bluuebunny/crossref_metadata_2025_split huggingface dataset:
Creation Of Patient Derived AML Xenografts Displaying Distinct Phenotypes and Geneotypes
Authors: Li Henry Qixiang , Liu Jingping , An Xiaoyu , Wang Na , Huang Liang , Wu Ran , Cai Jie , Wery Jean-Pierre 
Published: October 2019

Abstract
We have recently successfully engrafted leukemic cells from bone marrow of several AML patients into immunocompromised NOD/SCID mice. One of them, AM7577, was reported in the last year ASH1. This model displayed typical aggressive AML disease of M5 subtype, which starts at bone marrow and gradually expand to peripheral (spleen, lymphonode and peripheral blood…). AM7577 also harbors interesting genotypes of mutations for IDH2-R140Q, FLT3-ITD, DNMT3A R882H and NPM1.
Our second AML PDX model, AM8096, was established similarly using bone marrow from a recurrent patient with AML-M2 disease. AML8096 displayed similarly aggressive disease and the same 100% mortality, including typical symptoms (BW loss, hunched, inactivity, labored breathing, ruffled coat and eventual mortality) and with abundant leukemic cells in bone, etc. The leukemic cells can serially be passed in mice with 100% take-rate and cause consistent disease (even with < 1e4 cells). This also creates a renewable and potentially unlimited source of leukemia cells. The leukemic cells in mice are identical to those of the original patient leukemic cells (CD45+, CD33+, CD13+, CD123+, and CD19-. However, some aspects of disease presentations are vastly different from that of AM7577. First, the peripheral symptom is significant less characterized by lower leukemic counts in peripheral blood and only slightly enlarged spleen and smaller lymphonode. Second, the leukemic cell morphology is also rather different with AM8096 demonstrating less differentiated phenotype. Third, the genotype, in contrast to AM7577, is wild-types for all the above oncogenes. While we have not identified the likely leukemogenesis driver gene for this model, we are currently performing RNAseq of AM8096, along with AM7577, in order to explore the underlying molecular mechanisms that drive both diseases. In addition, we are also investigating the drug response to standard of care (SOC), as compared to those AM7577.
In summary, the two AML models could serve as useful experimental models to investigate the diverse leukemogenesis, including molecular mechanisms by genetic profiling. Ultimately, they can be used to help to identify new treatment strategies for AML.
1. Liu, e.a. A unique leukemia mouse model established from AML patient with IDH2 R140Q and FLT3-ITD mutations among other common AML mutations. ASH-2012 Annual Meeting (2012).

Disclosures:
Li: Crown Bioscience: Employment. Liu:Crown Bioscience: Employment. An:Crown Bioscience: Employment. Wu:Crown Bioscience: Employment. Cai:Crown Bioscience: Employment. Wery:Crown Bioscience: Employment.
Identification and Characterization Of a Potent FLT3 Inhibitor
Authors: Chen Yun , Guo Yao , Ho Wanting , Joe Zhizhuang 
Published: October 2019

Abstract
Acute myeloid leukemia (AML) is a malignant myeloid disorder for which there is no effective treatment. Gain-of-function mutations of tyrosine kinase FLT3 are frequently found in AML patients. This makes FLT3 an attractive therapeutic target. Currently, several potent FLT3 inhibitors have been developed. However, their clinical efficacy is limited largely due to their poor effectiveness toward the FLT3-D835 mutants which are often present in AML or acquired after treatment of FLT3-ITD-positive AML with tyrosine kinase inhibitors. Needless to say, more potent FLT3 inhibitors targeting both FLT-ITD and FLT3-D835 mutants are needed. In addition, combinations of tyrosine kinase inhibitors with drugs targeting other signaling pathways represent a new trend in anti-cancer drug development.
To establish an effective kinase assay for FLT3 inhibitor screening, we generated a protein substrate designated GST-FLT3S which was expressed in E. coli cells as a glutathione S-transferase fusion protein. The protein substrate together with recombinant proteins containing the catalytic domain of wild type and mutant forms of FLT3 expressed in baculovirus was used in biochemical screening of inhibitors. Several potent inhibitors were obtained. Importantly, one of the inhibitors with an oxindole core structure inhibited FLT3 and D835 FLT3 mutants equally well with nanomolar IC50 values. We further analyzed the potency of the inhibitor by performing cell-based assays. The cells used included FLT3-ITD-positive cell line MV-4-11 and an EPO-dependent erythroleukemia cell line transformed by retrovirus mediated expressions of FLT3-ITD and FLT3-D835 mutants. At nanomolar concentrations, the inhibitor blocked growth factor signaling and effectively caused apoptosis and cell cycle arrest. It showed significant advantage over the current available FLT3 inhibitor, sorafenib.
Loss-of-function mutations of tumor suppressor p53 are common in solid tumors but relatively rare in AML although its expression is often suppressed. This makes p53 a potential target for anti-AML drug development. We employed MDM2 inhibitor nutlin-3 which blocks the degradation of p53. Importantly, at sub-nanomolar concentrations, FLT3 inhibitors and nutlin-3 synergistically inhibited growth of cells containing FLT3-ITD or FLT3-D835 mutants.
Altogether, we developed an effective substrate for screening of FLT3 inhibitors and identified one compound with high potency toward both FLT3-ITD and FLT3-D835. We further demonstrated that targeting FLT3 and p53 simultaneously greatly increases drug potency.

Disclosures:
No relevant conflicts of interest to declare.
The Survival Of The Elderly Acute Myeloid Leukemia Patients Do Not Depend On The Intensity Of Induction Chemotherapy. Retrospective Study Results
Authors: Grosicki Sebastian , Bodzenta Ewa , Kriegler Marek , Szypula Ilona , Fejklowicz Marcin , Twardosz Maja , Kurzawa Teresa , Barchnicka Agnieszka , Melisz Aleksandra 
Published: October 2019

Abstract
The aims of this study were comparison of overall survival (OS) and complete remission rate (CR) in whole group of acute myeloid leukemia (AML) >60 patients, and in younger older (61 – 70) AML patients in dependency of qualification to risk group according to PALG stratification and a schedule of treatment.
Patients with AML >60 after diagnosis were qualified into the risk groups according to PALG stratification for AML >60. The patients from group 1 patients were receiving intensive induction chemotherapy DAC (daunorubicin, cytarabine, cladribine) or DA (daunorubicin, cytarabine), those from group 2 were receiving cytarabine+thioguanine induction chemotherapy, and those from group 3 chemotherapy with low doses of cytarabine. Patients from first group, who reached CR after induction were receiving consolidation with mitoxantrone + standard dose of cytarabine, and next they were qualified to HSCT or maintenance phase. Patients from group 2 and 3 independently of results of induction were receiving successive cycles of chemotherapy like in first course every 4 weeks to 2 years with very intensive supportive care.
One hundred sixteen patients with untreated AML treated In period 02.2009-12.2012, age 73 (61-89), men 50% were included to this study. Thirty three patients were qualified to group 1, 50 to second and 33 to third. After preliminary analysis, because of similar early and late outcomes of the therapy in group 2 and 3, those groups of patients were joint for next analysis.

Disclosures:
No relevant conflicts of interest to declare.
Outcome Of Dose-Adjusted Decitabine Regimen Compared With FLAG Regimen In The Treatment Of Relapsed/Refractory Acute Myeloid Leukemia
Authors: Wu Qian , He Guangsheng , Depei Wu , Sun Aining , Qiu Huiying , Jin Zhengming , Miao Miao , Tang XiaoWen , Han Yue , Shen Yiming , Chen Feng , Hu Xiaohui , Jin Song , Zhang Xuhui 
Published: October 2019

Abstract

Background
The salvage therapy in patients with relapsed/refractory acute myeloid leukemia still poses a highly unmet clinical need. Given the established activity and toxicity profiles of hypomethylating agents such as 5-aza-2'-deoxycytidine (decitabine, DAC) in patients with untreated acute myeloid leukemia (AML), we explored the therapeutic effects of dose adjusted DAC in patients with relapsed / refractory AML, and compared treatment outcome with the conventional FLAG regimen.

Patients and Methods
27 patients with relapsed/refractory AML were included in this analysis. Twelve patients were treated with DAC 20 mg/m2 per day as 1-hour intravenous infusion for consecutive five days, with additional 1-3 doses of DAC added based on response and tolerability (Table 1). Another group of 15 patients received a course of FLAG regimen as controls.

Results
Although the overall response rates (ORR) were similar in DAC group (5/12) and FLAG group (9/15)£¨41.7 % versus 60 %, P=0.449£©, the complete remission (CR) rate plus CRi was lower in DAC group (2/12) than in FLAG group(10/15)£¨16.7 %versus 66.7 %, P=0.047£©. Induction mortality was 0 (0% at 8 weeks) and toxicities were manageable in both groups. Toxicities were predominantly hematologic. The most common drug-related adverse events (AEs£© were grade 4 myelosuppression which were comparable for DAC and FLAG., DAC group hadfewer infections£¨DAC, 6/12£»FLAG£€n=14/15, P=0.024£©. The 2-year survival rate was similar in the two groups: 25.0% in the DAC versus 26.7% in the FLAG group£¨P=0.574£©, while median survival times of the two groups were 4 and 12 months, respectively.

Conclusion
Dose-adjusted DAC achieved similar overall response rate with lower infection risk compared to FLAG regimen in patients with relapsed / refractory AML.

Disclosures:
No relevant conflicts of interest to declare.
Acute Myeloid Leukemia In Patient Over 90 Years Old: Where Do We Go From Here? Case Report
Authors: Bellesso Marcelo , Dias Daniela Ferreira , Centrone Renato , Santucci Rodrigo , Nicodemo Izabel Pernambuco 
Published: October 2019

Abstract

Introduction
Acute Myeloid Leukemia (AML) widely affects elderly patients, with disappointing survival rates with increasing age. Although chronological age is an independent prognostic risk, it is really important to understand that this group is heterogeneous, so a geriatric assessment may be helpful before making decisions regarding therapy. We describe a 91 year-old patient with AML treated with decitabine, achieving a complete response and good quality of life for 10 months.

Case Report
In May 2012, a 91 year-old woman with myelodysplastic syndrome was admitted at the emergency department presenting asthenia, pallor, pain and edema of the left inferior limb. Her blood count showed: hemoglobin 10.7g/dL, leukocytes 81.800/mm³ with 95% blast cells, and platelets 45.000/mm³. In addition, a Doppler ultrasound evidenced deep venous thrombosis in her left leg. A bone marrow aspirate confirmed AML with myelodysplasia-related alterations, with 95% blast cells, with positive expression of CD45, CD33, MPO, and CD117 (CD45+; CD33+; MPO+; CD117+). Her karyotype was 46,XX [20 cells analyzed]. She was first treated with hydroxyurea. Afterwards, as her performance status improved and in spite of her age, we decided to treat her with Decitabine 20mg/m²/day for 5 days, because she had no severe comorbidity or any severe impairment of every-day-life instrumental activities. After a first cycle with grade 4 neutropenia and thrombocytopenia, she was discharged. After 40 days, complete hematologic recovery was observed. She received seven treatment cycles, and the most important symptom of toxicity was febrile grade 3 neutropenia in the 3rd cycle, with the only inpatient treatment. After this intercurrence, the doses were reduced by 25%. The patient achieved complete remission, spent eleven months without needing blood transfusions and with an acceptable quality of life, but then she relapsed, presenting persistent neutropenia and 44% of blasts in a bone marrow aspirate (05/27/2013). She died on 07/02/2013 due to the progression of the disease.

Conclusion
This case shows that, despite her advanced chronological age, our patient, affected by a fatal disease, lived approximately 13 months with optimal response to treatment (with Decitabine) and enjoying an acceptable quality of life for ten months. Considering this patient profile, could we conclude that this outcome is the best our treatment can aim to achieve?

Disclosures:
No relevant conflicts of interest to declare.
Disulfiram, an clinically used anti-alcoholism drug has the potential to target acute myeloid leukemia cells in a copper-dependent manner in vivo
Authors: Xu Bing , Li Rongwei , Dong Huijuan , Chen Feili , Liu Yuejian , Wang Shiyun , Zhou Shuyun 
Published: October 2019

Abstract

Background
Disulfiram(DS), an old drug clinically used for alcoholism, was reported to have antitumor effects, recent studies have found that Copper(Cu) can significantly enhance the DS-induced cell death in vitro in a variety of tumor cells. Our previous studies also demonstrated that disulfiram/copper (DS/Cu) couldtarget human leukemia cell lines(like KG1α,Molt4) through the activation of JNK, in vitro. However, there is few report about the ability of DS/Cu in killing cancer cells in vivo.

Aims
This study aims to explore the effect of DS/Cu on acute myeloid leukemia cell line KG1αin vivo and clarify the underlining mechanism.

Methods
6-8 week old female NOD/SCID mice were sublethally irradiated with 2Gy X-ray the day before transplantation, followed by intravenous injection of KG1α cells (1×107 cells) suspended in 0.2 mL of PBS. 5 weeks after transplantation mice were randomly divided into three treatment groups: vehicle (0.9% saline), a combination of DS and Cu daily for 2 weeks, Ara-C alone twice before killing. Mice were sacrificed after 2 weeks treatment with tissues of spleen, liver, bone marrow being observed using histopathology method to detect the invasion of leukemia. The DS/Cu-induced p-c-jun activation was also examined by western blot using tissues of spleen, liver, bone marrow. Statistical analysis was carried out with one-way ANOVA to assess statistical significance (*p < 0.05).

Results
4 weeks after transplantation, mice were dispirited with low appetite, down-bent gait, wrinkled fur, slow move, just like suffered from leukemia. What’s more, immature blasts like morphology similar to KG1α were found in the peripheral blood of the mice(11%±3.41). All the mice were sacrificed after 2 weeks treatment, mice in control group were observed with slightly larger spleen and liver with the morphology of invasion of leukemia such as a granular appearance than the other two groups. Histopathology examination showed that leukemia cells infiltrate liver, spleen and bone marrow, and the immunohistochemistry examination found that the leukemia cells in spleen, liver and bone marrow expressed human specific antigen CD45 with the highest expression level in the control group. Moreover, solid tumor could be observed in the peritoneal cavity of two mice in the control group with expression of human specific antigen CD45detected by immunohistochemistry examination. Western blot in this study showed DS/Cu complex induced phosphorylation of c-Jun expression in the spleen, liver and bone marrow.

Conclusion
DS/Cu complex could effectively target the acute myeloid leukemia cells in the acute leukemia NOD/SCID mice while inhibiting the invasion of leukemia to some extent, and the activation of JNK might play a functional role in DS/Cu mediated antileukemic effects.

Disclosures:
No relevant conflicts of interest to declare.
PET Scan Negativity Post Treatment In DLBCL May Not Indicate A Cure
Authors: Atherton David , Lachica Mila , Kuo Yen-Hong , Topilow Arthur A. 
Published: October 2019

Abstract

Background
Diffuse Large B-Cell Lymphoma (DLBCL) is an aggressive Non-Hodgkin’s lymphoma. Previous work has suggested that a negative FDG-PET/CT scan, following chemotherapy with curative intent, indicated a cure. However, the understanding that a negative PET post-treatment indicates that no further PET scans are needed, has not yet been accepted as a standard of care. We reviewed the medical records of DLBCL patients with negative post-treatment PET scans in order to evaluate if further PET scanning is superfluous and that the treatment has effected a cure.

Methods
A list of DLBCL patients diagnosed and treated at JSUMC from 2000 to 2010 was obtained from the Tumor Registry. Patient health information was reviewed using electronic as well as paper records.

Results
A total of 133 DLBCL patient charts were reviewed and only 40 had complete PET scan records available at the time of study. Thirteen of the 40 fit the criteria of a negative post-treatment PET scan. Sixty-nine percent (9/13) were males and 31% (4/13) were females. The average age at diagnosis was 61 years. Seventy-seven percent (10/13) had a complete response and did not relapse based on their subsequent PET scan reports. Twenty-three percent relapsed after 14, 22, and 25 months, respectively (3/13, 95% confidence interval: [5%, 54%]). The mean (standard deviation) annual number of PET scans was 1.65 (0.68). The frequency ranged from 0.55/year to 2.67/year.

Conclusions
Three of 13 DLBCL patients eventually relapsed after a negative post-treatment PET scan. This study suggests that careful monitoring of DLBCL patients who have negative PET scans is needed after treatment. Further studies need to be done to confirm the time period a PET scan may be most effective in restaging the disease.

Disclosures:
No relevant conflicts of interest to declare.
The Research Of TCR Vβ Clonality Distribution In Patients With Diffuse Large B-Cell Lymphoma and Its Clinical Significance
Authors: Wang Chunyan , Tan Huo , He Ganlin , Li Yangqiu 
Published: October 2019

Abstract

Background
This study investigated the gene expression of TCR Vβ subfamily T cells and clonality in peripheral blood from patients with DLBCL and discussed the relation between TCR VB subfamily and clinical characters, prognosis, T Cell Subgroup, et al. this study include 14 patients with DLBCL, to analyzed TCR Vβclonality of their blood samples and to comprehend the CDR3 sequence.

Methods
The complementarity determining region 3(CDR3)of TCR Vβ subfamily genes in peripheral blood mononuclear cels(PBMCs) from 14 DLBCL patients were amplified using RT-PCR, and the products of PCR were further analyzed by genescan technique for detection of the CDR3 size,to evaluate clonality of TCR Vβ T cells.
After that, analyzing the data with statistics software SPSS 17.0, to find out the relationshop between clonality of TCR Vβ(oligoclon, oligoclone trendenc, monoclonal bioclone or polyclon ) and laboratory features, immunophenotype, period, efficacy of chemotherapy, prognosis, et al.

Result
This study magnified samples and comprehensively analized characteristics of TCR Vβ subfamily in patients with DLBCL patients, confirming the unbalance distribution and clonal expansion of TCR Vβ subfamily. Several cases showed clonal proliferation in Vβ3, Vβ6, Vβ7, Vβ9, Vβ14, Vβ15 and Vβ22, especially in Vβ15 and Vβ22.These clonal proliferation and deletion of Vβ16, Vβ18, Vβ24 may be considered DLBCL related. Several TCR Vβ subfamily appeared 3-polyclonal. Through number of clonal proliferation, gender, age, time since diagnosis, LDH, germinal center type or nongerminal center type, period, autologous transplantation or not, initial treatment / relapse of 14 cases were analized using two independent samples t-test. initial treatment / relapse was statistically significant (p = 0.027): number of clonal proliferation in patients with relapse-DLBCL compared to the number of untreated patients is high than patients with initial treatment. Pearson correlation analysis illustrated the number of clonal proliferation and ki-67 was negatively correlated, while CD3+ CD8+ lymphocytes showed a positive correlation. for the first time discriminant analysis was applied to gene scan results in the patients with DLBCL.

Conclusion
This study confirmed the unbalance distribution and clonal expansion of TCR Vβ subfamily in patients with DLBCL. Further studies may pay attention to Vβ15, Vβ22 and 3-polyclonal which may relate to DLBCL. Several indicators of TCR Vβ subfamily are closely linked with clinical data, that TCR Vβ may be considered to use in clinical diagnosis and prognosis. It’s the first time that Discriminant analysis was applied to gene scan results, so discriminant analysis may be considered to clinical application after perfecting.

Disclosures:
No relevant conflicts of interest to declare.
Bortezomib plus Dexamethasone versus Fludarabine plus Cyclophosphamide in initial treatment of patients with Waldenström macroglobulinemia
Authors: Zhang Yaping , Lin Zenghua , Wang Xinfeng , Cao Xin , Song Guoqi , Liu Hong 
Published: October 2019

Abstract

Purpose
We examined the efficacy of bortezomib plus dexamethasone(BD) versus fludarabine plus cyclophosphamide(FC) in patients with symptomatic, untreated Waldenström macroglobulinemia(WM). Furthermore, to evaluate the toxicity of BD regimen in WM. Patients and methods: 17 untreated WM patients received either bortezomib and dexamethasone(n=7) or fludarabine and cyclophosphamide(n=10). BD group: A cycle of therapy consisted of bortezomib 1.3 mg/m2 intravenously; dexamethasone 40 mg on days 1, 4, 8, and 11. Patients received 4 cycles at least. FC group: A cycle of therapy consisted of fludarabine 25 mg/m2 on days 1-3; cyclophosphamide 250mg/m2 on days 1-3. Patients received 4-6 consecutive cycles for induction therapy. Responses were based on paraprotein levels. Herpes zoster prophylaxis was instituted with using oral valacyclovir. Results: In BD and FC groups, the overall response rates(PR+MR) were 86% versus 70% respectively. There was not complete response in two groups. In BD and FC groups, minor and partial responses occurred at a median of 25 vs. 54 days and 55 vs. 94 days respectively(P<0.05, respectively). The median progression-free survival(PFS) was 27 months, and a median follow-up of 62 months the actuarial 5-year overall survival(OS) rate was 80% in FC group. Because of a short following time, there were not PFS and OS that can be observed in BD group. Hemoglobin and platelet levels increased normally in 83% vs. 50% and 100% vs. 66% patients in BD and FC groups respectively(P<0.05, respectively). In BD group, peripheral neuropathy was the most common toxicity(57% grade 1- 2, no grade 3- 4). Hematologic toxicities included grade 3 to 4 thrombocytopenia in 2/7(28%) and neutropenia in 1/7(14%). 42% patients developed herpes zoster. Conclusions: The results demonstrate that BD produces rapid responses, along with high rates of response in WM, and it was generally well-tolerated in our study. Herpes zoster prophylaxis is necessary with BD, and peripheral neuropathy was the most common toxicity, but not leading to discontinuance of BD. This retrospective analysis confirms that BD therapy is an effective initial treatment in WM.

Disclosures:
No relevant conflicts of interest to declare.