bluuebunny/crossref_metadata_2025 huggingface dataset:
Src Family Kinase Inhibitor PP2 Enhances Differentiation Of Acute Promyelocytic Leukemia Cell Line Induced By Combination Of ATRA and Arsenic Trioxide
Authors: Kim Kyoung Ha , Cheong Hee-Jeong , Kim Sook-Ja , Yoon Jina , Kim Han Jo , Kim Se Hyung , Kim Hyun Jung , Lee Sang-Cheol , Bae Sang Byung , Kim Chan-Kyu , Lee Namsu , Lee Kyu Taeg , Park Sung Kyu , Hong Dae Sik , Park Hee Sook , Won Jong-Ho
Published: October 2019

All-trans-retinoic acid (ATRA) and arsenic trioxide (ATO) combination yields a high quality remission and survival in newly diagnosed acute promyelocytic leukemia. For subsequent similar data, NCCN guidelines indicate that ATRA plus ATO is an alternative for patients who cannot tolerate anthracycline therapy. We demonstrated that SFK (Src Family Kinase) inhibitor PP2 enhanced acute promyelocytic leukemia (APL) cell differentiation when combined with either ATRA or ATO with difference in activation of RA-induced genes. In this study, we investigated SFK inhibitor PP2 could enhances the differentiation of NB4 APL cells when combined with ATRA and ATO and the changes in the expression of intercellular adhesion molecule-1 (ICAM-1) derived from the retinoic acid receptor (RAR) target gene.

Treatment of NB4 cells with 1 nM of ATRA, 0.5 uM of ATO, or 10 uM of PP2 for 72 hours induced expression of CD11b-positive cells by 13.01%, 11.53% or 13.28%, respectively. However, the combination of ATRA and ATO and the combination of three agents (ATRA, ATO, and PP2) led to a significant higher expression of CD11b-positive cells (30.96% and 63.17%, respectively). The synergistic effect of the combination of three agents was more significant than the combination of ATRA and ATO. These results were confirmed by NBT staining. These effects were not related with apoptosis. Annexin-V-fluorescene staining revealed that combination of ATRA and ATO and combination of three agents did not induced apoptosis in NB4 cells. The expression of ICAM-1 was markedly increased in cells treated with the combination of three agents.

These findings suggest that the SFK inhibitor can enhances differentiation of APL cells combined with ATRA and ATO. FDA approved SFK inhibitors, such as dasatinib and bosutinib, may be beneficial for the treatment of APL in combination with ATRA and ATO.

Disclosures:

No relevant conflicts of interest to declare.
Induced Apoptosis Of Multidrug Resistance Leukemia K562/A02 Cells By Magnetic Nanoparticle Of Fe3O4 Copolymerizating JNK1 shRNA Expression Vector and Daunorubicin
Authors: Zhang Wei , Chen Baoan , He Xiangfeng , Fu Rong
Published: October 2019

Currently, serious toxicity and poor antitumor efficacy resulting from the induction of multidrug resistance (MDR) in hematological malignancies and solid tumors are common hindrances to successful systemic chemotherapy. One of the important mechanisms of tumorigenesis and resistance to anticancer drugs was blockade of the apoptosis-inducing pathway. In order to overcome tumor MDR while maintaining good antitumor activity, biomaterial combine with gene therapy provides a novel strategy for the treatment of MDR.The aim of this study was to explore the induction of apoptosis by magnetic nanoparticle of Fe3O4 (MNPs-Fe3O4), a novel delivery system for chemotherapeutic regimen that is chemically stable, environmentally friendly, and noncytotoxic, copolymerizating JNK1shRNA expression vector with DNR, which could overcome siRNA technique instability and enhance DNR chemotherapeutic efficiency and provide a new field to reverse tumor MDR. JNK1shRNA expression vector was constructed and screened the most effective plasmid and transfected the cells. Typical apoptotic characteristics and apoptosis effect of MNPs-Fe3O4 and PGM-3 with DNR were investigated by DAPI staining and FCM assay, respectively. Additionally, drug accumulation and apoptosis relative-genes were evaluated in K562/AO2 leukemia cells.The results showed that PGCsilencer-U6-neo-GFP-GV102 shRNA/JNK1 was successfully constructed, which was confirmed by sequencing that expression of JNK1 gene in mRNA and protein level that had the less in PGM-3 than that of PGM-1 or PGM-2. Meanwhile, MNPs-Fe3O4 and PGM-3 with DNR (DNR/PGM-3/MNPs-Fe3O4) could synergistically induced typical apoptotic characteristics of chromatin condensation and apoptotic body with DAPI staining, and a higher rate of apoptosis were detected in K562/A02 cells treated with DNR/PGM-3/MNPs-Fe3O4. Intracellular DNR was higher in K562/A02 cells treated with DNR/PGM-3/MNPs-Fe3O4 than that of DNR/MNPs-Fe3O4 by FCM analysis.  Further study demonstrated that both DNR/PGM-3/MNPs-Fe3O4 and DNR/PGM-3 reduced the gene transcriptions and protein expressions of bcl-2, enhanced that of bax and caspase-3, decreased survivin in protein level, and has no different change in mRNA level by qRT-PCR and Western blotting analyses, respectively. These findings showed that the combination of MNPs-Fe3O4 with JNK1 shRNA expression vector and DNR could induce apoptosis of K562/A02 cells through elevating the ratio of bax/bcl-2, activating caspase-3, and decreasing survivin in protein level.

Disclosures:

No relevant conflicts of interest to declare.
CUDC-907, a Dual HDAC and PI3K Inhibitor, Potentially Targets Cancer Cells and The Microenvironment In Hematological Malignancies
Authors: Wang Jing , Pursell Natalie , Ma Anna , Atoyan Ruzanna , Samson Maria , Borek Mylissa , DellaRocca Steven , Yin Ling , Wang Da-gong , Zifcak Brian , Xu Guang-xin , Voi Maurizio , Lai Cheng-Jung
Published: October 2019

Histone deacetylase (HDAC) and phosphatidylinositol 3-kinase (PI3K) inhibitors have each emerged as promising anti-cancer therapeutics that may target not only cancer cells but also their microenvironment. We have developed a strategy to simultaneously inhibit HDAC- and PI3K-dependent signaling with a single small molecule, CUDC-907. In this study, we report that CUDC-907 induces apoptosis and G2/M arrest in hematological cancer cells. CUDC-907 also impairs cytokine and chemokine production, in association with inhibition of AKT and STAT phosphorylation. Moreover, we demonstrate that CUDC-907 reduces hypoxia-induced HIF-1a expression in cancer cells and VEGF-induced tube formation in endothelial cells, suggesting potential anti-angiogenic activity. With its integrated HDAC and PI3K inhibitory activity, CUDC-907 may thus offer improved therapeutic benefit through simultaneous suppression of cancer cell proliferation and perturbation of their protective microenvironment. These studies provide support for the clinical development of CUDC-907 in hematological malignancies. A Phase I study in patients with lymphoma or multiple myeloma is ongoing.

Disclosures:

Wang: Curis Inc.: Employment. Pursell:Curis Inc.: Employment. Ma:Curis Inc.: Employment. Atoyan:Curis Inc.: Employment. Samson:Curis Inc.: Employment. Borek:Curis Inc.: Employment. DellaRocca:Curis Inc.: Employment. Yin:Curis Inc.: Employment. Wang:Curis Inc.: Employment. Zifcak:Curis Inc.: Employment. Xu:Curis Inc.: Employment. Voi:Curis Inc.: Employment. Lai:Curis Inc.: Employment.
Gene Expression Levels Of Imatinib Transporters and Molecular Response To Imatinib Therapy In Chronic Myeloid Leukemia Patients
Authors: Malhotra Hemant , Sharma Pratibha , Bhargava Shipra , Malhotra Bharti , Kumar Madhu
Published: October 2019

Imatinib mesylate (IM) is the standard first-line treatment for most CML patients. After an initial response, approximately 30 to 40% patients develop resistance to the drug. Various mechanisms of resistance to Imatinib therapy have been identified. One of the mechanisms proposed is varying expression levels of the drug transporters. In the present study, we determined the relative expression levels of Imatinib transporter genes (hOCT1, ABCB1, ABCG2) in CML patients by quantitative real time polymerase chain reaction (qRT-PCR) and correlated these levels with molecular response.

One hundred and ten CML patients were considered for gene expression analysis study for hOCT1 gene and eighty seven CML patients were considered for gene expression analysis study for ABCB1 and ABCG2 genes. CML patients who were on IM therapy for more than 2 years were divided into two groups: Responders: patients who achieve a Complete Molecular response (CMR) or a Major Molecular Response (MMR) [bcr/abl: abl ratio <1% as assessed by RQ-PCR] and Non-responders: those without CMR or MMR (bcr/abl: abl ratio =/> 1% as assessed by RQ-PCR). The relative transcript expression levels of the three genes were compared between responders and non-responders. No significant difference in the expression levels of hOCT1, ABCB1 and ABCG2 was found between the two categories - responders versus non-responders (p value > 0.05). The median transcript expression levels of hOCT1, ABCB1 and ABCG2 genes in responders were 30.63, 10.14 and 0.59 versus 40.13, 8.34 and 0.53 in non-responders, respectively.

We conclude that, in our study, the mRNA expression levels of IM transporter genes did no correlate with molecular response in CML patients.

Disclosures:

No relevant conflicts of interest to declare.
Is Superinstability Of Karyotype Of Acute Myeloid Leukemia “Relic Radiation From Big Bang” Of Chromosome Homeostasis On MDS Stage?
Authors: Luchinin Alexander S. , Turkina Anna G. , Ovsepyan Vanik A.
Published: October 2019

Introduction

In some cases of the cytogenetic analysis patients with acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) there are increase frequency non-clonal disorders (“cytogenetic noise”)  in combination with complex (≥ 3 chromosomal aberration) clonal disorders. It is not infrequently manifested in the form of superinstability of the karyotype. In this case every aberrant cell is genetic different from each other. It is obvious that super unstable karyotype has a poor prognosis even more than complex clonal disorders, since due to the high genetic instability is a strong possibility of mutations that cause resistance to chemotherapy. In other words, the high level of “cytogenetic noise” on background of complex clonal disorders can be used as a marker of highly aggressive course of AML and MDS. The aim of our study was to determine the effect of superinstability of the karyotype on the current and prognosis of AML and MDS.

Methods

Thus, 157 patients with cytogenetic survey from register of AML and MDS by Kirov Research Institute of Heamatology were included in our research. They are 73 men, 84 women have the age from 21 to 81 (Me 55) years. Among them 34 patients have MDS and 123 – AML excluding promyelocytic leukemia. Chromosomal aberration were determined from bone marrow aspirate by standard cytogenetic method (GTG-method). Was evaluated at least 20 metaphase cells.

Results

Among MDS patients the prevalence of superinstability of the karyotype was 29%, while among AML – only 4%, χ2=16.9, p<0.0001. In MDS group 2 patients (20%) are RAEB-1, 7 (70%) – RAEB-2, 1 (10%) – unspecified MDS. In AML group 2 patients (40%) are M2 FAB, 1 (20%) – M4 FAB, 2 (40%) - unspecified AML. Given that this type of cytogenetic of high instability is a characteristic feature of MDS, we can suppose, that AML with such instability always have the preliminary MDS stage with cell superinstability of the karyotype. In other words, the superinstability of the karyotype in AML can be regarded as “relic radiation from Big Bang” of chromosome homeostasis, which occurred at the preliminary MDS stage. The median of overall survival among all these patients treated by standard chemotherapy was 3 month, maximum survival – 16 month. The median of overall survival in AML patients was 1 month, MDS – 3.5 months. All patients were died.

Conclusion

A chromosomal superinstability of tumor cells that greatly enhances the aggressiveness of the current AML and MDS, leading to chemotherapy resistance and dramatically reduces the overall survival patients. Superinstability of the karyotype determines the group of patients untreatable by standard chemotherapy. The allogeneic bone marrow transplantation need in these cases.

Disclosures:

No relevant conflicts of interest to declare.
Outcome Of Patients With Acute Myeloid Leukemia/High Risk MDS According To The Kidney Function
Authors: Malkawi Ahmed , Anand Ankit , Al-Ameri Ali , Abdelfatah Mohamed , Kanaan Zeyad , Haller Nairmeen
Published: October 2019

Background

Acute renal failure or injury is a common complication of treatment of patients with acute myelogenous leukemia (AML) or high risk MDS, but the effect of renal function of patients who have acute myeloid leukemia/high risk MDS is not clearly highlighted as a predictor of survival, to the best of our knowledge this issue has not been studied in depth before.

Aim

study the effect of chronic kidney disease on the survival of patient with acute myeloid leukemia/High Risk MDS.

Methods

A retrospective study of all AML & high risk MDS patients treated at AGMC, Ohio, USA during 2001-2010. After IRB approval of the project, patients’ charts were reviewed to gather information on demographics, diagnosis types/subtypes, glomerular filtration rate (GFR), treatment, and cytogenetics. Patients were classified as low-intermediate risk or high risk according to cytogenetic background using WHO criteria. Also according to GFR patients were classified to GFR <30, 30 - 60 and > 60. Overall survival (OS) rates were determined by Kaplan-Meier Survival Analysis. Prognostic factors were evaluated by Log Rank analysis.

Result

Out of 130 patients we were able to classify 99 patients (75%). Patient were grouped into 59 Pts with GFR>60, 37 Pts with GFR 30-60 and 3 Pts with GFR<30. Time to event survival analysis was done.

Conclusion

Glomerular filtration rate GFR is a major identified factor in patients survival who have acute myeloid leukemia AML/High Risk MDS, those patients with GFR 30-60 do better in term of survival, we don’t have any explanation for that, more data with high number of patients needed to elaborate on this issue.

Disclosures:

No relevant conflicts of interest to declare.
Increased 14-3-3ζ Expression In The Multidrug-Resistant Leukemia Cell Line HL-60/Vcr As Compared To HL-60 Cell Line Mediates Cell Growth and Apoptosis
Authors: Liang Rong , Chen Xiequn qun , Bai Qing xian , Wang Zhe , Yang Lan , Zhang Tao , Dong Bao xian , Gao Guang xun , Gu Hong tao , Zhang Liang
Published: October 2019

Objectives

Acute myeloid leukemia (AML) recurrence is largely a result of multidrug resistance (MDR). Because vincristine-resistant HL-60 cells (HL-60/VCR) express greater levels of 14-3-3¦Æ, which is associated with apoptosis and chemosensitivity in other cancers, this study sought to examine its role in AML chemosensitivity using HL-60 and HL-60/VCR cells.

Methods

The mRNA and protein expression of 14-3-3¦Æ, mdr1, Pgp, BCL-2 and MCL-1 were examined using semi-quantitative RT-PCR and Western blot analyses, respectively. The subcellular location of 14-3-3¦Æ protein in HL-60 and HL-60/VCR cells was determined using immunofluorescence and confocal microscopy. After siRNA-mediated silencing of 14-3-3¦Æ in HL-60 AND HL-60/VCR cells, cell growth and cell cycle progression were determined by direct counting and flow cytometry, respectively. The effect of 14-3-3¦Æ siRNA on topotecan (TPT)-induced apoptosis was evaluated using acridine orange/ethidium bromide and Annexin V staining as well as TUNEL analysis. NF-¦ÊB-DNA biding was also assessed using electrophoretic mobility shift assay.

Results

As compared to HL-60 cells, increased 14-3-3¦Æ mRNA and protein expression was observed in HL-60/VCR cells. In addition, increased mdr-1 mRNA as well as Pgp, Bcl-2, and Mcl-1 protein expression were observed in HL-60/VCR cells. In both HL-60 and HL-60/VCR cells, 14-3-3¦Æ was observed in the cytoplasm and nuclear compartments. 14-3-3¦Æ siRNA significantly reduced HL-60 and HL-60/VCR cell growth after 48 h and increased the proportion of cells in the G0/G1 phase. Moreover, 14-3-3¦Æ siRNA significantly increased the sensitivity of both HL-60 and HL-60/VCR cells to TPT possibly through inhibition of Bcl-2, Mcl-1 and Pgp protein expression. Conversely, increased Bad and Noxa protein expression was observed with 14-3-3¦Æ siRNA. NF-ĸB DNA binding was reduced with 14-3-3¦Æ siRNA.

Conclusions

Silencing of 14-3-3¦Æ increased the sensitivity of both sensitive and resistant HL-60 cells to TPT-induced apoptosis possibly through altering the expression of apoptosis-associated proteins, suggesting that it may be a potential target for MDR AML.

Disclosures:

No relevant conflicts of interest to declare.
Single Arm, Multi-Center, Phase II Study Of Clofarabine In Chinese Pediatric Patients With Refractory Or Relapsed Acute Lymphoblastic Leukemia
Authors: Zhang Leping , Du Xin , Li Yan , Cai Zhen , Yu Kang , Tan Fenlai , Fang Yi , Huang Xiaojun , Lu Jin
Published: October 2019

Background

Clofarabine is a second generation purine nucleoside which inhibits DNA polymerase alpha and ribonucleotide reductase, leading to the depletion of intracellular deoxynucleotide triphosphate pools, disruption of mitochondrial function and apoptosis. A phase II study was conducted to evaluate the safety and efficacy of clofarabine in pediatric patients with refractory or relapsed acute lymphoblastic leukemia (ALL) in China.

Method

Patients aged 1-21 years with histologically confirmed, relapsed and/or refractory ALL were eligible. Clofarabine was administered intravenously for 5 days at 52 mg/m2/d during induction and consolidation. The primary end point was overall remission rate (ORR; complete remission [CR] plus CR with incomplete counts [CRi]). Blood samples were obtained from patients who agreed to blood drawing for pharmacokinetic determinations.

Results

A total of 44 patients were treated in this study, in which 43 were evaluable for response. The median age was 13 years (range: 5 to 22 years), and the majority were B cell-ALL (75%, 33/44). The overall remission rate was 44.2% (19/43), including two complete remissions (CR), and 17 CRis. 39 patients had progressive disease at cut-off time, the overall median remission duration was 2.9 months, and the median remission duration was 3.4 months for patients achieved CR or CRi. Seven CR/CRi patients had chance to got HSCT after clofarabine treatment. Subgroup analysis showed that patients younger than 14 years and with B cell-ALL had a higher ORR than patients older than 14 years and with T cell-ALL, and the difference were statistically significant (age < 14 years vs. age ≥14 years: 60.9% vs. 25%, p =0.0308; B cell-ALL vs. T cell-ALL: 57.6% vs. 0, p=0.002). The most common (≥10%) adverse reactions (ADR) were hematological toxicity, gastrointestinal reaction, and abnormal liver function, and the most common grade 3/4 non-hematologic ADRs was ALT elevation (13.6%). PK analysis demonstrated that after multiple administrations the mean elimination half-life of clofarabine was 6.43 h, and the Tmax was around 2 h. The mean Cmax and exposure to clofarabine over the dosing period (AUC 0-24) achieved 581 ug/L and 2602.56 ug/L, respectively. It seems that there is no significant difference of PK profiles between Chinese and Caucasian patients.

Conclusion

Single clofarabine treatment is effective in Chinese pediatric patients with multiple relapsed or refractory ALL with acceptable safety profiles.

Disclosures:

No relevant conflicts of interest to declare.
Pegylated Asparaginase In Combination With Vincristine,Daunorubicin and Prednisone For Remission Induction In Adult Acute Lymphoblastic Leukaemia
Authors: Wei Na , Fu Li , Wang Jingshi , Wang Zhao
Published: October 2019

In contrast to that in children, the use of pegylated asparaginase in adult is very limited. We accept twenty-four newly diagnosed adult acute lymphoblastic leukemia (ALL) patients. Use pegylated asparaginase in combination with vincristine,daunorubicin and prednisone for remission induction. Pegylated asparaginase (Heng Rui Pharmaceuticals) administered at a dose of 2000 IU/m2 (maximum dose 3750 IU) intramuscular on day 10, 24. The aim was to observe complete response (CR) rate and side effect of the regimen. The overall CR rate was 91.67% (22/24), including 19 patients who were in CR after phase 1 of induction. 1 patient failed to achieve CR, and died of septicemia on day 24 of induction therapy. 1 Ph+ ALL patient got CR after Imatinib added. Grade 3 or 4 hematological adverse effects were seen in 100% patients. Most non-hematological adverse effects were grade 1or 2, including vomiting and nausea (87.5%), infection (66.7%), liver disfunction (50%), diarrhea (12.5) and pancreatitis (4.1%). No allergic reactions were observed. The level of fibrinogen and AT III descended in all patients, including 7 patients fibrinogen less than 1.0g/L, 14 patients AT III less than 50%. But no patient developed evidence of bleeding or thrombosis. This regimen got a satisfactory CR rate in adult ALL and was safe with few grade 3/4 side effects. Pegylated asparaginase was well tolerated, no allergic reactions evidence of bleeding or thrombosis was observed.

Disclosures:

No relevant conflicts of interest to declare.
Overcoming Drug Resistance Of FLT3-ITD By SAR302503
Authors: Kesarwani Meenu , Huber Erika , Azam Mohammad
Published: October 2019

A recent discovery of secondary mutations in the kinase domain of Fms-like tyrosine kinase 3 (FLT3) targeted by kinase inhibitor AC220 (quizartinib) established it as a relevant target in acute myeloid leukemia (AML). However, at the same time these observations suggest to identify next-generation inhibitors to target the resistant variants of FLT3. Here we show that SAR302503, a JAK2/FLT3 inhibitor in phase III trial for the treatment of polycythemia vera (PV) and myelofibrosis (MF), can potently inhibit FLT3-ITD variants that confer resistance to AC220. Interestingly, In vitro drug selection against SAR302503 at Cmax value (3 uM) failed to develop resistant clones suggesting it will be more effective in preventing the emergence of resistant clones. Structural modeling suggests that SAR302503 is a type I inhibitor that binds to ATP site in a DFG-in conformation. Altogether our data supports for clinical evaluation of SAR302503 for naïve and resistant variants of FLT3-ITD in AML patients to manage clinical resistance.

Disclosures:

No relevant conflicts of interest to declare.
Treatment With Leukapheresis In Patients Diagnosed With Hyperleukocytic Acute Myeloid Leukemia
Authors: Martínez-Cuadrón David , Montesinos Pau , Moscardo Federico , Lopez Lineth , Martin Guillermo , Solves Pilar , Boluda Blanca , Pérez-Sirvent Mariluz , Vera Belen , Navarro Ines , Alonso Carmen M , Martinez Jesus , López-Chulia Francisca , Sanz Jaime , Carpio Nelly , Rodríguez-Veiga Rebeca , Iacoboni Gloria , Sanz Guillermo , Sanz Miguel
Published: October 2019

Background

Patients with hyperleukocytic acute myeloid leukemia (AML) have a high mortality rate during induction treatment. This mortality is mainly due to bleeding complications, tumor lysis syndrome and symptoms of hyperleukocitosis. Although there is no clear evidence of benefit, leukapheresis is indicated to reduce the number of leukocytes (WBC) in peripheral blood and prevent or relieve leukostasis symptoms before starting cytotoxic therapy.

Objetives

Analyze the characteristics of a series of patients with hyperleukocytic AML who started induction treatment and assess the outcome depending on whether leukapheresis was performed or not.

Methods

All adult patients with hyperleukocytic AML (WBC> 95 x 109 / L) who received induction therapy in the Hospital La Fe between 1979 and 2012 were included. The leukapheresis was indicated according to physician discretion. The characteristics and mortality of both groups, patients who underwent leukapheresis and non-leukapheresis, were compared. Then a “matched-paired analysis” was performed according to ECOG, WBC and coagulopathy in a 2:1 ratio to compare results from two homogeneous cohorts.

Results

One hundred and forty patients diagnosed with hyperleukocytic AML received induction therapy, 18 of them underwent leukapheresis. Patients who underwent leukapheresis compared with non-leukapheresis showed a median age of 55 years (16-71) vs 58 (14-75) and WBC at diagnosis of 198 ´ 109 / L (101-620) vs 141 (97-375), respectively. The leukapheresis group had a worse ECOG (p = 0.03), more leukocytes (p = .003), more coagulopathy (p = .03), more leukostasis symptoms (p <.001) and more mortality at 7 days (p = .02). From the “matched-paired analysis”, 2 cohorts were obtained, whose characteristics are shown in Table 1. After comparing the group of leukapheresis and non-leukapheresis (n = 36), there were no significant differences between them, including leukocyte count, ECOG, coagulopathy and leukostasis. There were also no significant differences in mortality at 7, 14, 21 and 28 days (p = .60, p = .99, p = .92, p = .99, respectively).

Conclusions

Patients diagnosed with hyperleukocytic AML who underwent leukapheresis showed more leukocytes, leukostasis and coagulopathy. After analizying two comparable groups, leukapheresis failed to reduce mortality.

Disclosures:

No relevant conflicts of interest to declare.
STAT3 Predicts Poor Outcome In Patients With Advanced Diffuse Large B Cell Lymphoma
Authors: Feng Ru , Wei Xiaolei , Xu Meng , Huang Fen , Wei Yongqiang , Zhao Tong , Ye B. Hilda
Published: October 2019

Objective

To explore the expression and prognosis of STAT3 in Chinese patients with diffuse large B-cell lymphoma (DLBCL).

Methods

The expression of STAT3 was examined by the immunohistochemistry (IHC) method.120 patients with de novo DLBCL were reviewed retrospectively.

Results

Among the 120 patients, the ratio of male to female was 1.26:1, 47.5% of them were Non-GCB and 52.5% were GCB-DLBCL. The expression of STAT3 was observed in 50 of 120 cases (41.7%) and more frequent in the non-germinal center B cell-like (Non-GCB) DLBCL than that in the GCB subtype. The prognosis analysis showed that age, B symptom, bone marrow involvement, number of extranodal involvement, LDH, cell of origin and IPI related with prognosis, but not the expression of STAT3. However, in patients with advanced diffuse large B cell lymphoma, the expression of STAT3 was a poor prognostic factor.

Conclusion

STAT3 is expressed predominantly in Non-GCB DLBCL. The expression of STAT3 correlates poor outcome in patients with advanced diffuse large B cell lymphoma.

Disclosures:

No relevant conflicts of interest to declare.
A Therapeutic Vaccine In Adult T-Cell Leukemia Patients With Autologous Dendritic Cells Pulsed With Peptides Corresponding To Tax-Specific Cytotoxic T Lymphocyte Epitopes
Authors: Iino Tadafumi , Suehiro Youko , Hasegawa Atsuhiko , Watanabe Nobukazu , Tanosaki Ryuji , Utsunomiya Atae , Matsuoka Masao , Teshima Takanori , Akashi Koichi , Uike Naokuni , Okamura Jun , Kannagi Mari
Published: October 2019

Adult T-cell leukemia (ATL) is an aggressive peripheral T-cell neoplasm with extremely poor prognosis. It typically develops due to long-term infection with human T-cell leukemia virus type 1 (HTLV-1). Among HTLV-1 antigens, the viral regulatory protein Tax is widely recognized as a target of cytotoxic T lymphocytes (CTLs), and Tax-specific CTLs are often activated in ATL patients following hematopoietic stem cell transplantation (HSCT). This strongly suggests the presence of Tax expression in vivo and potential contributions of CTL to GVL effects. Based on these findings, we developed an anti-ATL therapeutic vaccine consisting of autologous dendritic cells (DC) pulsed with Tax peptides corresponding to the major epitopes of Tax-specific CTL that were previously identified from human leukocyte antigen (HLA)-A2, A24 or A11-possessing ATL patients post-HSCT (Cancer Res. 2004, J Virol. 2005). In preliminary experiments, the DC induced with a conventional method showed matured phenotype and produced interleukin (IL)-12 in two of three ATL patients tested.

After approval by the institutional ethics committees, we conducted a phase I clinical trial of anti-ATL immunotherapy for pretreated ATL patients in stable condition in order to evaluate whether HTLV-1 Tax targeting immunotherapy is safe and feasible. The vaccine protocol consists of three subcutaneous injections of peptide-pulsed DC at 2-week intervals in a total 8-week observation period to assess adverse events. Three acute-type patients enrolled and completed the course of the study. Modest clinical symptoms, such as injection site reaction, fever, and skin damage, were observed in all cases. DC vaccine-related toxicities were grade 1 or 2 and were considered relatively mild, and non-hematological toxicity was acceptable. In addition, significant reduction in sIL-2R levels were observed at least in one patient, and the HTLV-1 proviral loads were maintained below 60 copies /1000 PBMCs during the monitored period after administration of DC in the all patients. Enhanced proliferative responses of Tax-specific CTLs were also observed in vitro after therapy. Clinical effects, as evaluated by RECIST, were partial remission in two patients and stable disease in one patient. Durable remissions and improvements in performance status were observed in all three patients without the requirement for new treatment for 4-10 months after treatment with Tax-targeted DC vaccination. These results indicate that Tax-specific DC vaccination is safe and well tolerated, and may be promising as a new therapeutic strategy for ATL patients.

Disclosures:

No relevant conflicts of interest to declare.
Combination Of Arsenic and Interferon-a Inhibits Expression Of KSHV Latent Transcripts and Synergistically Improves Survival Of Mice With Primary Effusion Lymphomas
Authors: El Hajj Hiba , Ali Jihane , Ghantous Akram , Hodroj Dana , Daher Ahmad , Daher Ahmad , Zibara Kazem , Journo Chloé , Otrock Zaher K. , Zaatari Ghazi , Mahieux Renaud , El Sabban Marwan , Merhi Raghida Abou , Bazarbachi Ali
Published: October 2019

Background

Kaposi sarcoma-associated herpesvirus (KSHV) is the etiologic agent of primary effusion lymphomas (PEL). PEL cell lines infected with KSHV, but negative for Epstein-Barr virus have a tumorigenic potential in non-obese diabetic/severe combined immunodeficient mice and result in efficient engraftment and formation of malignant ascites with notable abdominal distension, consistent with the clinical manifestations of PEL in humans.

Methodology/Principal findings

Using this preclinical mouse model, we demonstrate that the combination of arsenic trioxide and interferon-alpha (IFN) inhibits proliferation, induces apoptosis and downregulates the latent viral transcripts LANA-1, v-FLIP and v-Cyc in PEL cells derived from malignant ascites. Furthermore, this combination decreases the peritoneal volume and synergistically increases survival of PEL mice.

Conclusion/Significance

These results provide a promising rationale for the therapeutic use of arsenic/IFN in PEL patients.

Disclosures:

No relevant conflicts of interest to declare.
ACase Of Asymptomatic Pediatric Chronic MyelogenousLeukemia With Hyperleukocytosis
Authors: Berger David A , Jaiyesimi Ishmael , Khandhar Paras
Published: October 2019

Introduction

Chronic Myeloid Leukemia (CML) represents 3% of pediatric leukemias. The natural history and molecular biology of pediatric CML is the same as those of older patients with CML. 95% of pediatric CML presents in asymptomatic chronic phase and the rest diagnosed in accelerated or in blast crisis stage. It is reported that patients with pediatric CML have higher white blood cell (WBC) counts and thereby have higher incidence of symptomatic leukostasis compared to adults. Sparse literature exists on particular triggers for treatment with leukapheresis (LA). We present a case of asymptomatic hyperleukocytosis (HL) secondary to pediatric CML and a brief discussion of treatment in this emergency.

Objective

To report a rare case of asymptomatic pediatric CML with HL.

Case Summary

A 9-year-old Caucasian male presented to his pediatrician on the outpatient basis for well child care and was found to have a WBC of > 200,000 and then was admitted to the Pediatric Intensive Care Unit for further management. Upon arrival in the PICU, pt was found to have splenomegaly (down 4-5 cm from the costal margin) and hepatomegaly (2 cm down from costal margin). CBC revealed hemoglobin of 7.4 g/dL, platelet of 883 bil/L and a total white blood cell count (WBC) of 302.7 bil/L. A differential on the WBC revealed 43.9 basophils, 24.2 blasts, and 6.1 promyelocytes. Flow cytometry showed prominent myeloid population with 5% CD34 positive myeloblasts and a prominent basophil population, consistent with morphological expression of CML. The cytogenetic analysis revealed a t(9,22)(q34;q11) translocation consistent with a Philadelphia chromosome. Pediatric Leukaphresis (LA) for cytoreduction was not available at our facility, thus he was transferred.

Discussion

The HL in this patient is a direct result of his pediatric CML. The decision regarding need for cytoreduction with LA has been described in the literature for symptomatic children, but no case has been reported of when to perform LA when no symptoms of leukostasis are present.

Conclusions

In this case, it was discovered that the trigger(s) to perform LA is vague in the literature, as well as controversial in practice especially with well appearing children with the potential for a swift precarious demise. Further studies and meta-analyses to uncover the proper triggers for LA are warranted.

Disclosures:

No relevant conflicts of interest to declare.
Application Of The Revised International Prognostic Scoring System In Korean Patients With MDS and Clinical Significance Of Clonal Evolution
Authors: Park Jae Hyeon , Park Si Nae , Kwon Jiseok , Im Kyongok , Kim Jung Ah , Choi Qute , Kim Seon Young , Hwang Sang Mee , Kim Inho , Yoon Sung-Soo , Park Seonyang , Lee Dong Soon
Published: October 2019

Introduction

The International Prognostic Scoring System (IPSS) was recently revised (IPSS-R). IPSS-R was developed using a large cohort of patients received supportive therapy, so we validated the new prognostic score system in Korean patients. IPSS-R emphasizes initial cytogenetic abnormalities, but the risk of clonal evolution is not well identified in follow up of MDS patients.

Methods

Data of 88 MDS patients were collected retrospectively and verified by chart review. And we also collected cytogenetic analysis results performed with bone marrow study.

Results

By IPSS-R cytogenetic scoring system, 2 (2.3%), 41 (46.6%), 22 (25.0%), 5 (5.7%), and 18 (20.5%) patients were classified as very good, good, intermediate, poor, and very poor, respectively. According IPSS, 5 (5.7%), 50 (56.8%), 19 (21.6%), and 14 (15.9%) patients were classified as low, intermediate-1, intermediate-2, and high, respectively. According to IPSS-R, 2 (2.3%), 13 (14.8%), 28 (31.8%), 25(28.4%), and 20 (22.7%) patients were reclassified as very low, low, intermediate, high, and very high, respectively. Median 3 years overall survival of patients intermediate, high and very high IPSS-R are 36 (95%CI 34-39), 32 (95%CI 8 – 56), and 13 (95%CI 6-21), respectively (P = 0.004). Clonal evolutions were occurred in 22 patients and they were classified by IPSS-R cytogenetic scoring system: 11 (50.0%) good, 8 (36.4%) intermediate, 1 (4.5%) poor, 2 (9.1%) very poor. Prior clonal evolution was considered as except concurrent occurrence of clonal evolution and acute myeloid leukemia (AML) transformation. These 17 patients have significant correlation with AML transformation (P = 0.010) and mortality (P = 0.021).

Conclusion

IPSS-R gives more refined prognostic discrimination and can be applicable in Korean MDS patients. In addition, the clonal evolution occurring during follow up should be considered as a risk factor of AML transformation and mortality.

Disclosures:

No relevant conflicts of interest to declare.
Utility Of Revised International Prognostic Scoring System (R-IPSS) To Predict Outcome Of Myelodysplastic Syndromes (MDS) Patients After Allogeneic Hematopoietic Stem Cell Transplantation (AHSCT)
Authors: Corrales-Yepez Maria Gabriela , Field Teresa , List Alan , Anasetti Claudio , Perkins Janelle , Al Ali Najla H , Mishra Asmita , Padron Eric , Lancet Jeffrey E , Kharfan-Dabaja Mohamed A. , Komrokji Rami S
Published: October 2019

Introduction

The R-IPSS at time of diagnosis was recently proposed and validated as a superior prognostic tool refining the original IPSS. The new model recognized further cytogenetic heterogeneity of the disease, and importance of the depth of cytopenia. The value of R-IPSS at time of AHSCT to predict post-transplant outcomes is not known. We examined the utility of R-IPSS prior to AHSCT in our cohort of MDS patients.

Methods

This was a retrospective study. We identified MDS patients who underwent AHSCT in our MDS database. Individual patient charts were reviewed. R-IPSS was calculated at time of transplant using data obtained from tests performed within 30 days of transplant. The R-IPSS karyotype was based on the new proposed 5 groups. The study main objective was to compare overall survival (OS) from time of AHSCT among R-IPSS groups and R-IPSS karyotype groups. Kaplan Meier estimates were used to calculate OS; log rank test was used to compare groups.

Results

Out of 1259 MDS patients in our database, 175 received AHSCT, and we excluded 65 patients who had AHSCT at any point after AML transformation. The median duration of follow up was 46 months (mo) (95% CI 39-53). Table-1 summarizes baseline characteristics of the cohort (n=110). All patients received a reduced intensity AHSCT using fludarabine and IV-busulfan. The rate of acute graft versus host disease (GVHD) II-IV was 68% and 72% for chronic GVHD. The median OS for the whole cohort was 22 mo (95% 10-33 mo.).

The IPSS at time of AHSCT did not predict OS. Based on R-IPSS, 24 pts (22%) were very low, 26 (24%), low, 16 (15%) intermediate, 21 (19%) high, and 23 (21%) were very high. The R-IPSS at time of AHSCT did not predict OS. The median OS was not reached, 14, 52, 20 and 23 mo for very low, low, int, high and very high respectively. (p= 0.1). The R-IPSS karyotype groups did predict OS. The median OS was 29 mo, 26 mo, 19 mo, and 9 mo for good, intermediate, poor and very poor karyotype risk groups respectively (p=0.047)

No other factors prior to AHSCT including age, myeloblasts percentage, Hemoglobin, platelets, treatment before AHSCT had impact on OS in univariate analysis.

Conclusions

IPSS and R-IPSS prior to AHSCT did not predict OS in our cohort of MDS patients. The R-IPSS karyotype group was the only statistically significant prognostic factor prior to AHSCT.

Disclosures:

List: Celgene: Membership on an entity’s Board of Directors or advisory committees.
Real-Life Efficacy Of Azacitidine In Myelodysplastic Syndromes According To IPSS Cytogenetic Profile
Authors: Breccia Massimo , Voso Maria Teresa , Maurillo Luca , Niscola Pasquale , Fianchi Luana , Aloe Spiriti Maria Antonietta , Buccisano Francesco , Pelliccia Sabrina , Fenu Susanna , Tendas Andrea , Latagliata Roberto , Alimena Giuliana
Published: October 2019

In the AZA001 trial, azacitidine prolonged OS in all cytogenetic IPSS risk subgroups and in particular reduced 67% risk of death for all patients with -7/del(7q). Aim of our study was to assess azacitidine efficacy according to cytogenetic risk at baseline in a large group of intermediate/high-risk MDS patients treated outside clinical trials. One hundred and sixty-six patients represented the whole cohort of patients with primary or secondary MDS (CMML were excluded) diagnosed and treated with azacitidine in 6 different hematologic units. Patients were recruited consecutively, without any criteria of exclusion. All patients received azacitidine with a schedule of 5+2+2 or of 7 consecutive days every 28 days until progression or unacceptable toxicity. Clinical parameters (age, sex, WHO classification, IPSS) and baseline cytogenetic evaluation were retrospectively collected. Of 166 patients recruited, 103 were males and 63 females; median age was 69.5 years (range 49-89). A median of 8 cycles was performed (range 1-60). According to IPSS stratification there were 29 patients with intermediate-1 risk, 118 patients with intermediate-2 and 15 high–risk (7 patients not determined). According to WHO classification, 37 patients were diagnosed as having RAEB-1, 101 patients as RAEB-2, 28 patients as RCMD. According to IPSS cytogenetic risk stratification, 88 patients were in the good risk, 22 patients were in intermediate risk and 39 in the poor risk. The most frequent cytogenetic aberration, apart from normal karyotype, was -7 or del(7q). According to cytogenetic stratification, after a median of 4 cycles for the first evaluation, we revealed the following responses according to IWG criteria: of 88 patients classified as good risk, 68 were evaluable and overall 30% achieved a complete (CR), partial remission (PR) or hematological improvement (HI), 59% maintained a stable disease, 4% progressed to acute leukemia and 7% failed to achieve any response. Of 22 patients classified as intermediate cytogenetic risk, 19 patients were evaluable after 4 cycles: overall, 52.6% achieved a CR/PR and 42% maintained a stable disease, none experienced a progression and 5% failed to achieve any response. Of 39 patients stratified as high cytogenetic risk, 30 were evaluable: 36.6% achieved a CR/PR/HI, 36% maintained a stable disease, 20% progressed to acute leukemia and 7% failed to achieve any response. Our results, based on a retrospective evaluation in a large series of patients treated outside clinical trials, shown that azacitidine was clinically effective independently from cytogenetic profile, as proved in the AZA001 trial.

Disclosures:

No relevant conflicts of interest to declare.
Effect Of Heterozygous JAK2 V617F On Thrombotic Risk In Patients With Polycythemia Vera: Measuring The Uncertain
Authors: Cacciola Emma , Cipolla Antonino , Di Francesco Ernesto , Seria Elisa , Torre Maria , Cacciola Rossella Rosari
Published: October 2019

The Janus Kinase (JAK)2 V617F mutation and its homozygous allele burden influences the thrombotic risk in Ph-negativemyeloid chronic disorders. Recent studies report that the vera polycythemia (PV) is a heterozygous JAK2V617F disease. The heterozygous PV is characterized by high hemoglobin (Hb), hematocrit (HCT) and red blood cell (RBC) and low thrombocytosis. Therefore, we evaluated allele burden, Hb, HCT, RBC and platelets and the occurrence of thrombotic events in patients with heterozygous PV disease. The study group included 50 patients (30 male, 20 female, mean age 68 years) affected by PV according WHO criteria. Their mean duration of disease was 11 years (range, 1-26 years). All patients were on aspirin. The percentage of granulocyte mutant alleles was evaluated using a quantitative real-time polymerase chain reaction-based allelic discrimination assay. Hb, HCT, RBC, platelets were measured by automated analyser. Of 50 patients, 28 had allele burden > 10% (34±5) and 22 had allele burden < 10% (6±1)). 10/28 had thrombosis (35%) including 6 transient ischemic attack (TIA), 2 myocardial infarction (MI) and 2 deep vein thrombosis (DVT), 18/22 (81%) had thrombosis including 8 transient ischemic attack (TIA), 4 myocardial infarct (MI) and 6 deep vein thrombosis (DVT). HB, HCT and RBC were elevated in all patients (19±1.8 g/dl, 56±4 %, 7±0.6 x106/L) and the platelets were higher in patients with allele burden > 10% than the patients with allele burden < 10% (500±127x109/L vs 381±128x109/L) (p = 0.030) whereas the thrombosis were more in patients with allele burden < 10% than the patients with allele burden > 10% (81% vs 35%). These data suggest that the heterozygous allele burden is a uncertain thrombotic risk factor.

Disclosures:

No relevant conflicts of interest to declare.
Mechanistic Insights Into Ventricular Arrhythmias Associated With Sickle Cell Disease: Role Of Abnormal Repolarization
Authors: Greener I D , Rutledge C A , Abassi T , Yadav K , Desai A B , Oberoi M , Chunduru P , Parikh D P , Garcia J GN , Gordeuk Victor R. , Hillery Cheryl A. , Dudley S c , Machado Roberto F , Desai A A
Published: October 2019

Introduction

Sudden death, ventricular arrhythmias, prolonged QTc on ECG, and myocardial fibrosis have been documented independently in patients with sickle cell disease (SCD). We hypothesized that these electrical abnormalities are associated with myocardial fibrosis, ventricular arrhythmias, and death in SCD.

Methods & Results

A retrospective chart review of 195 SCD patients seen at the University of Illinois at Chicago was performed evaluating the association of ECG intervals with documented mortality and/or ventricular tachycardia (VT). Only ECGs demonstrating normal sinus rhythm and heart rates < 100 were included. Significant increases in PR (181 + 30 vs 162 + 26 ms, p=0.009), QRS (102 + 24 vs 89 + 11ms, p=0.0002), QTc (456 + 36 vs 441 + 25, p=0.02), and Tp-Te (duration between peak to end of T wave in lead V5, 114 + 46 vs 76 + 23, p=2.11X10-7) intervals were associated with a combined endpoint of all-cause mortality and VT (n=16 compared to n=179 SCD controls) with similar heart rates across both groups (78 + 14 vs. 77 + 11, p=0.72). To elucidate the mechanisms underlying this increased clinical risk in SCD patients, we investigated hearts from a mouse model of SCD. Cardiac electrical stimulation in vivo induced ventricular arrhythmias in 3/4 homozygous (-/-) SCD compared to only 1/5 wild-type (WT) hearts. Interestingly, Tp-Te– an established sudden cardiac death predictor- was significantly prolonged in -/- SCD vs. WT mice (5.6 + 0.29 vs. 4.1 + 0.37 msec ; p<0.05). Furthermore, left ventricular (LV) effective refractory periods (37+ 2.3 vs. 22 + 4.57 ms; p<0.05) and mid-anterior (LV) monophasic action potentials (MAPs) of -/- SCD mice, revealed increased duration compared to WT mice (53 + 2.3 vs. 33 + 6.9 ms; p<0.05).

Conclusion

In SCD patients, poor clinical outcomes are associated with abnormalities in depolarization and, prominently, repolarization. The SCD mouse exhibited arrhythmia vulnerability associated with repolarization abnormalities (Tp-Te, LV MAPs). The SCD mouse may represent a useful model for deciphering the mechanisms underlying the apparent increased arrhythmia burden in SCD patients.

Disclosures:

Hillery: Bayer: Consultancy; Biogen Idec: Consultancy.
Genetics and Epigenetics of Fetal Hemoglobin Control
Authors: Orkin Stuart H.
Published: October 2019

Expression of fetal hemoglobin (HbF, α2γ2) greatly ameliorates the severity of the major hemoglobin disorders, sickle cell disease (SCD), and the β-thalassemias. Efforts to reactivate HbF in adults with these disorders have relied on empirical observations or therapeutic modalities that are indirect. A major goal for the field is the development of targeted reactivation of HbF through relief of γ-globin gene silencing. The regulatory factors that participate in the switch from HbF to HbA in ontogeny and in γ-gene silencing in the adult have been elusive, therefore precluding mechanism-based reactivation of HbF. Recent findings, largely derived from genome-wide association studies (GWAS), have transformed the current understanding of globin switching. This presentation will review recent evidence supporting direct involvement of the zinc-finger repressor protein BCL11A in both developmental switching of globins and HbF silencing in the adult. These studies include the impact of naturally occurring genetic variation at the BCL11A locus on HbF levels, proof-of-principle experiments in genetically engineered mice suggesting that interference with BCL11A action alone may be sufficient to provide therapeutic elevation of HbF, and the nature of protein partners of BCL11A that may mediate some aspects of BCL11A function. Recent findings on the manner in which genetic variation within the BCL11A locus influences BCL11A expression provide special insight into quantitative aspects of HbF regulation and raise the possibility of new strategies to cripple BCL11A. The opportunities and challenges for the development of mechanism-based reactivation of HbF will be discussed in the context of ongoing efforts to exploit small molecule and genetic approaches. The tools are in hand to translate an improved understanding of globin gene regulation for the benefit of patients with the major hemoglobin disorders.

Disclosures:

No relevant conflicts of interest to declare.