bluuebunny/crossref_metadata_2025 huggingface dataset:
Higher MMP9 and FGF2 Protein Levels and Elevated MMP9/TIMP Ratios Were Associated with JAK2V617F Mutation Regardless of Allele Burden in Myelofibrosis
Authors: Lima Luciene Terezina , Nunes Daniela Prudente Teixeira , Gomes Guilherme Wataru , Chauffaille Maria de Lourdes L. F. , Silva Maria Regina Regis , Ponce Cesar Cilento , Castro Fabiola Attié , Santos Marcos Tadeu , Guerra-Shinohara Elvira M
Published: October 2019

Background: Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are involved in tumor neoangiogenesis which plays an important role in hematological neoplasias progression and prognosis. Myelofibrosis (MF) and essential thrombocythemia (ET) are myeloproliferative neoplasms (MPNs) resulting from acquired hematopoietic stem cell mutations. Most patients share the JAK2V617F mutation and may exhibit substantial overlap. The molecular mechanisms involved in MPNs pathogenesis are not completely elucidated. In this context, evaluation of genes and proteins related with angiogenesis could be useful to better understand MPN pathobiology.

Aims: To investigate the mRNA and protein expressions of MMPs and TIMPs and their relationship with plasma markers of angiogenesis (VEGFA and FGF2) and JAK2V617F mutation status in MF and ET patients.

Methods: Forty-three MPNs patients (MF=29 and ET=14) and 36 control subjects were studied. The controls were matched with patients according to age and gender. MPN diagnosis was defined according to the 2008 WHO criteria. The MMP2, MMP9, TIMP1 and TIMP2 mRNA expression from peripheral leukocytes and serum proteins levels were measured by real time PCR and Luminex technology. VEGFA and FGF2 concentration were measured in plasma from MPN patients and controls. JAK2V617F mutation status and allele burden and MPL W515K/L analysis were performed in total leukocytes DNA by real time PCR, using Taqman MGB probes.

Results: Higher MMP2, MMP9 and TIMP1 mRNA expression were observed in MF patients compared to controls (P<0.05). The frequencies of JAK2V617F mutated patients were 62.1% for MF and 35.7% for TE. The allele burden was similar in MF and ET groups. MF patients positive for JAK2V617F presented higher protein levels of MMP9 (N=18, P=0.008) and FGF2 (N=18, P=0.009) when compared with non-carriers of mutation (N=11). In contrast, no correlations were found between allele burden and MMP9 (r=0.153, P=0.558, N= 18) or allele burden and FGF2 (r=0.069, P=0.794, N=18). Carriers of JAK2V617F in MF group (N=17) showed elevated MMP-9/TIMP1 and MMP9/TIMP2 ratios of protein levels than those MF JAK2V617F negative (N=11, P<0.025). ET patients JAK2V617F positive presented higher MMP9 mRNA level (N=5) compared with negative ones (N=9, P=0.009), but no difference was observed in protein levels. None of patients presented MPL W515K/L mutation.

Conclusions:Our findings suggest that MF patients with JAK2V617F mutation exhibit higher MMP9 and FGF2 protein levels and elevated MMP9/TIMP ratios, which are not associated with JAK2V617F allele burden.

Financing: FAPESP 2012/12957-5

Disclosures

No relevant conflicts of interest to declare.
Effect of Hes1 on AML Cells: A Potential Therapeutic Target
Authors: Tian Chen , Jia Yongsheng , Hu Dongzhi , Zhang Yizhuo
Published: October 2019

Notch signal pathway is an important mediator of growth and survival in several cancer types, while Notch pathway genes function as oncogenes or tumor suppressors in different cancers. Although aberrant Notch activation contributes to leukemogenesis in T cells, the role of Notch pathway in acute myeloid leukemia (AML) remains unclear. To address this problem, we investigated the expression levels of its downstream effector Hes1 and p21 in primary AML cells and cell lines by realtime PCR and western, and found that both of them was weak in these cells. Induced activation of Hes1 consisently led to AML growth arrest and apoptosis, which was associated with enhanced p21 expression. Besides, overexpression of Hes1 inhibited growth of AML cells in vivo. In conclusion, we reported that Hes1 mediated growth arrest and apoptosis of human AML cells, and demonstrated a novel tumor suppressor role for Hes1 in AML.

Disclosures

No relevant conflicts of interest to declare.
The Experience of the International Registry for Chronic Myeloid Leukemia (CML) in Children and Adolescents (I-CML-Ped Study): Pronostic Consideration
Authors: Millot Frédéric , Guilhot Joelle , Suttorp Meinolf , Sedlacek Petr , de Bont Eveline S.J.M. , Li Chi Kong , Kalwak Krzysztof , Lausen Birgitte , Srdjana Culic , Dresse Marie-Francoise , Biondi Andrea , Baruchel Andre
Published: October 2019

Aims: An international registry (I-CML-Ped Study)was established to assess epidemiology, management and outcome of CML in the pediatric population.

Methods: All national pediatric study groups were invited to include newly diagnosed children and adolescents less than 18 years with CML diagnosed later than January 2000.

Results: Since January 2011, 351 patients from 12 countries have been registered. Clinical and biological data at initial diagnosis are available in 278 patients.There was a male preponderance (57%). The median age at diagnosis was 12.4 years (range, 9 months -17.5 years); 6% of the patients were younger than 4 years. At time of diagnosis 92% of the children were in chronic phase, 8% in accelerated phase and 1% in blastic phase according to the European Leukemia Net criteria. The Sokal risk group distribution was: 18% low, 31% intermediate and 51% high risk. The majority of the patients showed a Lansky score of 100 (59%) or 90 (21%). Splenomegaly was present in 76% of patients. The median of the spleen size below the costal margin was 11 cm (range: 1 to 25 cm). The median of the leukocyte count was 235x109/L (range: 5 to 1038). Additional chromosomal abnormalities in Ph-positive cells were observed in 6 % of the patients. The BCR-ABL transcript type was available in 227 patients: b3a2 54%, b2a2 38%, b3a2-b2a2 6% and b2a3 2%. The median follow-up time is 39 months (range, 0.5-161). Eight deaths were recorded. The estimated overall survival rate at 60 months is 95% (95%CI 89-97). Irrespective of treatment and follow-up, 124 (73%) of 169 assessable patients for cytogenetic response achieved complete cytogenetic response (CCyR). Exploratory analyses were performed in newly diagnosed patients regarding clinical and biological factors influencing the achievement of CCyR 12 months after starting imatinib. In univariate analyses, the Eutos score, the spleen size, the hematocrit level, the lymphocyte count and immature cells in peripheral blood, the percentage of granulocytes and monocytes in the marrow were identified as potential prognostic factors. However, only the percentage of the granulocytes in the marrow was identified as independent factor of achievement of CCyR at 12 months in multivariate analysis. Data collection and quality control regarding molecular assessment are ongoing.

Conclusion: The data indicates that children and adolescents with CML presented with clinical and biological differences compared to adult patients with CML. Identification of prognostic factors is needed to optimize the strategy in the pediatric population.

Acknowledgment: The I-CML Ped study is supported by an unrestricted grant from Novartis Pharmaceutical Company

Disclosures

No relevant conflicts of interest to declare.
Artesunate Enhanced Apoptosis of Human High Risk MDS Cells Induced By the DNMT Inhibitor Decitabine
Authors: Ying Wang , Zhiyun Niu , Shupeng Wen , Fuxu Wang , Xuejun Zhang
Published: October 2019

Abstract: The present study evaluated whether the artesunate (ART) could enhance the apoptosis induced by decitabine (DAC) in SKM-1 cells, and to explore its potential mechanisms. Cytotoxicity of the combination of ART and DAC on SKM-1 cells was detected by CCK-8 assay and the apoptosis rate of high-risk myelodysplastic syndrome (MDS) cell line, SKM-1 cells was measured by flow cytometry. Protein expression levels of activated caspase-3, caspase-9, caspase-8, cleaved PARP and AIF in SKM-1 cells were measured by Western blot. Laser confocal microscope analysis showed AIF transfer to the nucleus. The growth inhibition rate and the apoptosis rate of SKM-1cells treated with the combination of ART and DAC was significantly increased compared with that of SKM-1 cells treated with the single agent (P<0.05). Both ART and DAC could induce caspase-dependent apoptosis, while ART, but not DAC, also induced caspase-independent apoptosis via AIF transfer from mitochondria to the nucleus. Additionally, the combination of the two agents induced cell death was not attenuated by caspase3,7 inhibitor Ac-DEVD-CHO. Our result suggested that the ART–DAC combination was more effective than single-agent therapy in vitro. Both compounds not only activated caspase-dependent pathway but also activated a caspase-independent mitochondrial pathway.

Disclosures

No relevant conflicts of interest to declare.
Establishment of TRAIL-Resistance Kasumi-1 Cell Line and the Analysis of It different mRNA Expression Profile with the Original Kasumi-1 Cell Line
Authors: Jiang Xuewei , Zengkai Pan , Jin Chen , Pengfei Yu , Liu Li-Gen
Published: October 2019

Introduction

Tumor necrosis factor related apoptosis inducing ligand (TRAIL) can induce the apoptosis of many human leukemia cells while sparing of normal cells, but its resistance is also universal. Our previous study on apoptosis of t(8;21) positive acute myeloid leukemia cell line Kasumi-1 induced by rhTRAIL showed that the survival rate no longer decreased significantly when rsTRAIL reached a certain concentration which implied Kasumi-1 cells might have a resistant tendency to TRAIL. Then, we established a TRAIL-resistant Kasumi-1 cell line (Kasumi-1 TR) by intermittently escalating rsTRAIL concentration in culture media, and compared the mRNA expression profile with the original Kasumi-1 cell line by using Affymetrix Human Genome U133 Plus 2.0 Array.

Methods

Kasumi-1 TR cell line was established by intermittently treated Kasumi-1 cells with progressively escalating rsTRAIL concentration. Proliferation of leukemia cells were measured by CCK-8 assay, and rsTRAIL IC50 of cells and resistance index were calculated according to proliferation of cells treated with rsTRAIL at different concentrations. TRAIL and TRAIL receptors 1-4 on cells surface were detected by flow cytometry. Expression profiles of Kasumi-1 cells and Kasumi-1 TR cells were analyzed by Affymetrix Human Genome U133 Plus 2.0 Array to identify differentially expressed genes, and the search of genes possibly related with TRAIL-resistance were using by GO functional analysis and pathway enrichment analysis.

Results

1) Kasumi-1 TR cells proliferation was faster than that of Kasumi-1 cells(Fig 1A); 2) IC50 of 24 hours for Kasumi-1 cells was 756.833ng/ml (logIC50 2.879 ± 0.148), IC50 of 24 hours for Kasumi-1 TR cells was 1634646.005ng/ ml (logIC50 6.213 ± 0.637), the RI of 24h was 2159 (Fig 1B); IC50 of 48 hours for Kasumi-1 cells was 345.390ng/ml (logIC50 2.538 ± 0.153), IC50 of 48 hours for Kasumi-1 TR cells was 33642.641ng/ml (logIC50 is 4.257 ± 0.317), the RI for 48h was 97 (Fig 1C); 3) Cell surface expression of TRAIL and its receptors 1-4 had no difference between two cell lines(Fig 1D). 4) There were 1537 genes up regulated by more than 2 times while 487 genes down regulated by more than 2 times in Kasumi-1 TR cells compared with the original Kasumi-1 cells (Fig 1E). Of which BCL-2 family antiapoptotic gene BCL2 is increased by 3.153 times and BCL2A1 increased by 18.23 times, IFNAR1 involved in JAK/STAT pathway increased by 12.841 times and TRAIL death receptor TNFRSF10A down regulated by 3.256 times(Fig 1F).

Conclusions: The Kasumi-1 cell line with rsTRAIL resistance (Kasumi-1 TR) is established, and its resistance may be associated with the up expression of BCL2, BCL2A1, IFNAR1 and down regulated expression of DR4.

Acknowledgment

This work was supported by grants from NSFC (30672415) and STCSM (054119528).

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
The Outcome of Childhood High-Risk Acute Lymphoblastic Leukemia: 10 Years of Experience with the ALL-BFM 95 Protocol in Chinese Single Centre
Authors: Ruan Yongsheng , Wu Xuedong , Li Chunfu , Feng Xiaoqin
Published: October 2019

Objectives: To evaluate the effectiveness and the practicability of Acute Lymphoblastic Leukemia Berlin-Frankfurt-Münster 95 (ALL-BFM 95) protocol in treating childhood high-risk (HR) acute lymphoblastic leukemia (ALL) in Chinese single centre.

Methods: A retrospective analysis of 47 children with novo high risk ALL from July 2003 and August 2013 was performed. These children treated by the ALL-BFM 95 protocol. Median follow-up time was 39 months (16~127months). Survival was evaluated by Kaplan Meier analysis and Log-Rank test.

Results:Relapse-related death occurred in 12 of 47 patients (25.5%), and 5 of 47 patients (10.6%) were treatment-related mortality. Five-year probability of event-free-survival (pEFS) were found to be 62.4%. High risk ALL children only with prednisone poor response(PPR) has good outcome who’s Five-year probability of event-free-survival (pEFS) could reach to 80%. Children with hematopoietic stem cell transplantation (HSCT) after chemotherapy achieved significantly better pEFS than only chemotherapy (77.3% vs 51.6%，p=0.035). pEFS for B-cell precursor and T-cell ALL was 64.5% and 57.1%, respectively（P=0.633）.

Conclusions: ALL-BFM 95 protocol can also improve the outcome of childhood high risk ALL in China. The leading causes of death were relapse. HSCT treatment had better outcome than only chemotherapy in high risk ALL children in CR1 phase.

Disclosures

No relevant conflicts of interest to declare.
The Prevalence of Congenital Trombophilia in Children Treated for Acute Lymphoblastic Leukemia with Thrombosis As a Complication
Authors: Smalisz-Skrzypczyk Katarzyna , Klukowska Anna , Pawelec Katarzyna , Matysiak Michal
Published: October 2019

The aim of the study was to evaluate factors predisposing to thrombosis, especially congenital trombophilia, in children treated for acute lymphoblastic leukemia according to ALL IC BFM 2002 and 2009.

An analysis of 36 cases of thrombosis in 242 children (15%) treated due to ALL in the Department in years 2007-2013 was carried out. Age at onset, location, cause and treatment methods were taken into consideration. All analyzed patients underwent screening for congenital thrombophilia.

The age of patients ranged from 1.5 to 17 years with the median of 8.5 years. Arterial thrombosis occurred in 2 persons (1%), venous thrombosis in 34 (99%) patients. Thromboembolism was usually related to the central venous catheter (n = 31, 91%). Massive thrombosis occurred in 7 patients (20%). Relapses occurred in 3 children.

The most common causes of thrombosis were: the presence of the central catheter (n = 31, 91%) and L-asparaginase treatment (n = 19, 55%). The factor V Leiden mutation was diagnosed in 1 patient, protein C deficiency in 1 patient as well, and elevated levels of factor VIII were detected in 4 cases.

The mean concentrations of factor VIII, von Willebrand factor and antithrombin were not different in the groups with and without thrombosis (table 1 ). The concentration of fibrinogen measured after finishing the chemotherapy was similar in both groups.

Conclusions 1. A risk factor for thrombosis in children with ALL is the treatment of L-asparaginase and the presence of central venous catheter.

2. Screening for congenital thrombophilia in children treated for ALL should not be done routinely because the coexistence of this two conditions is rare. Table 1FVIII (%)Fibrinogen ( g/l)AT (%)Patients with thrombosis1012,776Patients without thrombosis892,4382p0,45 ( ns)0,2 (ns)0,34 (ns)

Disclosures

No relevant conflicts of interest to declare.
Low Efficacy and High Mortality Associated with Clofarabine Treatment of Patients with Acute Myeloid Leukemia and Higher-Risk Myelodysplastic Syndromes
Authors: Roberts Daniel A , Wadleigh Martha , McDonnell Anne , DeAngelo Daniel J , Stone Richard M. , Steensma David P.
Published: October 2019

Background: Clofarabine, a second-generation nucleoside analogue, has been shown in prospective studies to have activity in relapsed or refractory acute myelogenous leukemia (AML) and higher-risk myelodysplastic syndromes (MDS). However, there are few data evaluating performance of the drug in populations of patients not enrolled in clinical trials.

Methods: We reviewed clinical outcomes for 84 patients treated at our center off-study for relapsed or refractory AML (n=81) or MDS (n=3) with clofarabine as monotherapy (n=19) or in combination with cytarabine (n=65). Almost all patients (n=62) receiving combination therapy were administered cytarabine 1 g/m2/day on days 1-5 and clofarabine 40 mg/m2/day on days 2-6 (Faderl et al Blood 2005; 105:940), while monotherapy dosing ranged from 15-30 mg/m2/day x 5 days.

Results: Median age of patients was 51 years; 54% were male. The median prior salvage regimens received was 1 (range, 0-5). Using International Working Group (IWG) response criteria, the overall response rate (ORR) of all treated patients was 21%, with a complete response rate with either complete or incomplete hematopoietic recovery (CRR = CR + CRi) of 14%. For combination therapy, ORR was 22% with CRR of 18%, and monotherapy patients had an ORR of 21% with CRR of 11%. Although limited by small numbers, subgroup analysis did not reveal significant variation in response rates when comparing age, performance status, or prior number of salvage regimens. 30-day mortality or induction death for all patients, combined therapy, and monotherapy was 21%, 23%, and 16% respectively. Overall median survival for all patients was 3 months; a subset of 12 patients who were able to go to allogeneic stem cell transplant had an 18-month median survival.

Conclusion: Clofarabine’s efficacy in a "real-world" setting appears to be less than has been reported in clinical trials, and treatment is associated with a high early mortality rate. Clofarabine's utility in relapsed/refractory AML/MDS may be primarily for cytoreduction in patients who have an allogeneic transplant option.

Disclosures

Off Label Use: Clofarabine- a second-generation nucleoside analogue approved for pediatric acute lymphoblastic leukemia..
Gene Expression Profiling in the Classification of Acute Leukemia Brazilian Patients
Authors: Melo Carolina Pereira de Souza , Campos Catharina Brant , Dutra Alvaro Pimenta , Atalla Angelo , Dudeque Pianovski Mara Albonei , Carbone Edna Kakitani , Quintão Lares Luciana , Souza Hélio Moraes , Silva Shirlei Octacilio , Ferreira Alessandro Clayton de Souza , Assumpcao Juliana Godoy
Published: October 2019

In the past few decades, genetic data has become increasingly important for acute leukemia diagnosis and patients stratification. Indeed, the present World Health Organization (WHO) leukemia classification system is largely based upon genetically defined subgroups. Gene expression profile (GEP) may correctly predict most genetic leukemia subtypes, but so far no GEP report has evaluate patients from Latin America. In the present study, we used gene expression microarray data to build an acute leukemia classifier. Bone marrow samples were collected from 231 individuals at diagnosis, 110 presented de novo acute myeloid leukemia (AML), 97 had de novo acute lymphoid leukemia (ALL) and the remaining 24 were controls who had other conditions including chronic leukemias or non-hematological diseases. GEP was evaluated based on mRNA expression signatures obtained with the Sure Print G3 Human GE (60k) system (Agilent Technologies). k-nearest neighbors prediction algorithm was applied and the top 60 informative genes were selected for each of the most prevalent genetic subtypes (T-ALL, B-ALL BCR-ABL, B-ALL ETV6-RUNX1, B-ALL TCF3-PBX1, AML PML-RARa, AML RUNX1-RUNX1T1, AML FLT3-ITD, AML NPM1mut). The less prevalent groups such as MLL rearranged and CBFB-MYH11 were not included in the classifier because of the low number of patients carrying these aberrations in our cohort. Performance of each prediction model was assessed by leave-one-out crossvalidation through the GenePattern platform (Broad Institute). The average classifier accuracy was 94.75%. Higher accuracy and precision were achieved for T-ALL (99%/96%) and AML PML-RARa (97%/97%). However, for ALL BCR-ABL, AML FLT3-DIT and AML NPM1mut the gene signature had low precision rates (74%, 66% and 80%, respectively). The data presented here confirm that a single platform of gene expression followed by bioinformatic analysis can correctly classify genetic subgroups, but a refinement of the classifier developed is needed in order to improve the detection of heterogeneous entities such as BCR-ABL or FLT3-ITD carriers.

Disclosures

No relevant conflicts of interest to declare.
Predictive value of therapeutic monitoring with plasma Epstein-Barr virus DNA in Extranodal NK/T cell lymphoma, nasal type
Authors: Guo Chengcheng , Huang He , Tian Ying , Li Xueying , Zhang Mengping , Hong Huangming , Fu Xiaohong , Lin Xiaoting , Li Shanshan , Prem Raj Shrestha , Xia Zhongjun , Lin Tongyu
Published: October 2019

Predictive value of therapeutic monitoring with plasma Epstein-Barr virus DNA in Extranodal NK/T cell lymphoma, nasal type

Background: Predictive tumor markers are essential for extranodal NK/T cell lymphoma, nasal type (ENKTL), which pursues an aggressive clinical course with poorer prognosis. This prospective study was conducted to evaluate the dynamic monitoring value of circulating EBV DNA concentration for the prediction of ENKL during treatment.

Method: Patients with untreated, centrally reviewed diagnosis of ENKTL were included in our study. Plasma Epstein-Barr virus (EBV) DNA levels were collected before and/or every 2 cycles of chemotherapies for circulating EBV DNA measurement by a real-time PCR assay.

Result: From Jan 2002 to Aug 2012, 113 newly diagnostic ENKTL patients enrolled. The positive rate of circulating EBV DNA is 61.9% (70/113) with a median concentration of 1.21×103copies/ml. Patients who had initial positive circulating EBV-DNA had more lymph nodes invasion, higher IPI and KPI score. The more advance of the stage, the higher rate of the circulating EBV-DNA positive. When divided the positive group as low and high-dose ones by the cut-off value as 2×104copies/ml, the CR rate of the high-dose group is much lower and the 5-year OS is significantly better than the low-dose group and the negative group. After two cycles of chemotherapy, 45 patients were tested circulating EBV-DNA and 53.33% (24/45) patients became negative EBV-DNA candidates. There were 87.5% (21/24) patients obtained complete remission at the end of the treatment, comparing as 42.86% (9/21) in the still positive EBV-DNA groups(P= 0.002). There is a tendency that the patients whose circulating EBV-DNA become negative after two cycles will have better prognosis (5-year OS: 75% vs 46%, P=0.074). The similar situation of CR rate and OS happened in the EBV-DNA detection of completion of total chemotherapy. 7 of 13 relapsed pts of ＞1×103copies/ml EBV-DNA at the time of recurrent, and the survival outcome was dismal for them compared to the other 6 pts of ≤1×103copies/ml（5- year OS: 0% vs 80%, P=0.001）

Conclusion: Circulating EBV-DNA level can predict the efficacy of treatment as a dynamic marker of ENKL. Patients with early positive detection of EBV-DNA after 2 cycles of chemotherapy maybe received more aggressive treatments.

Disclosures

No relevant conflicts of interest to declare.
Effect of High Body Mass Index on the Prognosis of Diffuse Large B-Cell Lymphoma
Authors: Hattori Yukako , Tomita Naoto , Suzuki Taisei , Ishiyama Yasufumi , Kishimoto Kumiko , Koyama Satoshi , Takahashi Hiroyuki , Miyashita Kazuho , Nakajima Yuki , Motohashi Kenji , Takasaki Hirotaka , Kawasaki Rika , Hashimoto Chizuko , Fujimaki Katsumichi , Sakai Rika , Fujisawa Shin , Motomura Shigeki , Ishigatsubo Yoshiaki
Published: October 2019

Background

Obesity has been pointed out as one of the risk factors for the development in several neoplastic diseases including malignant lymphoma. However the impact of obesity on the outcome of malignant diseases is unclear. L.Weiss et al have shown that high body mass index (BMI) is a significantly better prognostic factor in diffuse large B-cell lymphoma (DLBCL) (Increased body mass index is associated with improved overall survival in diffuse large B-cell lymphoma. Annals of Oncology 2014; 25: 171-176.). We evaluated this hypothesis in Japanese patients with DLBCL.

Patients and Methods

We analyzed 338 patients with newly diagnosed DLBCL who received full-dose (80% or more of the prescribed dose) R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) between April 2003 and December 2009 across 7 institutes. Patients of all stages were treated 6 or 8 cycles of full-dose R-CHOP therapy. All the patients were classified into 2 groups: high BMI (≥25 kg/m2 ) or low BMI (<25 kg/m2).

Results

The median patient age was 65 years (19–80 years); 192 were men and 146 were women. The median BMI was 22.9 kg/m2 (14.1–40.4 kg/m2); 66 patients (19.5%) were in the high BMI group (median, 27.4 kg/m2) and 272 (80.5%) were in the low BMI group (median, 21.6 kg/m2). The median follow-up of patients who were alive (n = 271) was 67.5 months (12.6–121.8 months). The 5-year progression-free survival (PFS) and overall survival (OS) of the high versus low BMI groups were 63.9% versus 74.0% (P = 0.196) and 77.2% versus 84.2% (P = 0.230), respectively (Figure1,2). Multivariate analysis considering the sex, International Prognostic Index, B symptoms, bulky mass, and BMI showed that high BMI (≥25 kg/m2) was a significant adverse prognostic factor for both OS (hazard ratio [HR], 1.906; 95% confidence interval [CI], 1.08-3.376; P = 0.027) and PFS (HR, 1.710; 95% CI, 1.054-2.774; P = 0.030).

Conclusion

The results of our study indicated that high BMI was an adverse prognostic factor of DLBCL. It is necessary to investigate BMI and its influence on patient background or treatment in much more cases.

Figure 1 Figure 1.

Figure 2 Figure 2.

Disclosures

No relevant conflicts of interest to declare.
Plasma DNA Methylation of p16 and shp1 in Patients with B-Cell Non-Hodgkin Lymphoma
Authors: Fu Rong , Chen Xiaoshuang , Shao Zonghong
Published: October 2019

To discuss the clinical implications of cell free DNA, methylation status of the p16 and shp1 genes in plasma DNA from patients with B-NHL were analyzed. Methylation specific polymerase chain reaction (MSP) was used to detect the methylation status of these genes in plasma, peripheral blood leukocytes (PBLs), and formaldehyde-fixed, paraffin-embedded (FFPE) tumor tissues of 73 B-NHL patients. Results showed methylation frequencies of p16 in plasma, PBLs, and FFPE tumor tissues of B-NHL patients were 37%(27/73), 16% (12/73), 39% (16/41); Whereas those of shp1 were 47% (34/73), 25% (18/73), 63% (26/41). High methylation consistencies of p16/shp1 were observed between plasma and FFPE tumor tissues. In plasma and FFPE tumor tissues, there was a higher frequency of methylated p16 in B-NHL patients who had later disease stage, while no similar association was observed in PBLs samples; Higher frequency of methylated p16 was observed in B-NHL patients who had B symptoms and lower platelet count (<100×109/L) in all three samples. Methylated shp1 was frequently detected in patients with high serum LDH level. In summary, DNA methylation in plasma may be promising biomarkers for B-NHL diagnosis, prognosis evaluation, and targeted therapy.

Disclosures

No relevant conflicts of interest to declare.
The Analysis of Prognostic Significance of Absolute Lymphocyte Count and Lymphocyte-to-Monocyte Ratio in Diffuse Large B-Cell Lymphoma
Authors: Huang Haiwen , Liu Ying , Jin Zhengming , Depei Wu
Published: October 2019

【Objective】

To evaluate the prognostic value of absolute lymphocyte count (ALC)and lymphocyte-to-monocyte ratio (LMR) in Diffuse large B-cell lymphoma(DLBCL) when they are diagnosised.

【Methods】

Sixty-nine cases were analyzed retrospectively, including sex, age, stage, B symptoms, absolute lymphocyte count, absolute monocyte count, lymphocyte-to-monocyte ratio, lactate dehydrogenase, IPI score, bulky disease, ECOG performance status, number of extranodal involvement. Thirty-four received CHOP treatment (cyclophosphamide, vincristine, adriamycin, dexamethasone) , thirty-five received R-CHOP (rituximab plus CHOP). The ALC, AMC, LMR cutoff value for survival analysis were 1.0×109/L, 0.3×109/L, 2.6 respectively. All data were analyzed by statistical package for the social sciences (SPSS) software ‘‘version 18.’’ Chi-square test wasused to compare the difference between two groups of means. The Kaplan–Meier method was used to summarize OS and PFS and the logrank test was used for univariable analysis. The Cox proportional hazards model was used for multivariable analyses of measured factors.

【Results】

1. Patients with ALC<1.0×109/L had a high incidence of advanced Ann Arbor stage (P=0.003) , B symptoms (P=0.001) , elevated LDH level (P<0.001), high IPI score (P<0.001) and high ECOG score (P=0.001). Similar results were observed in patients with LMR<2.6. Patients with LMR<2.6 had a high incidence of advanced Ann Arbor stage (P<0.001) , B symptoms (P=0.005) , elevated LDH level (P<0.001), high IPI score (P<0.001) and high ECOG score (P=0.006).

2. In comparison with CHOP and RCHOP treatment, OS was significant longer with rituximab, as well as PFS(P＜0.05).

3. After a median 41.5 months follow-up, K-M analysis showed that lower ALC and LMR associated with inferior overall survival (OS) and progression free survival (PFS) (P＜0.001). In CHOP treatment group, patients with lower ALC and LMR seemed to have worse OS and PFS ( ALC: P=0.002 in OS,and P=0.005 in PFS; LMR: P=0.011 in OS, and P=0.027 in PFS). We got the same results in RCHOP set (ALC: P=0.001 in OS ; P=0.001 in PFS / LMR: P=0.001 in OS ; P=0.002 in PFS).

4. In multivariate analysis, only ALC were proven as an independent prognosis factor of survival(P<0.05).

【Conclusions】

the ALC and LMR at diagnosis were independent prognostic factors of both OS and PFS for patients with DLBCL. These data suggest that ALC can be used in combination with other prognostic features to better predict the outcome of DLBCL.

Disclosures

No relevant conflicts of interest to declare.
Hepatitis C Infection Is Associated with Hepatic Toxicity but Does Not Compromise the Survival of Patients with Diffuse Large B Cell Lymphoma Treated with Rituximab-Based Chemotherapy
Authors: Bai Li-Yuan , Chen Tzu-Ting , Chiu Chang-Fang , Yang Tse-Yen , Yeh Su-Peng , Liao Yu-Min
Published: October 2019

Introduction: The influence of hepatitis C virus (HCV) infection on outcome of patients with diffuse large B cell lymphoma (DLBCL) treated with rituximab-based chemotherapy is still controversial.

Materials and Methods: We retrospectively analyzed the characteristic and clinical outcomes of patients with DLBCL newly diagnosed between Jan 2005 and Dec 2011. All patients were treated with at least one course of rituximab-containing chemotherapy. The clinical characteristics between groups (29 HCV-positive or 139 HCV-negative) were compared using a Chi-square method for categorical variables, a Student’s t-test for continuous variables, and a non-parametric method (Wilcoxon Sum-Rank test) for AST, AST and total bilirubin. Univariate analysis followed by multivariate analysis was used to identify the variables associated with hepatotoxicity.

Results: The medium follow-up duration was 3.0 (0.07-8.02) years. HCV infection resulted in more hepatic toxicity in both univariate (55.2% vs. 23.0%, P = 0.001) and multivariate (P = 0.003) analyses. In addition, HCV-positive DLBCL patients were more likely to have treatment delay (20.1% vs. 0.7%, P = 0.004). For patients who developed hepatic toxicity during immunochemotherapy, HCV-positive patients had significantly higher folds of aspartate aminotransferase elevation (3.39 folds vs. 2.06 folds, P = 0.042) and total bilirubin elevation (1.60 folds vs. 0.96 folds, P = 0.012) compared with those who were HCV negative. However, HCV did not influence the 5-year progression-free survival rate (P = 0.412) or 5-year overall survival rate (P = 0.410).

Conclusions: HCV infection is associated with increased incidence and severity of hepatic toxicity and delayed chemotherapy without compromised survival in DLBCL patients treated with rituximab-based chemotherapy.

Disclosures

No relevant conflicts of interest to declare.
The Prognostic Significance of Tumor-Associated Macrophages and NK Cells in Patients with Diffuse Large B-Cell Lymphoma Treated with RCHOP
Authors: Xie Muchen , Wei Yongqiang , Yang Lei , Huang Fen , Wei Xiaolei , Jing Hui , Hao Xiaoxiao , Zeng Hong , Feng Ru
Published: October 2019

Background

Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma (NHL). Although international prognostic index (IPI) has been widely used in DLBCL to evaluate the prognosis, there is need to find new biomarkers to predict outcome. Tumor microenvironment has been reported to be associated with both metastatic spread and diminished in some tumors. Tumor-associated macrophages (TAMs) and nature-killer (NK) cells play an important role in the tumor microenvironment. Therefore, we performed this study to investigate the prognostic implications of TAMs including M1 as well as M2 and NK cells in DLBCLs.

Materials and Methods

We reviewed 77 cases of DLBCL diagnosed between June 2004 and October 2012, who treated with rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP). Patients were divided into GCB and non-GCB subgroups by Hans’ algorithm. We performed immunohistochemical stains using CD68, CD163 and CD57 to mark M1 macrophages, M2 macrophages and NK cells on paraffin tissue section. The percentage of TAMs was recorded and graded as: 0 if less than 5%, 1 if 5-25%, and 2 if greater than 25% of the total cells present in tumor cells rich nodules were positive for CD68 or CD163. And counting the mean number of the CD57+ cells in 5 fields under high-power magnification (400 times): 0 if less than 5 positive cellsG1 if more than 5 positive cells.

Results

Of the 77 DLBCL patients treated with R-CHOP, there were 51 males (66.2%), 26 females (33.8%). The median age was 52 years (range, 16-81 years). The median follow-up was 38.1 months. The median percentage for CD68 was 12.8 %( range, 2.4–50.84), for CD163 was 12.3 %( range, 1.0–39.2). The median counts of CD57 cells were 5 (range, 1.0-78) per unit area. In univariate analysis, an increase in CD68-positive cells was related to improved EFS (grade 0 vs. grade 2, p = 0.036). By contrast, an increased number of CD163-positive cells were significantly poor associated with OS, (grade0 vs grade2, p = 0.033). Patients with elevated NK cells tended to have no effect on prognosis. In multivariate analysis, an increased CD68 -positive cells was an independent predictors of EFS (HR 0.467, 95%CI 0.236-0.922, P=0.028), an decreased CD163-positive cells and lower LDH were independent predictors for OS (HR 2.287, 95%CI 1.043-5.013, P=0.039).

Conclusion

These results suggest that M1 macrophages might predict favorable clinical outcome and M2 macrophages predict poor clinical outcome. However, the NK cells have no prognostic significance in DLBCL patients treated with RCHOP.

Disclosures

No relevant conflicts of interest to declare.
Immunohistochemistry in Staging Bone Marrow Biopsy Specimens: a Useful Adjunct for Morphological Diagnosis in Non Hodgkin’s Lymphoma
Authors: Arami Siamak , Svec Alexandr
Published: October 2019

Background: Morphological examination of bone marrow trephine biopsy represents a standard method for non-Hodgkin’s lymphomas (NHL) staging. Immunohistochemistry staining of bone marrow trephine specimens has been widely used in haematological malignancies due to its high applicability and sensitivity at diagnosis. However, the routine use of immunohistochemistry in the clinical settings when there is no obvious morphological (light microscopic) evidence of lymphoma in the bone marrow trephine, is not yet well established universally. We assessed the value of immunohistochemistry (by using a standard basic panel of anti-CD20 and anti-CD3 staining) in detecting involvement by NHL in routinely processed bone marrow trephine specimens with no obvious morphological involvement with lymphoma.

Methods: This study involved 56 randomly selected paraffin wax embedded, formalin fixed bone marrow trephine specimens between February 2011 and September 2013 from three teaching hospitals in Northeast, UK. 49 patients (87%) had B-cell NHL: diffuse large B cell lymphoma (DLBCL) 43% (n=24), follicular lymphoma (FL) 18% (n=10), marginal zone lymphoma (MZL) 9% (n=5), mantle cell lymphoma (MCL) 7% (n=4), lymphoplasmacytic lymphoma (LPL) 6% (n=3) and Burkitt’s lymphoma (BL) 6% (n=3). 7 cases (12%) had T/NK lymphomas. There was no obvious morphological evidence of bone marrow infiltration as all samples were reviewed by two examiners. All specimens were stained with the anti-CD20 and anti-CD3 antibodies.

Results: Concordant results were found in 50 samples (89%), as both investigations were reported negative. 6 of the 56 cases (11%) with no morphological evidence of involvement by NHL on routine stains, were positive on immunohistochemistry. Considering histology, discrepant results were noted more frequently in T/NK lymphomas (42%; 3 of 7 cases) comparing to B-cell NHLs (6%; 2 cases of DLBCL and 1 case of FL). In all six cases the lymphoid infiltrates had diffuse pattern.

Conclusions: Our results indicate that immunohistochemistry can detect a subgroup of NHL patients with bone marrow involvement beyond discriminatory level of conventional stains (Haematoxylin & Eosin and Giemsa), thereby contributing to accuracy of staging and treatment planning. Rational application of immunohistochemistry is a cost-effective & valuable method in routine investigation of staging bone marrow trephine biopsies.

Disclosures

No relevant conflicts of interest to declare.
The Glasgow Prognostic Score (GPS) As a Significant Predictor of Diffuse Large B Cell Lymphoma Treated with the R-CHOP in China
Authors: Li Xiaoyang , Zhang Yunxiang , Zhu Hongming , Hu Jiong , Li Junmin
Published: October 2019

Background: The Glasgow Prognostic Score (GPS), consisting of C-reactive protein (CRP) and albumin (Alb), is clinically useful for tumor behavior and shows significant prognostic value in several types of tumors. Several studies showed that GPS could be a predictive marker for advanced solid tumor, such as colorectal cancer, gastric cancer, etc. Now we performed a retrospective study to evaluate the relationship between GPS and prognosis of 160 patients with diffuse large B cell lymphoma (DLBCL) receiving R-CHOP chemotherapy between January 2001 and May 2010 at Shanghai Ruijin Hospital.

Methods: Before treatment, the CRP and albumin were both tested for base line. Patients with both elevated CRP level (>10 mg/L) and hypoalbuminemia (<35 g/L) were given a score of 2. Patients with only one of these two biochemical abnormalities were allocated a score of 1. Patients in whom neither of these abnormalities was presented received a score of 0. We also identified International Prognostic Index (IPI) score of these patients.

Results: Patients were assorted into three groups based on GPS at diagnosis. Survival curves were performed using the Kaplan-Meier method. Results showed that the overall survival (OS) and progress-free survival (PFS) rates for all patients were 78.1% and 68.1%, respectively. Analyzed by log-rank test, there was significant difference between GPS 1, GPS 2 and GPS 3 for OS (90.8% vs. 76.6% vs. 38.5%, p<0.001) and PFS (86.2% vs. 66.0% vs. 15.4%, p<0.001) (Fig 1a and b). In addition, the IPI, as a well-accepted prognostic system, though showed significant difference between lower risk group (IPI 0-2) and higher risk group (IPI 3--5) for OS (89.1% vs. 54.0%, p<0.01), failed to identify patients with poor prognosis in the lower risk group, since the low risk (IPI=0-1) and low-intermediate risk groups (IPI=2) appeared closely overlapping (p>0.05). While combining with the IPI model, GPS can have powerful prognostic value in the lower-risk (IPI=0-2) DLBCL patients, since patients with GPS score=0 had better OS (P<0.001) and PFS (P<0.001) than those with GPS score=1--2(Fig 2a and b).

Conclusion: The GPS was a reliable and independent predictor of OS and PFS in patients with DLBCL treated with R-CHOP regimen. GPS combining with the IPI score may offer a more integrated prognostic system in the rituximab era of DLBCL. And inflammatory response might play an important role in the progression and survival of patients with DLBCL.

Figure 1 Figure 1.

Figure 2 Figure 2.

Disclosures

No relevant conflicts of interest to declare.
Predictive and Prognostic Significance of Immunohistochemical Markers and Blood Absolute Lymphocyte Count in Diffuse Large B-Cell Lymphoma in Rituximab Era: A Polish Lymphoma Research Group (PLRG) Multicenter Retrospective Analysis
Authors: Czyz Anna , Joks Monika , Drozd-Sokolowska Joanna E , Knopinska-Posluszny Wanda , Zawartko Magdalena , Mizera-Nyczak Ewa , Dytfeld Dominik , Zdziarska Barbara , Jedrzejczak Wiktor-Wieslaw , Komarnicki Mieczyslaw
Published: October 2019

This report is a retrospective analysis of patients with newly diagnosed diffuse large B-cell lymphoma (DLBCL) who were treated with R-CHOP-21. The aim of the study was to determine the impact of peripheral blood absolute lymphocyte count (ALC), Ki67 proliferation index and immunohistochemical expression of Bcl-2 and Bcl-6 at diagnosis on complete response rate and survival.

The study group consisted of 161 patients with stage II-IV who were treated with six cycles of R-CHOP-21 between January 2004 and December 2013. With International Prognostic Index (IPI), 29% patients were low, 29% low-intermediate, 25% high-intermediate and 17% high risk. Bcl-2 and Bcl-6 expression was detected with immunohistochemical staining on paraffin sections in 83% and 77% of patients, respectively. Bcl-2 expression with high Ki67 proliferation index, greater than 80%, was found in 15% of patients. BcL-6 expression with Ki67 index of 80% or lower was detected in 52% of patients. Median ALC at diagnosis was 1.4 G/L (0.1-4.2). ALC below 1.0 G/L was revealed in 27% patients.

Results: The complete response rate to R-CHOP was 69%. In univariate logistic regression for complete response to immunochemotherapy, the variables that were correlated with lower response rate were IPI > 2 (p=0.016) and ALC < 1.0 G/L (p=0.014). In multivariate logistic regression, ALC < 1.0 G/L remained the only independent variable associated with failure to achieve complete response (OR 2.6; 95%CI 1.2-5.9; p=0.03). After the median follow-up time of 37 months (6-109), the 5-year progression free and overall survival (OS) estimated with Kaplan-Meier method were 67% (95% CI 56-76) and 66% (95% CI 55-75), respectively. In univariate analysis IPI (log-rank test, p=0.001), ALC (log-rank test, p=0.009) (fig.1), Bcl-2 expression with high Ki67 index (log-rank test, p=0.015) (fig.2) and Bcl-6 expression with low Ki67 index (log-rank test, p=0.05) were statistically significant for OS. In the multivariate analysis with Cox model, ALC at diagnosis (HR 2.9; 95%CI 1.1-7.3; p=0.024) and Bcl-2 expression with high Ki67 index (HR 2.8; 95%CI 1.1-7.5; p=0.039) remained significant for OS. IPI higher than 2 tended to impact negatively OS (HR 2.4; 95%CI 0.9-6.3; p=0.067).

In conclusion, our results indicate that low ALC at diagnosis is independent predictor of poor response to R-CHOP for patients with DLBCL. In addition, low ALC and Bcl-2 expression with high Ki67 index adversely influence patient survival in the rituximab era.

Figure 1 Figure 1. Figure 2 Figure 2.

Disclosures

No relevant conflicts of interest to declare.
Selective Inhibition of HDAC6 Decreases Viability of Cutaneous T-Cell Lymphoma and Improves Immune Recognition
Authors: Knox Tessa , Lienlaf Maritza , Perez-Villarroel Patricio , Lee Calvin , Sahakian Eva , Powers John J. , Cheng Fengdong , Canales Jorge , Marante Danay , Seto Ed , Sotomayor Eduardo M. , Villagra Alejandro , Horna Pedro
Published: October 2019

Cutaneous T-cell lymphoma (CTCL) includes a spectrum of non-Hodgkin’s T-cell lymphomas. While most patients with CTCL experience non-life threatening skin symptoms, those who do progress to later stages of the disease may develop serious complications. For patients with tumor stage or lymph node involvement there is a significant decline in treatment response & few treatment options are available. Current available therapeutic options include the use of histone deacetylase (HDAC) inhibitors as Romidepsin, Panobinostat and Vorinostat. However, their mechanism of action is still unknown. The role of histone deacetylases (HDACs) in cell biology, initially limited to their effects upon histones, now encompasses more complex regulatory functions that are dependent on their tissue expression, cellular compartment distribution, pathophysiological conditions and stage of cellular differentiation. Although major advances have been made in understanding the role of specific HDACs in cell proliferation and the survival of cancer cells, their individual participation in specific intracellular pathways is not completely understood. Here we present data demonstrating the changes in several immune-related pathways in two CTCL cell lines after exposure to both pan-HDAC inhibitors and selective HDAC6 inhibitors.

We compared the pharmacological effects of the pan-HDAC inhibitor LBH589 (Panbinostat), the class I selective Romidepsin, and the selective HDAC6 inhibitors such as TubastatinA and NexturstatB, on two CTCL cell lines (HuT78, HuT102). Our findings thus far demonstrate the following: First, we observed a marked inhibition of proliferation capacity of both cell lines when treated with either pan-HDACi or the more selective HDAC6 inhibitors. However, the selective HDAC6 inhibitors showed less cytotoxicity. Second, we observed important changes in the expression of the co-inhibitory molecules, such as PD-L1. Given our results, we conclude that the selective targeting of HDAC6 could recapitulate the anti-tumor effects of pan-HDAC inhibitors without having the non-target cytotoxic effects often encountered when using pan-HDAC inhibitors.

Disclosures

No relevant conflicts of interest to declare.