutics: Employment. Squires:Astex Therapeutics, Ldt: Employment. Goodall:astex therapeutics: Employment. Yule:Astex Therapeutics Ldt: Employment. Ravandi:BMS: Consultancy, Honoraria, Research Funding.
Elderly Acute Myeloid Leukemia: A Retrospective Analysis of 93 Cases.
Authors: Du Xin , Wang Yuchun , Lu Zesheng , Weng Jianyu , Zou Xiaoli 
Published: October 2019

Abstract
Abstract 4163

Background
Acute myelogenous leukemia (AML) is generally looked upon as a heterogeneous group of different entities originating from the malignant transformation of a hematopoietic progenitor or stem cell. AML incidence increases with age. AML is associated with a poor prognosis, particularly in older patients. The poor prognosis in older adults is because of a combination of factors, including a larger proportion of cases with adverse-risk cytogenetics and other deleterious genetic and epigenetic changes, lower rates of achieving complete remission with intensive chemotherapy, higher risk of disease recurrence after achieving remission, greater comorbidity, inability to tolerate conventional allogeneic bone marrow transplantation regimens, and other adverse prognostic factors. The outlook for these patients remains dismal and little progress as been made in the last two decades. Current treatment of elderly AML consists of intensive chemotherapy with an anthracycline and a cytarabine. To evaluate our progress in the diagnosis, treatment and outcome of this condition, we reviewed the reports of 93 newly diagnosed acute myeloid leukaemia patients>/=55 years of age admitted to our department between 2003 and 2007.

Methods
A retrospective chart review was performed of 93 patients >/=55 years of age diagnosed with acute myeloid leukemia. The clinical efficacy was observed and the overall survival(OS) were analyzed.

Results
Median age of patients was 66 (range 55-88) years, 69 patients(74%) received a combination of an anthracycline and a cytarabine, either mitoxantrone per day on days 1, 2(MA),or 35 mg/m2 pirarubicin(TA) 7–8mg/m2 or idarubicin per day on days 1, 2 (IA). Each patient was given 100mg/m2 cytarabine intravenously per day for 7 days. And 24 patients (26%) were untreated. Total CR rate was 45% after 2 regimens. Only 3 patients happened to early death. The median period of the bone marrow depression is 19 days. The follow-up was ended at Aug 2008 year. Median survival of the treated patients was 256 days and it was significantly (P = 0.003) different from the untreated patients(66 days). Of the treated group, 7 patients were still alive, and the longest time was more than 6 years.

Conclusions
The prognosis of acute myeloid leukemia in older patients remains poor, but the anthracycline chemotherapy seems to improve the outcomes among older with AML. For this difficult patient cohort, it is a better treatment that can exert a more selective anti-leukaemic activity whilst not suppressing normal haemopoiesis or having significant systemic toxicities.

Disclosures:
No relevant conflicts of interest to declare.
PAKTM-Lx: A New Luminex*®-Based Assay to Detect and Identify Antibodies to Human Platelet Antigens (HPA) in Patient Serum.
Authors: Rathore Vipul , Meyers Matthew , Ray Bryan 
Published: October 2019

Abstract
Abstract 4187
Platelets express a variety of glycoproteins (GPIIb-IIIa, GPIa-IIa, GPIb/IX, GPIV, CD109 and Class I HLA). Human Platelet Antigens (HPA) represent polymorphisms of these platelet surface glycoproteins. While these polymorphisms do not alter the function of the proteins, they can become target of the immune system in mismatched individuals. Alloantibodies against HPA are involved in the pathogenesis of Neonatal Allo-Immune Thrombocytopenia (NAIT), Post-Transfusion Purpura (PTP) and refractoriness to random donor platelets. In addition to generating anti-HPA antibodies, these patients also develop antibodies to Class I Human Leukocyte Antigen (HLA). Most reference laboratories now use a combination of a sensitive screening assay such as the Platelet Immunofluorescence Test (PIFT) along with a solid phase assay for antibody identication, usually either the ELISA and Monoclonal Antibody Immobilization of Platelet Antigens (MAIPA) as well as genotyping techniques for at least HPAs 1, 2, 3, and 5. Currently, MAIPA is considered the gold standard in detection of antibodies to HPA. However, all existing methods have significant limitations in terms of sample requirements, time and labor involved. These factors are a major impediment as to how many specificities can be resolved in a rapid, efficient and reproducible manner. Development of microbead-based assays like the xMAP (Luminex®) has provided a technology that can resolve these questions. GTI Diagnostics® and Gen-Probe have developed PAKTM -Lx, a bead-based assay for the Luminex® platform to detect and determine the specificity of anti-HPA antibodies. PAKTM -Lx is capable of resolving the specificities of antibodies to HPA 1, 2, 3, 4 and 5 in addition to detecting antibodies against GPIV and HLA Class-I antigens. Furthermore, it is capable of detecting antibodies to multiple HPA and HLA with only 10 μl of sample. With a total time of less than two hours from addition of samples to end results, PAKTM -Lx is sensitive, rapid and simple to perform. Focused, flexible multiplexing in the range of 1 to 100 analytes provided by Luminex® technology meets the needs to expand the panel to include more HPA combinations, thereby reducing sample requirements, labor and costs.

Disclosures:
No relevant conflicts of interest to declare.
Comparing Efficacy of Apheresis and Whole-Blood Derived Platelets Transfusions in Acute Leukemia Patients Who Undergo Chemotherapy without Remission.
Authors: Chen Qin-fen , Li Pei , Xie Yi 
Published: October 2019

Abstract
Abstract 4189

Objective
To determine the efficacy between apheresis platelet concentrates(APCs) and platelets (PLTs) derived from whole blood(WBD) transfusion in acute leukemia patients receiving chemotherapy without remission.

Methods
All patients received same ABO group and leukocyte depletion PLTs. According to the platelet count before and after transfusion, we used percent platelet recovery(PPR) and corrected count increment(CCI) at 24 hours post-transfusion to analysis the outcomes.

Results
A total of 130 patients between the ages of 14 and 80 received 771 times PLTs transfusion. 85 patients were males, and 45 were females. There were 596 times APCs and 175 times WBD. In APCs, the platelet counts were 11.97±10.43×109/L and 24.68±18.63×109/L pre- and post-transfusion(P<0.001). In WBD, the platelet counts were 11.06±7.58×109/L and 22.88±18.00×109/L pre- and post-transfusion(P<0.001). But there was no significant difference in the platelet count between the two types of platelet produces transfusion. According to PPR, if PPR<20% is considered platelet transfusion refractoriness(PTR), the incidence rates of PTR in APCs and WBD were 46.98% and 49.71%, respectively. According to CCI, if CCI<4.5×109/L is considered PTR, the incidence rates of PTR were 42.95% and 46.86%, respectively. The incidence rate of PTR was no significant difference in PPR and CCI between the two PLTs products.

Conclusion
For the patients of acute leukemia without remission, the incidence rate of PTR was high, even if they received same ABO group and leukocyte depletion products. The efficacy was not significant difference between APCs and WBD transfusions.

Disclosures:
No relevant conflicts of interest to declare.
Poor Hematopoietic Stem Cell Mobilizers in Multiple Myeloma:A Single Institution Experience.
Authors: Ruiz-Delgado Guillermo J. , Lopez-Otero Avril , Hernandez-Arizpe Ana , Ramirez-Medina Aura , Ruiz-Arguelles Guillermo J. 
Published: October 2019

Abstract
Abstract 4237
High-dose chemotherapy rescued by autologous peripheral blood stem cell transplantation is considered standard of care for patients with multiple myeloma. Hematopoietic stem cell mobilization is accomplished by administration of hematopoietic growth factors combined or not with myelosuppressive chemotherapy. In a single institution, in a group of twenty eight myeloma patients deemed eligible for autologous transplant, stem cell mobilization was attempted using only filgrastim: Twenty six individuals were given 31 autografts employing one to four (median three) apheresis sessions, to obtain a target stem cell dose of 1 × 106 CD34 viable cells / Kg of the recipient. The median number of grafted cells was 7.56 × 106 CD34 viable cells / Kg of the recipient; the range being 0.92 to 14.8. By defining as poor mobilizers individuals in which a cell collection of less than 1 × 106 CD34 viable cells / Kg was obtained, a subset of eight poor mobilizers was identified; in two patients the autograft was aborted because of an extremely poor CD34 cell yield (less than 0.2 × 106 CD34 viable cells / Kg of the recipient) after four apheresis sessions. The long-term overall survival of the patients grafted with more than 1 × 106 CD34 viable cells / Kg was better (80% at 80 months) than those grafted with less than 1 × 106 CD34 viable cells / Kg (67% at 76 months). Methods to improve stem cell mobilization are needed and may result in obtaining better results when autografting multiple myeloma patients.

Disclosures:
No relevant conflicts of interest to declare.
Sezary Syndrome Aberrant Expression of CD26 On CD4+, CD25+, FOXP3+ T Lymphocytes and Prognostic Significance.
Authors: Corinaldesi Giorgio , Corinaldesi Christian 
Published: October 2019

Abstract
Abstract 4244
The Sezary syndrome (SS) is a variant of cutaneous lymphoma (CTLC) the cell is a malignant pleomorphic CD4+/CD25+/FOXP3+ T cell with a convolute-cerebriform nucleus, a cytogenetic evidence of an abnormal clone were observed in all patients MTAP (17p13.1-TP53), SMARCE (17q21, 1q21.3- STAT5A/STATB), DULLARD (9p21.3-CDKN2A/CDKN2B), M13930 (8q24.21-C-MYC/PLEC1), expression profiles showed upregulation of genes involved in TNF-signaling pathaway, and decreased expression of tumor suppressor genes such as TGF-beta receptor II, while TWIST and EPHA4; BCL-2, and TEX10 were overexpressed. Aberrant expression of GATA -3, TRAIL, T-plastin, CD1D are present in more advanced stages and correlate with disease progression. The most characteristic symptons include a persistent xerosis, erythematous patches, pruritus, ectropion, alopecia, dystrophic nails, palmoplantar keratoderma lymphadenopathy, hepatosplenomegaly, and sometime fungal infection or colonization with staphylococcus aureus. The loss of CD26,and a decreasing of CD4+CD7- expression,and an increasing of CD25+(alfa-chain of IL-2 receptor) FOXP3+ Tregs is a costant feature; the association with the circulating aneuploid cells showed high expression of skin manifestation, disease progression and poorest prognosis.

Disclosures:
No relevant conflicts of interest to declare.
Imatinib Increases Cytotoxicity of Melphalan and Their Combination Allows An Efficient Killing of CML Cells.
Authors: Giallongo Cesarina , Cava Piera La , Tibullo Daniele , Parrinello Nunziatina , Guagliardo Provvidenza , Fabro Vittorio Del , Stagno Fabio , Chiarenza Annalisa , Consoli Carla , Spina Paolo , Vigneri Paolo , Palumbo Giuseppe A , Di Raimondo Francesco 
Published: October 2019

Abstract
Abstract 4254
BCR/ABL-positive cells are relatively resistant to chemotherapy and, in order to evaluate the effect of Imatinib (IM) in reverting drug-resistance, we evaluated on K562 the toxicity of 1 h exposure to cytosine arabinoside (ARA-C) 20 μM, hydroxyurea (HU) 100 μM, and melphalan (MEL) 20 μM, after a pre-treatment of 24 h with 1 μM IM. The doses of the drugs were similar to that achieved in the plasma after standard chemoterapeutic treatment. Cell viability was evaluated by ATP-lite at 24, 48 and 72 hs from beginning of drug-free condition. The combinations of IM plus MEL induced the highest cytotoxicity (P<0,001 at 24, 48 and 72 hs vs MEL alone) indicating that pre-treatment with IM increased K562 exposition to the genotoxic damage of MEL. We next analyzed effects on cell cycle and DNA damage by alkaline comet assay induced by this drug combination and we observed that DNA damage peaked at 48 h with IM/MEL combination. In addition, flow cytometry analysis showed that IM/MEL combination reduced the cell accumulation in G2/M phase induced by MEL (P<0.001 vs MEL), thus reducing the ability to DNA repair and recovery.
These data indicate that inhibition of BCR/ABL activity by IM increased cell cytotoxicity of MEL by reducing the effectiveness of the DNA-repair pathways and decreasing the time for DNA repair at the G2/M checkpoint..
To ascertain that these results were linked to BCR/ABL inhibition, TonB.210, a cell line where the BCR-ABL expression is inducible by doxycycline (DOX), were treated in the same conditions. Only TonB.210 cultured with DOX were insensitive to MEL while IM/MEL combination reverted these drug resistance (P<0,001).
In the final step we studied the sequential association IM/MEL on the proliferative potential of myeloid progenitors of 6 CML patients at diagnosis. IM/MEL combination increased the reduction of the overall number of colonies in comparison to IM alone (P<0.05 vs IM). In addition, the analysis on CFU-GM and BFU-E colonies by qRT-PCR demonstrated that the IM/MEL combination led to the highest reduction in the number of BCR/ABL positive colonies (P<0.01 vs IM).
Therefore, our data indicate that the pre-inhibition of BCR/ABL activity by IM increases the toxicity of MEL and allows an efficient killing of human leukemic cells, thus suggesting new therapeutic combinations for CML patients

Disclosures:
No relevant conflicts of interest to declare.
High Incidence of Radiation Induced Cavernous Hemangioma (RICH) in Long Term Survivors Who Underwent BMT with Radiation Therapy During Childhood or Adolescence.
Authors: Koike Takashi , Yanagimachi Noriharu , Yabe Hiromasa , Yabe Miharu , Morimoto Tsuyoshi , Shimizu Takashi , Ishiguro Hiroyuki , Takakura Hiromitsu , Ando Kiyoshi , Kato Shunichi 
Published: October 2019

Abstract
Abstract 4292

INTRODUCTION
Radiation induced cavernous hemangioma (RICH) is a late complication of cerebral radiation therapy. An increased number of long term surviving blood and marrow transplantation (BMT) recipients have recovered from their primary disease but are at risk of RICH.

METHODS
We investigated 66 patients who underwent BMT during childhood or adolescence. We evaluated RICH numbers, size, location and their annual changes. Furthermore we developed scoring system of RICH in order to classify severity. MRI of the brain was performed annually for 5 to 27 years after BMT, including gradient-echo sequence (T2* weighted image). RICH SCORE 1-4 is designated as mild, 5-9 as moderate and 10 or more as severe.

RESULTS
Twenty-five patients (37.9%) was diagnosed RICH. The age at the time of the diagnosis was 11-40 years old (median 27 years old). The age at the time of BMT was 1-22 years old (median 9 years old). The period from BMT to diagnosis was 10-24 years (median 16 years). All cases received TBI as conditioning of BMT and/or cranial radiation (CR) prior to BMT as treatment of primary disease. RICH was found in 25/48 (52%) who received TBI and/or CR, and was not found in any of 18 patients without radiation therapy to the brain. Total dose to the brain was 10-36 Gy. Clinical manifestations were present only in four cases. RICH SCORE ranged 1-18 points (median 4 points). Small RICHs tended to be recognized only by T2* weighted image, but not by routine imaging methods. Classification of the severity was mild in 13 patients, moderate in 8 patients and severe in 4 patients. Severity was correlated with higher radiation dose and/or with younger ages at transplantation. RICH SCORE increased yearly in 7 of 25 patients. One case developed giant RICH more than 40mm as shown in the attached image.

CONCLUSION
High incidence of RICH was found in long term survivors who underwent BMT with radiation therapy. Since all of those patients did not show RICH before BMT and all positive patients had a history of radiation therapy to the brain, the cause of RICH in those patients was considered to be radiation. Careful and long term evaluation with MRI including T2* weighted image is necessary in BMT recipients who received radiation therapy prior and/or during BMT.

Disclosures:
Ando: Alexion: Research Funding.