bluuebunny/crossref_metadata_2025 huggingface dataset:
New Gene Targets Detected By SNP Array Analysis In a Consecutive Series Of 215 Cases Of Adult Acute Lymphoblastic Leukemia
Authors: Safavi Setareh , Biloglav Andrea , Johansson Bertil , Paulsson Kajsa
Published: October 2019

Pediatric acute lymphoblastic leukemia (ALL) has a very favorable prognosis, effective treatment protocols and display a cure rate of more than 80%. However, adult ALL is associated with a much poorer prognosis; the long-term survival rate for adult cases is only a mere 30-40%, decreasing with higher age. For an improved prognosis and better treatment stratification in adult ALL, it is important to identify acquired genetic aberrations that could be linked to outcome. We present here the largest SNP array analysis to date, covering 5 million markers with a resolution of 10kb to investigate a consecutive series of 215 adult ALL cases, including samples obtained at diagnosis and relapse. We detected characteristic deletions, such as CDKN2A, PAX5, IKZF1, ETV6, RB1, and EBF1 genes.  In addition, the investigation uncovered several recurrent cryptic genetic events not previously implicated in adult ALL; the BCAT1, SERP2, RAB30, SRPR, ST3GAL4, ASS1, RASSF3, FUBP3, BCL11A, GAB1, LINGO2, TOX, and CXCR4 genes. Other frequent abnormalities included both partial and whole-chromosome uniparental isodisomies. Our findings provide insights into the leukemogenic process and may be clinically important in adult ALL.

Disclosures:

No relevant conflicts of interest to declare.
Minimal Residual Disease In Adult AML
Authors: Eldessouki Ihab A. , Khorshid Ola , Kandeel Eman , Lahloubi Nasr
Published: October 2019

Background

The achievement of complete hematologic remission (CR) is used as predictor for treatment response in patients with myeloid leukemia (AML).However <5% blasts in the bone marrow does not reflect the presence of tumor burden precisely. Minimal residual disease (MRD) in the first complete remission (CR1) may play a critical rule in assessment of treatment response and prediction of subsequent relapse.

Patients and Methods

Leukemia associated immunophenotyping (LAIP) for 73 patients with denovo AML monitored at diagnosis , day 14 and day28 post-induction by multiparametric flow cytometry (MFC).

Results

CR achieved in 60(82%) patients and 13(18%) patients did not. Among the 60(80%) patients who achieved CR 9 (15%) were MRD negative and 51(85%) were MRD positive at day14. Significant association between MRD detection and disease free survival (DFS) using 0.01% cut off value (P=.015). Day 28 post induction show highly significant association between MRD and DFS using 0.01% cut off value (P=0.001) as 38(63%) patients were MRD negative and (27%) were positive.

Significant association between MRD detection and overall survival (50 month) at day 14 and day 28 (P=0.02, P=0.001) respectively using cut off value 0.01%. MRD was positive in 63(86%) at day 14 and (37%) at day 28.

Conclusion

MRD detection at day28 and d14 at the end of induction in patients in CR may have a prognostic significance on clinical outcome and may thus be a useful marker for risk stratification.

Disclosures:

No relevant conflicts of interest to declare.
Stem Cell Programs Are Retained In Human Leukemic Lymphoblasts With Distinct Immunophenotypes
Authors: Hong Dengli
Published: October 2019

Leukemic lymphoblasts within different immunophenotypic populations possess stem cell Property. It remains largely unknown however whether self-renewal Program is retained from stem cells or endowed to Progenitors by leukemogenic molecules. We have addressed this issue in the context of TEL-AML1-associated acute lymphoblastic leukemia (ALL) by profiling a refined Program edited from genes essential for self-renewal of hematopoietic stem cells and B-cell development. Bioinformatic analysis shows that ALL populations are loosely clustered and all most close to the normal population that contains early lymphoid Progenitors, indicating that immunophenotypes do not reflect maturation stages in ALL and that self-renewal Program might be retained from stem cells. Results of assessing “first hit” function of TEL-AML1 in different populations of normal cells demonstrate the molecular model. Therefore, our studies reveal a leukemogenic scenario of human ALL that programs of stem cells are sustained in distinct fractions by leukemogenic mutations.

Disclosures:

No relevant conflicts of interest to declare.
EphA3 As a Target For Monoclonal Antibody Therapy For Acute Leukemia
Authors: Charmsaz Sara , Miller Kirrilee J , Day Bryan W , El-Ajeh Fares , Yarranton Geoff T , Bebbington Christopher R , Scott Andrew M , Lackmann Martin , Boyd Andrew W
Published: October 2019

Eph receptor tyrosine kinases interact with cell-surface ephrin ligands to direct cell growth, migration and cell positioning. High expression of EphA3 in a number of cancers has been linked to progression through facilitation of invasiveness and metastatic spread. EphA3 protein, which was originally described in leukemia, has also reported to be expressed in sarcomas, lung cancer, melanoma and glioblastoma. Using a Humaneered® derivative of the IIIA4 anti-EphA3 monoclonal antibody, KB004 (KaloBios Pharmaceuticals Inc), a clinical trial has commenced which targets the EphA3 protein in leukemia. The EphA3-specific monoclonal antibody, IIIA4, binds and activates human and mouse EphA3 with similar affinities, binding is followed by internalization of receptor-antibody complexes. We have shown high expression of EphA3 in the LK63 pre-B ALL and no expression of EphA3 in the Reh, a similar pre-B ALL cell line. We have examined the effect of the IIIA4 antibody in LK63 and Reh NOD/SCID xenograft models. In the LK63 xenograft model, administration of the IIIA4 antibody led to inhibition of tumour growth and decrease spread from bone marrow to the spleen and other organs and increased the latency of the disease, however no reduction in engraftment was observed in Reh xenograft model, suggesting that the effect was directed against the leukemic cells rather than the stromal and vascular elements. To further analyse the function of EphA3 expression, LK63 EphA3 knock down and EphA3-transfected Reh cells were tested in the xengraft model. Similar to LK63, antibody-treated Reh EphA3 xenografts showed reduction in the bone marrow engraftment and increased the latency of the disease. In contrast, LK63 EphA3 knock down xenografts showed minimal difference between treated and control group but notably showed a significant reduction in the splenic engraftment compared to the normal LK63 model. These results are consistent with IIIA4-mediated effects on the growth of EphA3-positive leukemic cells in the bone marrow and a secondary effect on the subsequent spread of the leukemic cells to extra-medullary sites and provides evidence that EphA3 is a functional therapeutic target in acute leukemias.

Disclosures:

No relevant conflicts of interest to declare.
Effects Of Metformin Addition To a Standard Chemotherapy Regimen For The Treatment Of Adult Acute Lymphoblastic Leukemia. An In Vitro and In Vivo study
Authors: Ramos Peñafiel Christian Omar , Carrillo Irma Olarte , Domínguez Jorge Zamora , Castellanos Sinco Humberto Baldemar , Fuller Etta Rozen , Peralta Enrique Miranda , Jaloma Juan Collazo , Tovar Adolfo Martínez
Published: October 2019

Antitumor effects of metformin are widely known. Unfortunately their use in the treatment of acute lymphoblastic leukemia is limited.

Study design

The study consisted of two stages

Phase 1: Cell assay adding metformin (40mM) assessing the viability and cell cycle in the cell line molt 4

Phase 2: Pilot study adding Metformin (850mg three time day) during the pre-treatment with steroids and the induction remission phase versus a control group (2:1 randomization)

Statistical analysis

Chi-square analysis was used to corroborate the hypothesis. The odds ratio was calculated for both, the absence of Good steroid response and refractory leukemia. Cox regression analysis and Kaplan-Meier curves were used for the survival analysis

Results

Celular assay. Metformin inhibited cell viability at 120hours reducing the percentage of cells in phase S.

Clinical assay. 151 patients were studied, 44 (29.1%) on Metformin arm, of these 59.1% ( n=26) archived a GSR compared with the control group (26.2%, n=28). A greater number of complete remission were presented in Metformin arm (81.8% versus 57.9%) shown to be a protective factor for GSR and complete remissions (odds ratio ; 0.2454 y 0.3062 respectively). In the Cox regression analysis, Metformin significantly impact in the global survival (p=0,012,95% IC) versus the other variables studied.

Conclusions

Metformin is useful both in vivo and in vitro treatment of adult acute lymphoblastic leukemia.

Disclosures:

No relevant conflicts of interest to declare.
Hyperthermia Enhances The Effect Of Magnetic Fe3O4nanoparticle Loaded With Daunorubicin On Leukemia and Drug Accumulation In Tumor In Vivo
Authors: Liu Ran , Wu Weiwei , Chen Baoan , Cheng Jian , Wang Jun , Zhang Jia , Bao Wen , Zhang Xiaoping , Zhang Dongsheng , Wang Xuemei
Published: October 2019

Multidrug resistance in cancer and adverse effects of chemotherapy are the major obstacles for cancer therapeutics. This study was aimed to investigate the efficiency of novel multifunctional Fe3O4 magnetic nanoparticles combined with chemotherapeutic agents and hyperthermia in order to reverse multidrug resistance and reduce side effects by concentrating chemotherapeutics on the target site in vivo leukemia models. K562 and K562/A02 xenograft tumor-bearing nude mice were randomly divided into 4 groups, a control group and the treatment groups were allocated to receive daunorubicin (DNR), Fe3O4 magnetic nanoparticles (Fe3O4-MNP), and Fe3O4 magnetic nanoparticles loaded with daunorubicin (DNR-Fe3O4-MNP). All groups were subjected to hyperthermia in an alternating magnetic field. Tumor volume was measured with a vernier caliper. Daunorubicin concentrations in plasma, tumor and non-tumor tissues were determined by high performance liquid chromatography (HPLC). Tumor and non-tumor tissues were stained with Hematoxylin and Eosin for histopathological and microscopic examination. Locations of Fe3O4 magnetic nanoparticles in tumor tissues were stained with Prussian blue for microscope examination. In Fe3O4-MNP and DNR-Fe3O4-MNP treated groups of both K562 and K562/A02 xenograft models, tumor temperature obviously increased and tumor volume became significantly smaller.  Apoptosis was observed in tumor cells treated with  DNR-Fe3O4-MNP groups.  In all DNR-Fe3O4-MNP groups of K562 and K562/A02 xenograft tumor models, the tumor inhibition rate, daunorubicin concentration  were higher level and  daunorubicin clearance in kidney also was delayed when compared with DNR alone treated group. Location of Fe3O4 magnetic nanoparticles with Prussian blue staining showed that Fe3O4 magnetic nanoparticles could be seen in tumor tissues. No obvious histopathological damage was observed in other non-tumor tissues.  Fe3O4 magnetic nanoparticles played an important role in increasing tumor temperature during hyperthermia, and can be delivered successfully to tumor site in an alternating magnetic field. Fe3O4 magnetic nanoparticles loaded with daunorubicin with hyperthermia had the strongest inhibitory effect on tumor and would be potential approach for improving the efficacy of chemotherapeutics, reducing side effects and reversing multidrug resistance in the treatment of leukemia.

Disclosures:

No relevant conflicts of interest to declare.
Impact Of Sex On Clinical Outcomes Of Mature B Cell Lymphoma In The Rituximab Era: A Multicenter, Retrospective Study
Authors: Nakamura Hiroyuki , Nagai Hirokazu , Watanabe Tomoyuki , Yano Takahiro , Uike Naokuni , Okamura Seiichi , Hanada Shuichi , Kawano Fumio , Sunami Kazutaka , Inoue Nobumasa , Yoshida Shin-ichiro , Shimomura Takeshi , Kitano Kiyoshi , Sawamura Morio , Horibe Keizo
Published: October 2019

Background

Rituximab has dramatically improved the clinical outcomes of mature B cell lymphoma. It has been reported that women show more favorable survival than men with rituximab-containing treatment. A multicenter, retrospective study was conducted to assess the role of sex in survival with rituximab treatment.

Patients and Methods

Patients with newly diagnosed mature B cell lymphoma treated at 20 National Hospital Organization hospitals in Japan from January 2000 to December 2004 were consecutively registered. Rituximab was approved in September 2002 for indolent B cell lymphoma and in September 2003 for aggressive B cell lymphoma in Japan. The patients were divided into two groups depending on whether they received induction therapy containing rituximab. The target population of this study was all mature B cell lymphoma patients who received first remission induction therapy containing rituximab. The patients treated without rituximab were used as controls. The endpoint was to compare 2-year progression-free survival (PFS) and overall survival (OS) between men and women. Survivals were assessed using the Kaplan-Meier method, and the groups were compared using the log-rank test.

Results

A total of 1126 patients received systemic chemotherapies during this study period. Of these, 348 (men 185, women 163), including 184 diffuse large B cell lymphomas (DLBCLs) and 111 follicular lymphomas (FLs), were treated by rituximab-containing regimens as front-line therapy. The 2-year PFS was better in women than in men (75.8% vs. 64.2%, p=0.048). This difference was not seen in the control group (men 396, women 382), which was treated by chemotherapeutic regimens without rituximab (48.7% vs. 50.6%, p=0.994). The 2-year OS was not statistically different between men and women (81.9% vs. 88.6%, p=0.115). When this population was broken down into DLBCL and FL, the women’s benefit in 2 year PFS was not statistically significant in both subtypes. Multivariate analysis both with forced entry and stepwise method could not show that sex was an independent prognostic factor in mature B cell lymphoma treated by rituximab containing induction regimen.

Conclusions

In mature B cell lymphoma, women would have better PFS than men when treated with rituximab containing therapy. These data suggest that the sex-based rituximab dose modification might be considered.

Disclosures:

No relevant conflicts of interest to declare.
Clinical and Laboratory Fectures Of T-Cell Prolymphocytic Leukemia In China
Authors: Zhang Yanru , Qi Junyuan , Qiu Lugui
Published: October 2019

Objective

To investigate the clinical and laboratory characteristics and survival of Chinese patients with T-cell prolymphocytic leukemia (T-PLL).

Method

Eleven patients with T-PLL were retrospectively analysed.

Result

There were nine males and two females, with the median age of 56.0(19-69) years old. All the patients, except for three, presented with leukocytosis. Marked leukocytosis was less frequent than that in the British series (75%) (P = 0.00). It was easy to see PB lymphocytosis with the median ALC of 17.22(0.58-148.83) ×109/L. Lymphadenopathy and splenomegaly were the most common physical finding. It was common that serum lactate dehydrogenase (LDH) and beta 2 microglobulin (β2-MG) were higher than normal. All cases were positive for CD2/ CD3/ CD5/TCRαβand negative for CD1a /HLA-DR and TdT, as for surface markers, including strong expression of CD7. By chromosome analyses, most cases (9/10)have normal chromosome .This rate is significantly higher than that of the British and American series (3% and 25%) (P =0.000, P =0.001). And 14q11 abnormality and trisomy 8q, which are common among Western cases, were not observed in any cases. With a median follow-up of 23 months, three cases died. Two year progress free survival (PFS) and overall survival (OS) were 53.3% and 50% , respectively. However, whether three cases with normal cytogenetics and indolent clinical courses could regard as “T-CLL” is needs to be investigated in future studies.

Conclusion

It was common that the increased lymphocyte count and lymphadenopathy as well as splenomegaly with T-PLL. And most cases have high level of blood LDH and β2-MG and normal chromosome karyotype. Of great interest is similar features are observed in cases in other East Asian countries such as Japan. Genome-wide analyses such as high-throughput mutation profiling, previously performed in other T-cell malignancies, will clarify the differences between Western cases and East cases in the future.

Disclosures:

No relevant conflicts of interest to declare.
Clinical Characteristics and Treatment Outcome Of CNS Involvement (Bing-Neel Syndrome) In Waldenstrom’s Macroglobulinemia
Authors: Kulkarni Tushar , Treon Steven Peter , Manning Robert , Xu Lian , Rinne Mikael , Lee Eudocia Quant , Ghobrial Irene M. , Norden Andrew , Kluk Michael J. , Nayak Lakshmi
Published: October 2019

Background

Waldenstrom’s Macroglobulinaemia (WM) is an IgM secreting lymphoplasmocytic lymphoma characterized by involvement mainly in the bone marrow, and occasionally in the lymph nodes and spleen. Central nervous system involvement of WM, known as Bing-Neel syndrome (BNS), is very uncommon and as such the clinical characteristics and treatment outcomes remain to be clarified.

Methods

We evaluated the incidence, clinical characteristics, and treatment outcome of BNS among patients with WM who were diagnosed at our Institution.

Results

We identified 13 patients with BNS from our database of 1,523 patients diagnosed with WM from 1999 -2013. The median age at diagnosis of BNS was 60 (range 51-75 years). The median time to development of BNS after WM diagnosis was 6.3 (range 0.3-11.9 years). Patients presented with a variety of neurological signs and symptoms including seizures, hearing loss, cognitive impairment, gait instability and lower extremity weakness. The diagnosis of BNS was confirmed by MRI in 11, and by examination of the CSF in 12 patients. CSF cytopathologic analysis demonstrated definitive evidence of malignant lymphoplasmacytic cells in 6 patients, and was suspicious for malignant cells in 2 patients. CSF flow cytometry was positive in 8, and a clonal immunoglobulin heavy chain gene rearrangement was identified in 6 patients. MYD88 L265P was identified in CSF sample of two patients in whom this examination was undertaken, and who demonstrated malignant disease by flow cytometry and cytological examination. Seven patients were treated with high-dose methotrexate (HD-MTX), 2 with intrathecal liposomal cytarabine, 1 with rituximab and intrathecal methotrexate, and 1 with rituximab and bendamustine. One patient was recommended treatment with HD-MTX and was lost to follow-up, 1 has been initiated on HD-MTX. Of 11 patients, 5 had a response in CSF and/or MRI, 4 had stable disease and 1 progressed following therapy. Of the responders, 2 patients had received HD-MTX, 2 intrathecal liposomal cytarabine and 1 rituximab-bendamustine. Four patients with stable disease had received HD-MTX. One patient who received intrathecal MTX developed treatment related “chemical meningitis”, and was subsequently treated with HD-MTX to which she had a CSF response. The median overall survival was not reached. Eleven patients are alive, 1 died and 1 was lost to follow-up.

Conclusions

BNS is an uncommon late complication of WM. MYD88 L265P may help in the diagnosis of BNS, and may represent a novel approach for targeted therapy of WM. HD-MTX is active in the treatment of BNS, though prospective studies are required to standardize treatment and improve outcomes.

Disclosures:

No relevant conflicts of interest to declare.
Bendamustine In Combination With Rituximab As First-Line Treatment For Indolent Non-Hodgkin Lymphoma: Retrospective Analysis Of An Spanish Registry
Authors: Solorzano Silvia , Martinez-Chamorro Carmen , Panizo Carlos , Quero Cristina , Deben Guillermo , Paz Jose , Batlle Ana , Serrano Alfons , Muentes Zayda , Gutierrez Antonio , Tomas Jose Francisco
Published: October 2019

Introduction

Indolent lymphomas represent 40% of all subtypes of non-Hodking's lymphoma, of which follicular lymphoma (FL) is the most frequent. Bendamustine is a dual alkylating agent with demonstrated high efficacy and low toxicity profile in reported clinical trials. We present the preliminary results from the experience of Spanish compassionate use registry of this agent as first-line treatment for indolent lymphoma.

Methods

Retrospective multicenter analysis of patients with indolent non-Hodgkin lymphomas (iNHL) treated as frontline with Bendamustine plus Rituximab (BR) in compassionate use. Clinical efficacy was evaluated according to Cheson criteria (2007) and toxicity according to CTCAE v3.0 scale. This study has been approved by local ethical committees.

Results

Patients’ characteristics: There are 96 patients registered (9 centers), with the following diagnoses: FL: 62 (64.5%), marginal zone 24 (25%), Waldenström macroglobulinemia 7 (7.3%) and mantle NHL 3 (3.1%). The main clinical features of the series are: 45% males, median age 64 years (range 36-84), 87.1% ECOG≤ 1, 63% Ann Arbor stage IV, 50.5% high risk FLIPI and 43.7% CIRS ≥ 4. Extranodal involvement was present in 79.1% of the patients, bone marrow  involvement in 52% and 11 patients (11.9%) had bulky disease. Treatment consisted in 6 cycles of BR (B-90 mg/m2 D1-2, R-375mg/m2 D1) in 95% patients. Median number of cycles administrated was 6 (range 1-8). G-CSF support was administered in 16.1% of cycles. Response and Safety: Overall response rate was 95%, with 65.5 % CR, 13.1% uCR and 16.4% PR in the 61 evaluable patients. Progression was documented in 4.9% of patients. Three exitus ocurred due to aspergillosis, progression and other not related with LNH. Median follow-up period was 14 months (3-47). In general, treatment was well tolerated; over 461 cycles registered, the most common adverse event was hematological toxicity with grade 3-4 neutropenia in 10.4%, grade 3-4 leucocitopenia in 6.9% and grade 3-4 anemia in 1.9% of the cycles. Other toxicities included all grades infections in 3.2% of patients, gastrointestinal in 3.4%, asthenia in 3.2%, chills in 1.1%, and mucositis 0.4%. Only 9 hospitalizations due to febrile neutropenia were reported.

Conclusion

Bendamustine plus rituximab was an effective and well tolerated regimen for newly diagnosed patients with indolent NHL.

Disclosures:

No relevant conflicts of interest to declare.
Risk Factors For Radiation Pneumonitis In Patients With Lymphoma Treated With Chemotherapy and Photon Or Proton Radiotherapy
Authors: Mesina Anna , Grover Surbhi , Mesina Carmen , Nasta Sunita D. , Svoboda Jakub , Plastaras John P.
Published: October 2019

Introduction

As volume-based radiotherapy planning has become a more standard part of combined modality therapy for lymphomas, dose-volume based constraints for organs-at-risk are needed for treatment planning. Conformal techniques such as intensity modulated radiation therapy and proton radiotherapy may achieve lower doses to certain organs like the heart, but may result in higher doses to other tissues like the lungs and breasts. We sought to define the dosimetric risk factors that are associated with development of radiation pneumonitis (RP).

Methods

This is a single-institution retrospective analysis of patients with thoracic lymphomas treated with combined modality therapy between 1999 and 2013 who had at least 4 months of follow-up after radiotherapy. Univariate analyses (UVA) were performed using Fisher exact and Wilcoxon rank-sum tests.

Results

Of the 89 patients analyzed, 13 (14.6%) were diagnosed with RP (at least Grade 1, Common Toxicity Criteria v4.0). Patients were predominantly female (62%) and never smoked (67%). Diagnoses were grouped as Hodgkin lymphoma (62.9%) or non-Hodgkin lymphoma (37.1%), and 18.5% were also treated with autologous stem cell transplants. UVA showed that RP was more commonly associated with a diagnosis of non-Hodgkin lymphoma (38%) than Hodgkin lymphoma (8%), despite exposure to bleomycin in the majority of patients with Hodgkin lymphoma. Higher lung doses were significantly associated with RP using multiple lung dose-volume parameters: mean lung dose (12.3 vs. 16.7 Gy, p=0.002), volume of lung receiving 20 Gy (27.3% vs. 39.1%, p=0.0009), and volume of lung receiving 5 Gy (51% vs. 66.8%, p=0.004). The majority (67%) of patients who developed RP had mean lung doses of over 15 Gy, whereas only 24% of those who did not develop RP had mean lung doses above 15 Gy, see Figure. Heart dosimetric parameters were also significantly associated with RP, including mean heart dose (13.3 vs. 21.5 Gy, p=0.004), volume of heart receiving 20 Gy (25.4% vs. 48.3%, p=0.003), and volume of heart receiving 5 Gy (57.6% vs. 81.1%, p=0.04). There were not enough events to determine if heart and lung parameters were independently associated with RP, but they were strongly correlated (R=0.75).

Gender, smoking history, and autologous transplant were not significantly associated with RP. None of the 13 patients treated with proton radiotherapy developed RP. In general, patients treated with proton radiotherapy had lower mean heart doses (9.4 vs. 15.4 Gy) and mean lung doses (9.6 Gy vs. 13.5 Gy).

Conclusions

Higher doses to lung and heart are associated with increased risk of RP, and doses to these critical structures should be considered carefully during volume-based consolidative radiotherapy using advanced techniques.

Disclosures:

No relevant conflicts of interest to declare.
Dose Of Vincristine didn’t Predict Outcome Of Treatment For Elderly DLBCL Patients Introductions
Authors: Miyata Yasuhiko , Nakamura Hiroyuki , Yamamoto Hideyuki , Kojima Yuki , Iida Hiroatsu , Nagai Hirokazu
Published: October 2019

Introductions

Diffuse large B-cell lymphoma (DLBCL) is one of the aggressive type of non-Hodgkin lymphoma (NHL) and is known to have a good drug-sensitivity and prognosis. More than 40% of patients with DLBCL are older than 70 years old at diagnosis, and age is known as a poor prognostic factor. However, dose modification is frequently necessary due to their comorbidities and organ function decline, and this leads to poor outcome of their treatment. Therefore, it is important to optimize treatment strategy and chemotherapy for the elderly with DLBCL to improve their outcome.

Aims

The aim of this study is to unveil underlying problems in the treatment against elderly patients with DLBCL and develop a new therapeutic approach to overcome them.

Methods

The medical records of all new elderly patients (70 years old< ) with DLBCL between 2003 April and 2011 October at Nagoya Medical Center were reviewed for a retrospective analysis , to identify characteristics and treatment response of those patients. Survival of patients intensively treated (82 pts) was evaluated using Kaplan-Meier method, and analyzed using Log-rank test.

Results

We experienced new 93 patients aged over 70 years (41.9 % of all DLBCL cases). Median observation period of was 52.7 months. R-CHOP and R-CHOP-like regimes were administrated into 78 and 4 patients respectively. Four patients treated without rituximab due to patients’ decision. CR was obtained in 70 patients (CR rate 85.3%), and 3-yr OS and PFS were 72.2% and 72.1% respectively. Eleven patients died during induction chemotherapy or didn’t obtain CR and eventually died. The median doses of ADR, CPM, and VCR were 69.51±15.2%, 70.23±20.0% and 59.1±28.1% of original regimen, respectively.% of VCR to original CHOP regimen was significant different between age 70-79 and age more than 80 (p<0.0001), but OS by % of actual dose VCR to original CHOP was not different (50%< vs. ≧50%, p=0.62), suggesting that dose of VCR didn’t predict outcome of treatment. It might suggest that dose of VCR could be reduced with keeping treatment outcome to improve QOL of survivors who suffer from peripheral neuropathy.

Disclosures:

No relevant conflicts of interest to declare.
Up Regulation Of Peroxiredoxin 2 Which Is Correlated With Disappearance Of Philadelphia Chromosome In CML Cells Might Have Therapeutic Applications During Imatinib Therapy
Authors: Yoo Eun-Sun , Mun Yeung-Chul , Ahn Jee-Young , Huh Jung-Won , Lee Kyoung Eun , Nam Eunmi , Lee Soon-Nam , Seong Chu-Myong
Published: October 2019

Background

BCR/ABL oncogenic tyrosine kinase is known to induce high levels of intracellular ROS which may further induce genomic instability with malignant transformation and even imatinib (IM) resistance. There are several lines of evidence that the production of reactive oxygen species (ROS) and the expression and activity of antioxidant enzymes make up the primary redox control of leukemia cells and cell survival. Therapies targeting the redox environment such as over-expression of antioxidants or antioxidant treatment, could inhibit tumor cell growth even resistant cells. In this study, we investigated the change of peroxiredoxin (Prx), which have a function of protection against oxidative stress, and survival of BCR-ABL positive cells during IM therapy.

Methods

BCR-ABL1 positive cell lines were cultured using 20% IMDM medium. We evaluated cell growth by MTT assay, and also evaluated BCR/ABL expression by western blot analysis. We also evaluated changes of intracellular ROS level and antioxidant enzymes, by immunoblot assay. siRNAs against Prx2 (100-200nM) were transfected into K562 cells using HiPerFect transfection Reagent (Qiagen) according to manufacturers’ protocol. During siRNA transfection, 1uM of imatinib was treated for 48hours. 72 hours after siRNA transfection, expression of BCR/ABL and Prx2 were detected by western blot assay and proliferation was determined by MTT assay method. Intracellular ROS was measured by flow cytometry after 10uM DCF-DA staining for 20mins.

Results

During IM treatment, we observed the significant decrease of cell viability and decrease of intracellular ROS level in BCR-ABL1 positive cell lines. These finding were well correlated with eradication of BCR/ABL oncogene by western blot results. The expression levels of Prx2 was restored with IM treatment and restoration of redox enzymes is correlated with reduction of BCR-ABL protein in Philadelphia positive cells. siRNAs transfected cells were not decreased cell survival even though eradication of BCR/ABL oncogene during IM treatment.

Conclusion

Significant change of ROS and Prx2 levels according to IM treatment are closely correlated with bcr/abl kinase level in both BCR-ABL1 positive cell lines. This finding was correlated with restoration of the level of PRX2. Those results has been implicated the possibility of therapeutic application by modulation redox system. Strategy of over-expression of Prx2 might be able to control sensitivity of the BCR-ABL1 positive cells to drug-induced apoptosis. Understanding the molecular mechanisms of changes on ROS and redox enzymes may be potential tools to develop more potent new drugs in Ph+ CML or Ph+ ALL patients especially for IM resistant.

Disclosures:

No relevant conflicts of interest to declare.
Hypoxia Regulates Eph/Ephrin-Facilitated Adhesion and Proliferation Of Chronic Myeloid Leukemia Cells
Authors: Zhou Xuan , Xiaoli Liu , Xu Na , Xiao Yajuan , Yi Zhengshan , Gao Guanlun , Zhang Jinfang , Huang Bintao , Du Qingfeng
Published: October 2019

Background and Objective

Recently, hypoxia has been demonstrated to be correlated with metastasis. The cellular response to oxygen deprivation involves a series of metabolic and biosynthetic events associated with adaptation to a hypoxic environment. Meanwhile, eph receptors and their ligands (ephrins) have also been proved to play a very important role in regulating the migrationand proliferationof leukemia cell in our early study. Therefore, we performed the experiment to address the hypothesis that hypoxia might be an important mediator linking Ephs/ephrins expression to chronic myeloid leukemia cell adhesion and proliferation.

Methods

K562 cells were cultured in a hypoxia environment with different density of oxygen (1%, 5%, 20%). The expression of HIF-1α, EphA5, EphB4, EfnA1, EfnB2 and signaling pathway proteins like p-FAK, p-Src, p-Paxilin, p-p130, p-cofilin, p-Akt, cyclin D3, cyclin E, p-Rb of K562 cells in different hypoxia environment were determined by real-time PCR and Immunoblot analysis. The K562 cells adhesion to endothelial cells and bone marrow stromal cells were tested with the adhesion assay kit. K562 cell proliferation, apoptosis and cell cycle were also detected with immunofluorescence analyses. After knocked down HIF-1α gene in K562 cells, the related protein levels and adhesion ratio were determined by the same methods.

Results

The results showed that the mRNA levels of HIF-1α were enhanced after 10 h of hypoxic exposure (1% O2), compared with normal exposure (P<0.05), as well as the mRNA levels of EphA5, EphB4, EfnA1, EfnB2 and related signaling pathway proteins were enhanced after 24h of hypoxic exposure (P<0.05). The protein levels of HIF-1α, EphA5, EphB4, EfnA1, EfnB2 and signaling pathway proteins like p-FAK, p-Src, p-Paxilin, p-p130, p-cofilin, p-Akt, cyclin D3, cyclin E, p-Rb also increased after 24h of hypoxic exposure (1% O2). After knocked down HIF-1α, the expression of Eph/ephrin didn’t enhance after 72h of hypoxic exposure(1% O2) ,which proved HIF-1α regulate Eph/ephrin expression. The adhension to endothelial cells was signiﬁcantly up-regulated (1.6 to 2-fold, P<0.05) after 36 h of hypoxic exposure (1% O2), the apoptosis cells down-regulated (1.3 to 1.5 fold, P<0.05).

Conclusions

We concluded that Eph receptors and ephrins were closely correlated with chronic myeloid leukemia cell adhesion and proliferation, and this process was regulated by hypoxia. Our findings shed new light on the regulation of this intriguing receptor/ligand family and present evidence for the role of Ephs/ephrins in chronic myeloid leukemia cells.

Disclosures:

No relevant conflicts of interest to declare.
Effective Targeting Of Chronic Myeloid Leukemia Initiating Activity With The Combination Of Arsenic Trioxide and Interferon Alpha
Authors: El Eit Rabab , Ahmad Iskandarani , Saliba Jessica , Jabbour Mark , Mahfouz Rami , Bitar Nizar , ElAyoubi Hanadi , Zaatari Ghazi , Mahon Francois-Xavier , de Thé Hugues , Bazarbachi Ali , Nasr Rihab
Published: October 2019

Imatinib is the standard of care in the treatment of Chronic Myeloid Leukemia (CML) patients. However, imatinib is not curative since most patients who discontinue therapy will relapse, stressing the need for novel therapies that target leukemia initiating cells (LIC). Arsenic trioxide induces complete remission in patients with acute promyelocytic leukemia (APL). We previously demonstrated that, through full degradation of PML-RARA, the combination of arsenic and retinoic acid eradicates APL LICs and cures the disease. Interferon alpha (IFN) induces hematologic and cytogenetic remissions and interestingly, improved outcome was reported with the combination of interferon and imatinib. We investigated the effects of arsenic and IFN on human CML cell lines or primary cells and the bone marrow retroviral transduction/transplantation murine CML model. In vitro, the combination of arsenic and IFN inhibited proliferation and activated apoptosis. Importantly, arsenic and IFN synergistically reduced the clonogenic activity of primary bone marrow cells derived from CML patients. Moreover, in a murine transplantation model of CML, we show that arsenic/IFN prolonged survival of primary leukemic mice and sharply diminished transplantation of CML cells in secondary recipients, pointing to exhaustion of CML LICs. Our results also suggest that IFN/arsenic may clear LICs independently from oncoprotein catabolism. Interestingly, arsenic/IFN effect on CML LIC activity was significantly superior to that of imatinib.These studies plea for a clinical exploration of this combination, knowing that IFN and arsenic have both shown clinical activity in CML, alone or in combination with imatinib.

Disclosures:

No relevant conflicts of interest to declare.
A Translocation (9;22)(p24;q11.2) In a Patient With CML
Authors: Abreu Daniele Costa , Castilho, Bachelor Ana Paula , Niewiadonski, Bachelor Vivian Dionísio , Drummond Mauricio , Gaburo Nelson
Published: October 2019

Introduction

In January 2013 was received in our lab service a bone marrow sample for cytogenetic analysis. The 61 years old female patient presents an elevated white blood cell count (118,000 x10³/mm³) and clinical diagnosis as Chronic Myeloid Leukemia (CML). According the medical information the treatment began with hydroxyurea 3g daily and allopurinol 300mg daily.

Methods

We proceeded with cytogenetic examination of the patient’s bone marrow aspirate by conventional G-banding analysis performed on unstimulated short-term cultures (24 hrs). FISH for BCR/ABL translocation was tested using a dual fusion dual color probe. Because of the sample stability we were unable to performed RT-PCR test.

Results

Chromosome analysis showed the translocation (9;22)(p24;q11.2) as a sole abnormality in 100% (20/20) of analyzed metaphases. Chronic myeloid leukemia presents as a specific chromosomal abnormality the Philadelphia chromosome, t(9;22)(q34;q11) which is different from the results obtained where the region of translocation of chromosome 9 was p24 instead of the classic q34. This result suggests it is BCR/JACK2 translocation. The FISH analysis showed the presence of a complex Ph chromosome: ABL con BCRx1 (one fusion) and BCRx2;ABLx2.

Conclusion

The patient took imatinib without answer. She is still in clinical monitoring with persistent hyperleucocytosis and the treatment is following with hydroxyurea 500mg daily and Interferon 5000 UI three times a week. Further molecular and cytogenetic tests will be performed in a second sample to contribute with evaluation of disease progression and monitoring treatment response.

Disclosures:

No relevant conflicts of interest to declare.
A Decade Of Imatinib As First-Line Treatment Of Chronic Myeloid Leukemia: A Single - Latin American Institution Experience
Authors: Garcia Alvaro Cabrera , Delgado Carolina Balderas , Kassack Ipiña Juan Julio , Ramos Peñafiel Christian Omar , Jaloma Juan Collazo , Tovar Adolfo Martínez , Gallardo Hermelinda , Castellanos Sinco Humberto Baldemar
Published: October 2019

Targeted therapy commenced against Chronic Myeloid Leukemia (CML) with the development of small-molecule tyrosine kinase inhibitors (TKIs), since its approval in 2001 as first-line therapy, imatinib has been effective in achieving high response rates and improving the prognosis. However, the excellent results of large clinical trials are not entirely reproducible in the “real world”, we want to share our experience of a decade of use as front-line treatment.

A total of 160 patients with previously untreated CML since 2002 were reviewed: 90% chronic phase (CP), 7% accelerated phase (AP), and 3% blast phase (BP). All were treated with standard-dose imatinib. In CP, overall survival (OS) at 75 months was 82% and progression-free survival 66%(PFS). Survival was worse in those over 60 years (P= < .001). 40% achieved and maintained Complete Cytogenetic Response (CCyR), 19 patients progressed to BP. 23.4% had some degree of hematologic toxicity that required a temporary suspension or reduction of the initial dose. 26.8% of patients with chronic phase treated at our institution were in the high EUTOS score. In this population, the EUTOS score was not predictive for outcome. Our study showed that imatinib is effective, and well tolerated by our patients and remains as a good strategy used as first-line therapy for patients with CML.

Disclosures:

No relevant conflicts of interest to declare.
The Depth Of ‘Complete’ Molecular Response Predicts Molecular Relapse In Chronic Myeloid Leukaemia Patients On Tyrosine Kinase Inhibitor Therapy
Authors: Thomas Heledd , Apperley Jane F , Szydlo Richard M , Gerrard Gareth , Marin David , Milojkovic Dragana , Foroni Letizia
Published: October 2019

Background

There is currently no universal definition of complete molecular response (CMR) in chronic myeloid leukaemia (CML). This has implications for studies of TKI withdrawal as CMR is the main criterion for entry into these trials. We aimed to assess the proportion of patients in our practice who achieved CMR by a variety of definitions. We hoped to identify a definition that was predictive of sustained CMR and could therefore be recommended to predict those patients who might be able to stop treatment in the longer term.

Methods

We conducted a retrospective analysis of a comprehensive CML database of serial RT-qPCR results of 215 patients achieving deep molecular responses on TKI therapy. The least stringent definition of CMR was defined as BCR-ABL1 transcripts <11 and ABL1 control value (CV)>10,000. The depth of CMR was categorised according to transcript number and CV. Probability of molecular relapse according to depth of CMR at 2 and 5 years from initial CMR (ICMR) was estimated using the Kaplan Meier method.

Results

Patients with 6-10 transcripts (any CV) were most likely to lose CMR by 2 years (86.4%) while patients with 0 transcripts (any CV) were least likely to do so (7.6%). There was no difference in the probability of molecular relapse between patients with 1-5 transcripts (CV>32,000) and those with 0 transcripts (any CV) (RR at 2 years=1 p=0.945, RR at 5 years=3.7 p = 0.114).

Conclusion

Depth of CMR predicts molecular relapse at 2 and 5 years from ICMR. CMR4.5 (<6 transcripts) is the least stringent response that is required to sustain CMR. Patients with <6 transcripts (CV>32,000) are equally as likely to sustain CMR as patients with 0 transcripts up to 5 years from ICMR. These patients may be suitable for a stopping trial.

Disclosures:

Apperley: Ariad: Honoraria; Bristol Myers Squibb: Honoraria; Novartis: Honoraria, Research Funding; Pfizer: Honoraria. Milojkovic:BMS: Honoraria; Pfizer: Honoraria; Ariad: Honoraria; Novartis: Honoraria.
Prognostic Factors Associated To Achievement Of Complete Or Partial Response In MDS Patients Treated With Azacitidine Outside Clinical Trials
Authors: Breccia Massimo , Voso Maria Teresa , Maurillo Luca , Niscola Pasquale , Autore Francesco , Aloe Spiriti Maria Antonietta , Buccisano Francesco , Tatarelli Caterina , Tendas Andrea , Fenu Susanna , Latagliata Roberto , Alimena Giuliana
Published: October 2019

Azacitidine is able to induce responses and to prolong survival (OS) in intermediate/high risk MDS subgroups or in CMML: in the AZA001 trial, median OS was significantly longer for azacitidine patients compared with the combined conventional care regimen (CCR) group. We retrospectively assessed prognostic factors associated to the achievement of these responses in a large series of MDS patients treated outside clinical trials. Overall, we recruited for this study, 166 primary or secondary MDS and 21 CMML diagnosed and treated with azacitidine in 6 different hematologic units. Patients were recruited consecutively, without any criteria of exclusion. All patients received azacitidine with a schedule of 5+2+2 or of 7 consecutive days every 28 days until progression or unacceptable toxicity. Clinical parameters (age, sex, WHO classification, IPSS) and baseline cytogenetic evaluation were retrospectively collected. Of 166 MDS patients recruited, 103 were males and 63 females; median age was 69.5 years (range 49-89). A median of 8 cycles was performed (range 1-60). According to IPSS stratification there were 29 intermediate-1 risk patients, 118 intermediate-2 patients and 15 high–risk patients (7 patients not determined). According to WHO classification, 37 patients were diagnosed as having RAEB-1, 101 patients as RAEB-2, 28 patients as RCMD. Of 21 CMML patients recruited, 19 were males and 2 females; median age was 67.5 years (range 62-89). According to WHO classification, 14 patients were CMML-2. Overall, 41 patients (22%) achieved a CR/PR according to IWG 2009 criteria. Analysis of factors associated to the achievement of best responses revealed that pre-azacitidine WBC higher than 5 x 109/l (p=0.02) neutrophil count higher than 0.5 x 109/l (p=0.03) and platelet count higher than 150 x 109/l (p=0.01) were significantly associated to CR/PR achievement. We did not reveal significant differences, when compared to patients that did not achieve a response, in terms of sex, age, hemoglobin level at baseline, bone marrow blast count, cytogenetic profile, IPSS stratification. In conclusions, from our results, based on a large series of patients treated outside clinical trials, it seems that, independently from cytogenetic or hematological subtype, only clinical parameters at baseline could influence the achievement of a good response with azacitidine.

Disclosures:

No relevant conflicts of interest to declare.
Long-Term Outcome Of Higher-Risk MDS Patients Treated With Azacitidine: Single Centre Experience
Authors: Ricco Alessandra , Sgherza Nicola , Rossi Antonella Russo , Carluccio Paola , Franco Adele , Pasciolla Crescenza , Casieri Paola , Specchia Giorgina
Published: October 2019

Background

The primary goals of treatment for patients with higher-risk MDS are to improve bone marrow function, suppress AML transformation and prolong survival. The international phase 3 trial, AZA-001, established that azacitidine (AZA) significantly prolonged median overall survival and significantly delayed median time to AML transformation as compared with conventional care regimens. AZA was also associated with significant improvements in other clinically relevant outcomes, including reducing the need for transfusion, hospitalization and intravenous antimicrobials, and improving quality of life. Although several clinical trials have examined the safety of AZA, limited data are available about the long-term use of this drug.

Aims

We report a retrospective analysis of the outcome of 16 higher-risk MDS patients (M/F: 7/9) treated at our Institution with AZA, from June 2008 to June 2013, with a median follow-up of 33.5 months (range 24-60) after the start of AZA to determine the characteristics of this subset of long-term survivors.

Patients and Methods

Median age at diagnosis was 68 years (range 51-75) and PS was <2 in 13 cases (81.2%). Diagnosis according to the 2008 WHO Classification was refractory anemia with excess blasts (RAEB)-1 in 7 cases and RAEB-2 in 9 cases. Cytogenetic risk was low in 6 cases (37.5 %; all with normal karyotype) and high in 10 cases (62.5%; all with complex abnormalities). All patients received AZA administered subcutaneously at a dosage of 75 mg/m2 daily for 7 days, repeating the cycle every 28 days; median number of AZA cycles received was 25 (range 19-56).

Results

According to International Working Group 2006 criteria, 4 patients (25%) achieved complete response, 7 patients (44%) achieved partial response, and 5 (31%) stable disease. 13 patients (81%) are still alive and continuing to receive AZA. Two patients (12.5 %) died due to disease progression, one after 27 months and one after 29 months after the start of AZA. One patient died to infection disease (pneumonia). AZA treatment has lasted more than 3 years in 4 (25%) of the 16 patients. Peripheral cytopenias were the most common advent events for treatment.

Conclusions

Allogeneic stem cell transplantation remains the only potentially curative treatment for MDS but the vast majority of patients with MDS are ineligible for transplantation because of comorbidities, advanced age, or lack of a suitable donor. Study in a larger series of patients is warranted, but our experience shows that in the absence of intolerable toxicity or disease progression, continued AZA treatment seems to improve the outcome of higher-risk MDS patients who are not eligible for transplantation.

Disclosures:

No relevant conflicts of interest to declare.