bluuebunny/crossref_metadata_2025 huggingface dataset:
Disproportionate Sex Chromosome Loss in Abnormal Karyotypes of Male and Female Patients with Diagnoses of Myeloid or Lymphoid Neoplasms
Authors: Cantu Eduardo S. , Nemana Lakshmi , Pierre Robert V. , Moses Mike
Published: October 2019

Sex chromosome loss (SCL) occurs in different types of human cells and is likely related to aging, but could also be related to other factors. We reviewed cytogenetic studies of bone marrow samples from patients (males and females) with clinical diagnoses of either myeloid or lymphoid delineation in order to establish if SCL is associated with abnormal karyotypes found in these malignancies. Prevalence of clonal SCL (POC-SCL = % cases having ≥ 3 cells with SCL/study) was evaluated retrospectively in a total of 9,593 bone marrow cytogenetic studies performed in our laboratory during a recent calendar year. Results from this database were identified as normal or abnormal karyotypes, as male or female patients, and as having a myeloid or a lymphoid diagnosis. POC-SCL values were determined for each of these designations. Student-t tests were then applied to ascertain if POC-SCL values were significantly different with respect to these three variables. Our statistical analyses showed that: 1) POC-SCL was higher in abnormal than in normal karyotypes. This was true for abnormal male and female karyotypes in both myeloid and lymphoid disorders, compared to cases with normal karyotypes. 2) POC-SCL values were higher in male karyotypes for normal and abnormal myeloid cases, than in females. This was also true in normal karyotypes of lymphoid cases, but had similarly high values for both genders in abnormal karyotypes of lymphoid cases. 3) POC-SCL was about the same in total myeloid versus total lymphoid cases. However, closer examination indicated a higher POC-SCL value was seen in lymphoid patients with normal male and female karyotypes than for myeloid patients with normal male and female karyotypes. Interestingly, POC-SCL was higher in the abnormal myeloid male karyotypes than in the normal male karyotypes, while it was higher in abnormal lymphoid female karyotypes than in the normal female karyotypes. Although clonal-SCL may not reflect a primary biological marker associated with these cancers, our results clearly show that clonal-SCL segregates selectively with abnormal karyotypes associated with myeloid and lymphoid malignancies. Moreover, not only is there a disproportionate over-representation of -Y in abnormal karyotypes, but –X is also a significantly frequent finding in abnormal karyotypes of these diseases.

Disclosures

Cantu: Integrated Oncology: Employment. Nemana:Integrated Oncology: Employment. Pierre:Integrated Oncology: Consultancy. Moses:Integrated Oncology: Employment.
OCT1 gene Expression Is an Independent Prognostic Factor in Diffuse Large B Cell Lymphoma
Authors: Gouveia Gisele Rodrigues , Ferreira Suzete Cleusa , Siqueira Sheila , Pereira Juliana
Published: October 2019

Background: Transcription factors associated with the POU domain modulate the expression of several genes of B lymphoid differentiation, including the IgH. The study of these factors allowed to better understand the pathogenesis of lymphomas and to establish the lineage and the differentiation stage of the malignant cell. The silencing of OCT1 in tumor cell lines reduced its malignant transformation, but its ectopic expression enhanced the tumorigenesis ability. However, few studies has been evaluated the role of the OCT1 gene expression in lymphoma. In this study we assessed the impact of the OCT1 gene expression in the survival of patients with Diffuse Large B Cell Lymphoma (DLBCL).

Methods: From January 2006 to January 2011 were evaluated 77 patients with DLBCL treated with R-CHOP at Clinical Hospital and Cancer Institute of Sao Paulo University. The RNA was extracted from the paraffin block at lymphoma diagnosis and gene expression analysis was performed by relative quantification method by Real-Time PCR (qRT-PCR). After the data normalization using two different reference genes, the median expression of OCT1 was obtained. The overall survival (OS) and progression-free survival (PFS) were estimated by the Kaplan-Meier method. The relative risks were obtained by Cox regression bivariate intervals with 95% of confidence. The significance level of 5% was accepted and the IBM SPSS Statistics software version 20.0 was used.

Results: Patients showing OCT1 expression < the median presented higher OS (p = 0.010) and PFS (p = 0.016) than patients with OCT1 expression ≥ median with a hazard rate (HR) for OS and PFS of 2.45 and 1.14, respectively. In multivariate analysis the PFS was also higher in patients with OCT1 expression < the median (p = 0.035). The stratification by the international prognostic index (IPI) and age showed that the expression of OCT1 < median showed a statistically significant difference in the OS (p = 0.048) in IPI intermediate-high (HI) and high (HR) patients (p = 0.048), with a HR of 2.32 in HI plus HR group. The PFS (p = 0.025) and OS (p = 0.025) were lower in patients ≥ 60 years and OCT1 expression ≥ the median.

Conclusion: Our data suggest that the expression of OCT1 showed a predictive prognostic impact in DLBCL independently of IPI. Patients with lower expression of OCT1 presented a better OS and PFS.

Figure 1 SG curve for the OCT1 gene expression. Figure 1. SG curve for the OCT1 gene expression.

Figure 2 SLP curve for the OCT1 gene expression. Figure 2. SLP curve for the OCT1 gene expression.

Figure 3 SG curve for to expression of OCT1 gene for subgroup IPI intermediate-high and high. Figure 3. SG curve for to expression of OCT1 gene for subgroup IPI intermediate-high and high.

Disclosures

No relevant conflicts of interest to declare.
Gambogic Acid Suppresses Hypoxia-Induced HIF-1α/VEGF Expression Via Inhibiting PI3K/Akt/mTOR Pathway in Multiple Myeloma Cells
Authors: Wang Fei , Zhang Wei , Guo Liting , Cheng Jian , Liu Ping , Chen Baoan
Published: October 2019

In multiple myeloma (MM), the hypoxic environment is an important factor causing tumor angiogenesis which is strongly correlated to disease progression and unfavorable outcome by activating the key transcription factor, hypoxia-inducible factor-1α (HIF-1α). Gambogic acid (GA) is the major active ingredient of gamboge, which has been shown to possess antitumor effect by in vitro and in vivo study. However, the underlying molecular mechanism that whether GA inhibits tumor angiogenesis remains not well understood. In this study, we investigated the effects of GA on expression of HIF-1α, and its downstream target gene vascular endothelial growth factor (VEGF) in human MM U266 cells. We found that hypoxia induced increase in the level of HIF-1α subunit protein and activated the PI3K/Akt/mTOR pathway. Moreover, the treatment with GA markedly decreased HIF-1α and VEGF expression under hypoxic condition. Mechanistic studies exhibited that GA inhibited the production of HIF-1α by reducing phosphorylation of Akt and mTOR in U266 cells. Furthermore, in vivo study revealed that intravenous injection of GA once every other day for two weeks could suppress tumor volumes by antiangiogenesis activity. Taken together, our results identify GA suppress hypoxia-activated pathways that linked to MM progression, at least partly, by the inhibition of PI3K/Akt/mTOR signaling pathway. Therefore, GA may be a new potent therapeutic agent against human MM cells.

Disclosures

No relevant conflicts of interest to declare.
Effect of DMSA Modified Fe3O4 and Multidrug Resistance Modulator HZ08 on the Reversal of Multidrug Resistance in K562/A02 Cell Line
Authors: Shen Fei , Zhou Mingfei , Lu Xuzhang , Chen Yanping , Chen Baoan
Published: October 2019

The objective of the present study was to investigate the reversible effect of HZ08 and daunorubicin(DNR) combined with dimercaptosuccinic acid modified iron oxide (DMSA-Fe3O4) magnetic nanoparticles (MNPs) in human chronic leukemia cell line K562/A02, and the mechanism potentially involved. The growth inhibition rate of K562/A02 cells was determined by Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis and intracellular concentration of DNR were detected by flow cytometry (FCM). DAPI staining was used to view apoptotic cellular morphology. Subsequently, transcription levels of MDR1 mRNA and expression levels of P-glycoprotein (P-gp) and caspase-3 were determined by real time polymerase chain reaction (real-time PCR) and Western blotting analysis, respectively. group clearly exhibited more morphological changes (severe structural alterations) than other groups. In addition, transcription of MDR1 gene and protein expression of P-gp and caspase-3 of K562/A02 cells were regulated at the most remarkable extent in DNR-HZ08-MNPs group when compared with other groups. These findings suggest that the remarkable effects of the novel DNR-HZ08-MNPs on multidrug resistant K562/A02 leukemia cells would be a promising strategy for overcoming multidrug resistance.

Disclosures

No relevant conflicts of interest to declare.
Small Molecule ONC201/TIC10 Induces Caspase-Dependent Apoptosis in Acute Lymphoblastic Leukemia Cells Via Modulation of Bcl-2 and IAP Family Proteins
Authors: Prabhu Varun Vijay , Lulla Amriti R. , Wagner Jessica M , Hernandez-Borrero Liz J. , Talekar Mala K. , Kline Christina L. , Dicker David T. , Barth Brian M , Pu Jeffrey J. , Claxton David F. , Allen Joshua E. , El-Deiry Wafik S.
Published: October 2019

ONC201/TIC10 is a potent small molecule anti-tumor agent in several types of solid tumors and lymphomas. ONC201/TIC10 is on track to enter clinical trials for patients with advanced cancer in 2014, with IND issued by the FDA in March, 2014. Early trials will evaluate the safety and efficacy of ONC201/TIC10 as a monoagent in hematological malignancies. In the current study, we evaluated the anti-cancer effects of the small molecule in Acute Lymphoblastic Leukemia (ALL). Analysis of cell viability by the CellTiter-Glo method revealed that ONC201/TIC10 treatment reduces the viability of three ALL cell lines (Reh, Jurkat, MOLT-4) in a dose- (2.5/5/10 μM) and time-dependent manner (24/48/72 h). We have previously reported that ONC201/TIC10-mediated reduction in cell viability and apoptosis in various types of solid tumors occurs at 60/72 h. Interestingly, ONC201/TIC10 reduces the viability of ALL cell lines within 24/48 h at the indicated doses. An inactive TIC10 isomer compound synthesized by Medkoo Biosciences with a structure related to the active ONC201/TIC10 compound does not reduce the viability of ALL cells. Sub-G1 analysis indicated that ONC201/TIC10 induces apoptosis in ALL cells and a pan-caspase inhibitor reduces ONC201/TIC10-mediated apoptosis. Western blot analysis was used to further investigate the mechanism of ONC201/TIC10-mediated apoptosis. ONC201/TIC10-mediated apoptosis involves PARP cleavage and caspase-9 activation. Anti-apoptotic Bcl-2 family members Bcl-2 and Bcl-xl are downregulated while the pro-apoptotic Bcl-2 family member Bim is upregulated in response to ONC201/TIC10 treatment. ONC201/TIC10 also downregulates the inhibitor of apoptosis (IAP) family proteins cIAP1 and cIAP2. We have previously shown that the anti-tumor effect of ONC201/TIC10 involves inhibition Akt and ERK phosphorylation resulting in Foxo3a activation and TRAIL-gene transcription. We observed inhibition of Akt phosphorylation upon ONC201/TIC10 treatment of ALL cells. Thus, ONC201/TIC10 holds promise as a novel agent for the treatment of ALL based on its robust activity in preclinical models of the disease. Our ongoing studies are evaluating the impact of this novel therapy on ALL cells with different translocations, and are introducing combination therapy with ONC201/TIC10 for ALL.

Figure 1 Figure 1.

Disclosures

Allen: Oncoceutics: Employment, Equity Ownership, Patents & Royalties. El-Deiry:Oncoceutics, Inc.: Equity Ownership, Patents & Royalties.
The Outcome of Childhood Acute Lymphoblastic Leukemia with MLL Gene Rearrangement Treated with the Protocol (CCLG-ALL2008)
Authors: SUN Yina , Chai Yi-huan , He Hai-long , LU Jun , WANG Yi , Zhao Wen-li , Xiao Pei-fang , Fan Jun-jie , Hu Shaoyan
Published: October 2019

Objective: MLL gene rearrangement in pediatric acute lymphoblastic leukemia (ALL) was regarded as high risk factor because of the poorer outcome of overall survival(OS). Some reports further proved that patients with MLL/AF4 had the worse outcome than MLL rearranged with other partners. CCLG-ALL2008 protocol has been carried out in China for more than 5 years. However, there was no reports to evaluate its efficiency on pediatric ALL patients with MLL rearrangement. In this study, we analyzed the data of ALL patients to compare the outcome of patients with MLL rearrangement positive and negative treated by CCLG-2008 protocol.

Methods During the period from 2009 to 2013, 379 patients were enrolled in this protocol, of which 19 cases were MLL rearrangement positive and treated with CCLG-ALL2008 protocol for high-risk (HR) group. The treatment efficiency was evaluated on the time points of day 7, day15, day 33 and 12th week after treatment, respectively. OS and treatment-related mortality (TRD) was calculated within high risk groups of MLL positive and negative.

Results Patients with MLL rearrangement positive accounted for 5.01% of all patients. The characteristics and response to the treatment were illustrated in Table 1. Cases younger than 2 years old, with initial white blood cell (WBC) 50*109/L , or MRD more than 10-2 on day 33 had a lower OS (P<0.05). Compared with high risk group with MLL negative, MLL rearrangement positive group didn't show any statistic difference in the recurrence rate, TRD, and 5 years event-free survival (EFS)(P>0.05). Compared with the male, female patients had a better 5 year EFS (100% versus 50%±17%, P<0.05) (Figure 1). Age did matter in MLL rearrangement positive group, older than 2 years cases had a better 5 years EFS than those younger than 2 years old (86%±11% versus 27%±11%, P<0.05) (Figure 2). Multivariate COX regression analysis found that prednisone response on day 7 has impacted a role on OS (P<0.05). Other factors such as gender, age (older than 2 years), WBC count, MLL fusion partners, and MRD on day 15 and day 33 and week 12th didn't show statistic influence on OS in ALL patients with MLL gene rearrangement treated by the protocol of CCLG-ALL2008.

Conclusion Factors including female, older than 2 years of age, sensitivity to prednisone, are good prognostic index in predicting the outcome of 5 years of OS and EFS in childhood ALL with MLL gene rearrangement.

Table 1 Clinical and laboratory characteristics of ALL patients with MLL positive Table 1. Clinical and laboratory characteristics of ALL patients with MLL positive

Figure 1 Gender exerts influence on 5 years EFS in MLL rearrangement group Figure 1. Gender exerts influence on 5 years EFS in MLL rearrangement group

Figure 2 Figure 2. Age influenced 5 years EFS in MLL rearrangement group

Disclosures

No relevant conflicts of interest to declare.
Low-Dose Arsenic Trioxide Combined with Aclacinomycin a Synergistically Enhance the Cytotoxic Effect on Human Acute Myelogenous Leukemia KG-1a Cell Line By the Induction of Apoptosis
Authors: Ye Yongbin , Xu Xiaojun , Zhang Mingwan , Qiu Dafa , Bai Xiaochun , Wang Jing , Weng Guangyang , Ruiqing Zhou , Guo Ziwen , He Huiqing , Yi Wenfang , He Xin , Kunyuan Guo
Published: October 2019

Acute myeloid leukemia (AML) is a kind of common disorder in the elderly. Although remarkable progress has been made in the treatment of this deadly disease over the past few decades, the outcome remains poor. While conventional chemotherapy may be effective for some elderly patients, the vast majority do not tolerate intensive conventional treatment well, thus the development of a more effective method to overcome this problem is necessary. Aclacinomycin A (ACM), an antitumor antibiotic, was used to treat various tumors including AML. However, its high toxicity limits its clinical application. Previous studies reported that arsenic trioxide (As2O3) has shown substantial efficacy in treating patients suffering from AML. However, the molecular mechanisms of As2O3-induced cytotoxic effect of AML cells are not yet fully known. The aim of this study was to investigate the synergistic cytotoxic effect of low-dose As2O3 combined with ACM on human KG-1a cells and to clarify the possible mechanism underlying this combination treatment. Our results showed that As2O3 or ACM inhibited proliferation of KG-1a cells in a time- and dose-independent manner. In the flow cytometric analysis, the low-dose of the matched combination treatment induced a more prominent apoptosis than the single agent-treated groups. Furthermore, western blot analysis showed that the combination treatment decreased Bcl-2 expression and increased caspase-3 expression more significantly than the single drug treatments. More importantly, we showed that the combination index (CI) values were < 1 in all matched combination groups analyzed by compusyn software. In conclusion, our data showed that low-dose As2O3 combined with ACM showed a more prominent cytotoxic effect than single agent treatment on KG-1a cells. These results may provide a high efficiency but low-toxicity therapeutic benefits for AML.

Disclosures

No relevant conflicts of interest to declare.
High Expression of Mir-15a Predicts Shorter Survival and Worse Response to Chemotherapy in Patients with Acute Myeloid Leukemia (AML)
Authors: Butrym Aleksandra , Baczynska Dagmara , Tukiendorf Andrzej , Rybka Justyna , Dobosz Tadeusz , Jurczak Wojciech , Kuliczkowski Kazimierz , Mazur Grzegorz
Published: October 2019

Background: MicroRNAs (miRNAs) are small non-coding RNA molecules, that control gene expression by targeting messenger RNA (mRNA), via degradation or suppression of translation. Aberrant expression of microRNAs (miRs) has been proved to have a role in acute myeloid leukemias (AML)

The aim of the study was to determine expression of miR-15a in acute myeloid leukemia patients before and after chemotherapy and its influence on patient clinical outcome.

Methods: miRNAs from isolated leukemic cells were extracted using mirVanaTM miRNA Isolation kit (Ambion Inc., Carlsbad, CA, USA) following the manufacturer's protocol. Reverse transcriptase (RT) reactions were performed for mature miRNA cDNA synthesis in separate tubes using specific stem-loop RT primers and TaqMan® MicroRNATM Reverse Transcription kit (Applied Biosystems, Foster City, CA, USA). After microRNA isolation, reverse transcriptase reactions were performed, followed by cDNA amplification. The relative amount of microRNA-15a was normalized according to the reference RNU48 level. Results were considered statistically significant with p-value < 0.05.

Results: 95 patients (aged 60.2 ± 15.0, 22–90, Male = 61%) with newly diagnosed AML were included into the study. Samples of the bone marrow for miR-15a expression analysis were collected before start of chemotherapy and repeated after completed induction chemotherapy (40 patients). A control group of 20 matched patients was also taken into account. The analyzed group consisted of: 7 patients with AML M0, 34 had M1, 29 had M2, 14 had M4 and 11 had M5.

AML patients has higher miR-15a expression than control group (p=0.005633). The risk of death in AML patients was higher in patients with higher miR-15a expression on diagnosis (p=0.0430), Fig.1. Patients with lower miR-15a expression were more likely to achieve complete remission after induction chemotherapy (p=0.0130). After successful chemotherapy we observed significant drop in miR-15a expression.

Conclusions: We proved that miR-15a was upregulated in AML patients and its expression influenced remission achieving and death risk.

Figure 1. Survival of AML patients depending on miR-15a expression. Figure 1. Survival of AML patients depending on miR-15a expression.

Disclosures

No relevant conflicts of interest to declare.
Hodgkin’s Disease (1975-2012): Long-Term Results of a Single Center
Authors: Celkan Tiraje , Özdemir Gül Nihal , Akı Hilal , Apak Hilmi , Özkan Alp , Hallaç Metin , Adaletli İbrahim , Özcan Rahşan , Ergen Şefika Arzu , Yıldız İnci
Published: October 2019

Aim: To investigate the clinical features, diagnostic and treatment results and long-survival of children with HL at a single center in Turkey.

Methods: Retrospective data were extracted from medical records of children (<18 years old) diagnosed and treated for HL between 1975 and 2012 at Istanbul University, Cerrahpasa Medical Faculty.

Results: Twohundred-fortyeight patients (M/F: 1.8) were identified for analysis. Mean age at diagnosis was 10 years. Four-percent were stage 1, 43% were stage 2, 34% were stage 3 and 17% were stage 4 at diagnosis. B symptoms were present in 105 children (42%). Mixed cellular was the most common subtype (50%) followed by nodular sclerosis. MOPP regimen was primarily used until 1989 along with extended field radiotherapy (30-40 Gy) after 1989 ABVD with involved field RT (20-25 Gy) was the treatment of choice. Onehundred-fortyseven patients (42%) had radiotherapy. Median follow-up time was 14.3 years (range: 5 months-38 years). PET was used for assessment in 45 cases. Ten year overall survival rate was 99%. Ten year event free survival rate (EFS) was 90%. Univariate analysis showed that stage, histological subtype, presence of B symptoms and hematological parameters were not related to OS while treatment regimen correlated. Twenty-six patients relapsed and 17 patients expired on follow-up. Ten patients who relapsed were treated with autologous stem cell transplantation, 3 of them relapsed again and were treated with brentixumab. Secondary tumours developed in 4 patients (1.6%). One had AML, 1 osteosarcoma, 1 thyroid papillary cancer and 1 had craniopharyngioma.

Conclusion: Ethnic and regional differences in the epidemiology and pathological aspects of HL may be associated with differences in clinical features and prognosis however we found that clinical features and treatment outcomes are similiar in Turkish patients. Our results showed that hematological parameters and B symptoms were not significant prognostic parameters.

Disclosures

No relevant conflicts of interest to declare.
Risk of Second Malignancies after Non Hodgkin Lymphoma: A Cohort Study of 22,466 Survivors in Israel
Authors: Tadmor Tamar , Barbara Silverman , Liphshiz Irena , Polliack Aaron
Published: October 2019

Previous studies have shown an increase risk of second malignancies after non-Hodgkin's lymphoma (NHL). This is probably related to genetic predisposition, molecular and pathologic background, immunological status and type of therapy given for the lymphoma.

We evaluated the risk of second solid tumor and hematological malignancy among 24,666 survivors of NHL diagnosed in Israel during 1980-2011. Data was taken from the records of the Israeli National Cancer Registry (INCR), as all public and private medical facilities, and pathology laboratories engaged in diagnosing cancer patients in Israel, are obliged to send a copy of the medical summary, demographic data and details of diagnosis, to the registry. The INCR records all the available information on these cases, including the date of diagnosis and tumor location.

The cohort of 24,666 NHL-patients included Jews (22,601 cases) and Arabs (2065 cases). Median age at diagnosis of Jews was 61.3 years, and 48.2 for Arab patients. Of all Jews with NHL: 11,265 (50%) were of European-American descent, 5005 (22%) Asian African and 6114 (27%) were born in Israel. In both Jewish and Arabs sub-groups, there were more males (52%, 56% respectively). Second cancers were reported in 2010 patients; 1918 Jews (95.5 %), and 92 Arabs (4.5 %). Evaluation of the results was restricted to the cohort of Jews only, due to the small number of Arab patients.

Second malignancies in all recorded sites were significantly more common in males with standardized incidence ratio (SIR) of 1.28, and 1.25 for females. This was even more pronounced for secondary hematological malignancies, with SIR of 1.97 and 1.81 respectively, in a total of 283 cases. (Table)

Of the secondary hematological malignancies; the most frequent were leukemias (31%) and NHL (29%), followed by multiple myeloma (14%), Hodgkin lymphoma and mycosis fungoides (both 8%).

In conclusion; there is an increased risk of second malignancies after NHL in Israel, particularly leukemia and non- Hodgkin lymphoma. These clearly occur more frequently in Jews, suggesting that ethnicity and genetic susceptibility may be important risk factors. TableSecond cancer after NHL in Jews ( 1980 – 2011)All sitesObservedExpectedSIR 95% CImale1056822.181.281.211.36female862689.861.251.171.33HematologyObservedExpectedSIR 95% CImale16483.401.971.672.27female11965.611.811.492.14

Disclosures

No relevant conflicts of interest to declare.
The Expression of CD44v6 Predicts Poor Outcome in Patients with Diffuse Large B Cell Lymphoma Treated with R-CHOP
Authors: Wei Yongqiang , Xie Muchen , Huang Fen , Wei Xiaolei , Jing Hui , Hao Xiaoxiao , Zeng Hong , Zhou Qinjun , Xu Linwei , Feng Ru
Published: October 2019

Background:

CD44 variants have been implicated in metastasis and as an unfavorable prognosis factor in many types of cancers. Our previous study had showed that CD44v6 expression implied a poor clinical outcome in diffuse large B cell lymphoma (DLBCL) patients treated with CHOP. However, it is remains unknown the prognostic value in DLBCL patients treated with R-CHOP due to our sample sizes. Therefore, to examine the prognostic value of CD44v6 expression in DLBCL patients, we performed this retrospective study.

Methods: All 141 patients diagnosed as de novo DLBCL according to WHO classification were further confirmed. All patients treated with R-CHOP (rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone). The expression of CD44v6 was analyzed by immunohistochemistry (IHC). IHC was performed by an automated immunostainer complying with the instructions of the protocols.

Results: Among all the 141 patients, the expression of CD44v6 was observed in 41 cases (29.1%). CD44v6 is expressed predominantly in non-germinal center type DLBCL. There was no significant difference in gender, age, B symptoms, performance status, LDH, stage and IPI score between patients with and without CD44v6 expression. Patients with CD44v6 expression showed significant inferior overall survival, but not event-free survival compared with CD44v6-negative patients. (p=0.022 and p=0.142, respectively).Multivariate analysis showed that the expression of CD44v6, independent of the international prognostic index and cell of origin, implied a poor overall survival (HR=2.223; 95% CI= 1.007-4.906, p=0.048), but not event-free survival (HR=1.457; 95% CI=0.773-2.745, p=0.245).

Conclusions: These data suggest that the expression of CD44v6 implied poor outcome in DLBCL patients treated with R-CHOP.

Disclosures

No relevant conflicts of interest to declare.
The Hybrid Regimen of ABVD/BEACOPP May Overcome the High-Risk Features of the Advanced Stage Hodgkin Lymphoma
Authors: Alaskar Ahmed , Al Ameer Ahmad , Ahmed Gasmelseed , Alhejazi Ayman , Abuelgasim Khadega , Gmati Giamal , Rather Mushtaq , Al Zahrani Mohsen
Published: October 2019

Background: Hodgkin Lymphoma (HL) has improved markedly overtime. This study aims to assess our experience in treating advanced HL.

Method: In a retrospective cohort study, patients age >14 with advanced HL from 2007-2010 were included. Using IPI, patients were categorized into low and high risk. ABVD and Hybrid ABVD+BEACOPP regimens were used.

Result: 74 patients were included with the median age at diagnosis of 25.5 years. 50% presented with stage IV. Two different protocols were used (ABVD and Hybrid ABVD + BEACOPP). Median follow up was 30.3 months. Male gender and stage V were most often to receive hybrid regimen (odd ratio 3.74 and 6.58 respectively). When the two groups were compared, the higher risk group (mostly treated with hybrid regimen) showed similar survival rates to the lower risk group (treated mostly with ABVD) with p-values of 0.41 for OS and 0.42 for PFS.

Conclusion: This study shows quite similar results to the published studies in the OS and PFS with a tendency to overcome the high-risk features by the hybrid regimen in advanced HL. However, this needs to be confirmed in a prospective randomized study.

Disclosures

No relevant conflicts of interest to declare.
Haploidentical Hematopoietic Stem Cell Transplantation for Acquired Severe Aplastic Anemia
Authors: Miao Miao , Zhang Xiang , Xu Ting , Jin Song , Wang Hong , Yang Xiaofei , Pan Zhijuan , He Jun , Sun Aining , Depei Wu
Published: October 2019

Objective: To evaluate the efficacy and safety of haploidentical hematopoietic stem cell transplantation (haplo-HSCT) in patients with acquired severe aplastic anemia (SAA), who lacked suitable related or unrelated HLA-matched donors.

Methods: 39 SAA patients underwent haplo-HSCT from Jul 2012 to Jun 2015 at our center. There were 23 males and 16 females at a median follow-up of 11 (range, from 0 to 36) months. The median time from diagnosis to transplantation was 1 (range, from 0.5 to 52) months. The median ages of SAA patients and related haploidentical donor were 23 years (range, 9 to 51years) and 45 (range, from 21 to 61) years, respectively. All patients were given BuCy plus ATG conditioning regimen. GVHD prophylaxis regimen consisted of cyclosporine A (CsA), mycophenolate motetil (MMF), and short-term methotrexate.

Results: Stem cells were collected from bone marrow in 23.08% (n=9) of patients, peripheral blood in 2.56% (n=1), bone marrow plus peripheral blood in 74.36% (n=29) patients. 36 patients received haplo-HSCT combined with the third part of cord blood transfusion 92.31%. The median stem cell dose transplanted was 9.76 (range, from 4.02 to 20.10)×108/kg for mononuclear cells, while 3.4 (range, from 1.05 to 8.60)×108/kg for CD34 cells. 36 patients achieved neutrophil engraftment at a median of 12 (range, from 9 to 28) , and 29 patients achieved platelet engraftment at a median of 29 (range, from 10 to 26) days. Cumulative incidence of III°~IV° acute graft versus host disease (aGVHD) was 8.9±4.9%. 6 patients died of transplant-related mortality (TRM), including 4 from severe infection, and 1 from TMA. The overal survival rate of all patients was 83.2%±6.4%

Conclusions: Haplo-HSCT is likely to be an option for SAA patients without suitable related or unrelated HLA-matched donors, in consideration of the acceptable TRM and severe GVHD incidences.

Disclosures

No relevant conflicts of interest to declare.
The RIG-I Agonist 3pRNA Synergizes with Checkpoint Blockade in Cancer Immunotherapy
Authors: Heidegger Simon , Kreppel Diana , Bscheider Michael , Wintges Alexander , Bek Sarah , Schmickl Martina , Ruland Jürgen , Peschel Christian , van den Brink Marcel , Haas Tobias , Poeck Hendrik
Published: October 2019

Antibody-mediated targeting of regulatory T cell receptors such as CTLA-4 has been shown to enhance anti-tumor immune responses against several cancer entities including malignant melanoma. Yet, therapeutic success in patients remains variable underscoring the need for novel combinatorial approaches. Here we established a vaccination protocol that combines selective engagement of the nucleic acid-sensing pattern recognition receptor RIG-I, antigen and CTLA-4-blockade. We found that vaccination together with RIG-I ligation strongly synergized with CTLA-4 blockade to induce expansion and activation of antigen-specific CD8+ T cells and potent anti-tumor immunity. Cross-priming of cytotoxic T cells as well as anti-tumor immunity required the adapter protein MAVS and type I interferon (IFN) signaling and were mediated by dendritic cells. In addition, the benefit of the combined immunization with anti-CTLA-4 was reduced by systemic antibiotics pointing to the requisite of an intact commensal microbiota in this context. Together, our findings describe a novel combinatorial strategy that may form the basis for the design of new type I IFN-based regimens that enhance antigen-specific T cell reactivity against cancer.

Disclosures

No relevant conflicts of interest to declare.
Thrombomodulin Gene Mutations Associated with Thromboembolic Diseases
Authors: Wang Qingyun , Tang Liang , Hu Bei , Hu Yu , Liu Han
Published: October 2019

Background

Thrombomodulin (TM), encoded by the THBD gene, is an endothelial transmembrane glycoprotein that acts as a cofactor for thrombin in the activation of protein C (PC). It has been clearly demonstrated that the anticoagulant and profibrinolytic functions of the thrombin-TM-PC system play critical roles in the prevention of thromboembolic diseases.

Methods and Results

Using direct sequencing, six novel mutations were identified in THBD gene of eight patients with either arterial or venous thrombosis: c.376G>T (p.Asp126Tyr), c.569C>G (p.Ser190Trp), c.659T>G (p.Leu220X), c.1208G>A (p.Arg403Lys), c.1288G>A (p.Gly430Ser), c.*6_*23del and c.*23_*40del. Of the mutations, c.376G>T and c.*6_*23del were detected in a 10-year old child with venous thrombosis (VTE), who died one year later. The c.376G>T point mutation predicting p.Asp126Tyr in thrombomodulin protein was also detected in a 38-year old VTE patient. The nonsense mutation c.659T>G recurred in a 55-year old patient with coronary artery disease (CAD) and a 46-year old VTE patient, who died from repeated venous thrombosis two years later. The wild type and the mutant thrombomodulin were expressed in HEK-293t cells. Two mutations were found to have reduced TM protein expression compared to wild-type (100%), Ser190Trp (69.1% ±5%, P < .05), p.Leu220X (19.1% ± 7.6%, P < .001). Comparing to wild type, four point mutations diminished the cofactor activity of TM according to the reduced level of activation of PC in cultured cells. As for the two 18-bp deletions, a psiCHECKTM-2 vector containing wild type and mutant 3'UTR of TM was constructed. Compared with the wild-type, the deletions significantly reduced the reporter gene-expression level. The antigen level and activity in cultured cells with c.376G>T have no significant difference with the wild type, indicating that the mutant may cause a thrombophilic state through other unknown pathological mechanisms.

Conclusion

The results suggest that gene mutation of thrombomodulin may be important in the pathogenesis of thrombotic diseases. Further study on genetics of thrombosis should focus on resequencing of THBD gene in different populations.

Disclosures

No relevant conflicts of interest to declare.
Thyroid Hormone Receptor Beta and NCOA4 Regulate Terminal Erythrocyte Differentiation
Authors: Lee Hsiang-Ying , Lodish Harvey F. , Gao Xiaofei , Li Wenbo , Platt Randy , Barrasa Inmaculada , Elmes Russell , Rosenfeld Michael G.
Published: October 2019

An effect of thyroid hormone on erythropoiesis has been known for more than a century but the molecular mechanism(s) by which thyroid hormone affects red cell formation are still elusive. Our study demonstrates an essential role of thyroid hormone during terminal human erythroid cell differentiation; specific depletion of thyroid hormone from the culture medium completely blocked human erythroid differentiation. Genome wide analysis showed that thyroid hormone receptor β (TRβ) occupies many gene loci encoding transcripts abundant during terminal erythropoiesis, including globin genes, and cooperates with GATA-1 and RNA polymerase II (Pol II) to regulate their expression. Treatment with TRβ agonists enhanced erythroblast differentiation in vivo and alleviated anemic symptoms in a chronic anemia mouse model. To identify factors that cooperate with TRβ during human erythroid terminal differentiation, we conducted RNA-Seq in human reticulocytes and identified NCOA4 as a critical regulator of terminal differentiation. Furthermore, Ncoa4-/- mice are anemic in both the embryonic and perinatal periods and fail to respond to thyroid hormone by enhanced erythropoiesis. Genome wide analysis suggests that thyroid hormone promotes NCOA4 recruitment to chromatin regions that are in proximity to Pol II and are highly associated with transcripts abundant during terminal differentiation. Collectively, our results reveal the molecular mechanism of thyroid hormone function on red blood cell formation and are potentially useful to treat certain anemias.

Disclosures

No relevant conflicts of interest to declare.
A Novel Isoflavone, ME-344, Targets the Cytoskeleton in Acute Myeloid Leukemia
Authors: Jeyaraju Danny V , Hurren Rose , Wang Xiaoming , MacLean Neil , Gronda Marcela , Jordan Craig T , Minden Mark D. , Giaever Guri , Schimmer Aaron D.
Published: October 2019

The isoflavone ME-344 is a potent anti-cancer agent with preclinical efficacy in solid tumors in vitro and in vivo. In a recently completed phase I clinical trial in patients with refractory solid tumors, ME-344 was well tolerated and clinical responses in refractory patients were observed. ME-344 has been shown to reduce mitochondrial ATP generation in a tumor selective manner, though other potential activities have not been fully defined. In addition, the preclinical efficacy of ME-344 in leukemia has not been established. Therefore, we investigated the anti-leukemic properties and the mechanism of action of ME-344.

We treated a panel of 7 leukemia cell lines with increasing concentrations of ME-344, and measured cell growth and viability. ME-344 was cytotoxic to the 7 leukemia cell lines with an IC50 in the range of 70-260 nM. In addition, ME-344 induced preferential death in primary AML patient samples over normal hematopoietic cells. In an OCI-AML2 xenograft model, ME-344 reduced tumor growth by up to 95% of control without evidence of toxicity.

Mechanistically, as reported in studies in solid tumors, ME-344 increased mitochondrial ROS generation in leukemic cells. However, antioxidant treatment did not rescue cell death, suggesting that ME-344 has additional targets beyond the mitochondria. To identify additional targets of ME-344, we conducted haplo-insufficiency profiling in S.cerevisiae to identify genes whose heterozygous deletion confers increased sensitivity to ME-344. The top hits from the screen were genes involved in cytoskeletal organization. Therefore, we tested the effects of ME-344 on actin and tubulin polymerization in cell free assays. While ME-344 did not inhibit actin polymerization, it inhibited tubulin polymerization by interacting near the colchicine-binding site and at a site distinct from vinca alkaloids. Furthermore, cells resistant to tubulin inhibitors due to tubulin point mutations were also resistant to ME-344. Finally, we showed that ME-344 synergizes with vinblastine in leukemia cells. Thus, our study demonstrates that ME-344 displays preclinical efficacy in leukemia through a mechanism at least partly related to targeting tubulin polymerization.

Disclosures

No relevant conflicts of interest to declare.
SNPs in IL-10, TNF-Alfa, and NFkB1 Genes and Their Association with Prognostic Parameters in Patients with Hodgkin Lymphoma
Authors: Gaiolla Rafael Dezen , Moraes Marcelo Padovani de Toledo , de Oliveira Deilson Elgui
Published: October 2019

Hodgkin lymphoma (cHL) is a malignant lymphoid neoplasia that shows aberrant expression of cytokines, including IL-10 and TNF-a. Single nucleotide polymorphisms (SNPs) in genes encoding IL-10 and TNF-a or their regulatory proteins, such as NFkB, may impact cHL pathobiology. In this study, SNPs in the promoter regions of genes encoding for IL-10 (SNP/pIL-10 -592, rs1800872; and SNP/pIL-10 -1082, rs1800896), and TNF-a (SNP/pTNF-a -238, rs361525; and SNP/pTNF-a -862, rs1800630), as well as in the intronic region of the NFkB1 gene (SNP/iNFkB1, rs1585215) were genotyped in 73 patients with cHL and evaluated against clinical and laboratory prognostic parameters for the disease. SNPs/pIL-10 AA were significantly associated with higher leukocyte and lower lymphocyte counts at diagnosis. In case of TNF-a, SNP/pTNF-a -238 AG was associated with EBV infection while SNP/pTNF-a -862 CC presented more frequently with leukocytosis. SNP/iNFkB1 AA generally had stage IV and extranodal disease at diagnosis. Nonetheless, none of the studied SNPs had effect on treatment outcome. This study shows that some SNPs genotypes for IL-10 and TNF-a genes are associated with prognostic parameters in cHL; furthermore, this is the first time that the SNP/iNFkB1 (rs1585215) was implicated in clinical features of the disease.

Disclosures

No relevant conflicts of interest to declare.
ID3 Loss In Vivo Synergizes with MYC-Driven Oncogenesis in Burkitt Lymphoma
Authors: Love Cassandra , Remple Rachel , Li Guojie , Dave Sandeep
Published: October 2019

Burkitt lymphoma (BL) is an aggressive cancer, characterized by the deregulation of the c-myc (MYC) gene. BL is a major model disease for molecular studies in many other malignancies. Treatment of BL requires long courses of intensive chemotherapy which extracts a disproportionate toll on patients.

Previous studies using next generation sequencing have identified ID3 as a tumor suppressor gene, with silencing mutations in about 40% of BLs. However, relatively little is known regarding the biological role of ID3 in BLs.

In this study, we sought to elucidate the molecular mechanisms that are affected by ID3 loss in BL. We utilized two complementary strategies. First, we developed an in vivo approach through conditional knockout of Id3 in mice. Second, we utilized a CRISPR-based approach to genetically knockout ID3, to study the effects of ID3 loss in BL cell lines.

We have successfully knocked out exon 1, including the functional helix-loop-helix (HLH) domain of ID3 in 4 BL cell lines (Namalwa, Ramos, BJAB, and Jijoye) that do not harbor mutations in ID3. Deletion of exon 1 causes complete deletion at the protein level. Transcriptome sequencing from ID3 mutated BL cases identified increase in the expression of MYC target genes, as well as proliferation-related genes. Cell cycle analyses revealed faster cell cycle progression in the ID3 knockdown cells (P=0.02).

To study ID3 loss in vivo, we crossed Em-Myc+ mice with a standard Id3 knockout strain mice. We found that E m -Myc+;Id3-/+ mice demonstrated reduced latency to tumor development compared to Eµ-Myc+;Id3+/+ littermates (P<0.01). This decreased tumor latency noted in the E m -Myc+;Id3-/+ mice suggests a strong in vivo synergy between the oncogenic effects of MYC overexpression and ID3 loss. Many of these tumors displayed the classic "starry sky" pattern that is classic for human Burkitt lymphomas. We are continuing to use the AID -Cre positive strains to generate combined Id3 deletion and Myc over-expression specifically in germinal center B cell context. Additional characterization of the mouse tumors using mouse whole exome sequencing and transcriptome sequencing is in progress and will be presented in the meeting.

Our data reveal that ID3 greatly potentiates the oncogenic role of the MYC gene and understanding the role of ID3 enables new insights into the pathogenesis of BL, and potentially other MYC-driven malignancies.

Disclosures

No relevant conflicts of interest to declare.
Distrust and Conflict in Sickle Cell Disease: Intersecting Narratives of Patients and Physicians
Authors: Tsyvkin Elina , Riessman Catherine , Mathew Paul
Published: October 2019

A gap in trust between patients with sickle cell disease (SCD) and medical providers is well recognized, largely originating from repetitive requests for opioid analgesics. Although expert clinical care guidelines in sickle cell disease are available, they rarely address measures by which this endemic gap in trust may be narrowed to fulfill the goals of medicine. We hypothesized that an increased familiarity with the patient as an individual through exposure of physicians to first-person narratives of the life-world of SCD may allow reformulation of perceptions and narrow the gap between physicians and patients. In a pilot study, extended first-person narratives of the illness experience elicited from patients with SCD with a history of recurrent hospitalizations (n=10) point to the individualized impact of pain, illness and stigmatizing disruptions in life-building efforts imposed by SCD together with the recurrent conflicts with medical caregivers, particularly after transition from pediatric to adult medicine. Patient-elicited narrative fragments such as "You are constantly fighting with people who are supposed to be making us feel better", "You don't know me, I am a church boy!" and "Put down your microscopes and talk to us" reveal the yearning by patients for individual integrity to be acknowledged, absorbed and interpreted by physicians. Additional narrative fragments such as "I hate myself. I hate my life", "Why am I broken?", "you don't want to be the girl with jaundice", "I had to leave work for 2 months because I was hospitalized, it killed me, it killed me", "you never know when it's going to be the last day, the last moment", exhibit the woundedness and fragility of existence experienced by patients. Narratives elicited separately from physicians caring for SCD (n=5) reveal anxiety, fear and distrust of the reported pain experience and opioid requests, and point toward deeper schisms that disparities may define: "I was very scared when I took care of my first sickle cell patient", "I came in with a bias already", "a similar frequent flier who had the same kind of behavior", "you have to be careful with empathy, people can take advantage of it", "I am not an enabler", "My grandfather told me about these people". Yet when the same physicians were invited to read and reflect on transcripts of the life-world stories of patients, reshaped perceptions of the stigmatized patient are revealed; "without knowing the story, you can't put it all together", "this would change the way I view treating them", "I will have more patience now" as examples. However, "Why can't they listen to our story? It could help them to be better patients," suggests that additional studies of the mutual intersections of patient and physician narratives are warranted and that these can offer insights and pathways toward mitigating distrust and conflict between medical care providers and patients with SCD.

Disclosures

No relevant conflicts of interest to declare.