bluuebunny/crossref_metadata_2025_split huggingface dataset:
Successful Peripheral Haematopoitic Autologous Stem Cells Transplants After More Than 5 Days of without Cryopreservation In Non Hodgkin Lymphoma: Asian Scenario
Authors: Jasuja Sandeep , Malhota Hemant , Kukar(Jasuja) Neetu , Gautam Dinesh , Sharma Shrikant , Sharma Rajesh 
Published: October 2019

Abstract
Abstract 4447

Background:
Methods to simplify the stem cell transplantation procedure are the need of the hour in developing countries due to constraints of affordability and the large number of eligible patient for transplantation. Unprocessed leukapheresed material is useful to restore hemopoiesis after high dose chemotherapy and, in addition to being significantly less expensive, it also avoids the toxicity of cryopreservation (DMSO).

Method:
Over a period of 18 months (January 2009 to June 2010) in a hospital setting, we prospectively performed auto transplants in 5 relapsed/resistant non-Hodgkin's lymphoma patients using non-cryopreserved (stored for 5–8 days at 4°C) and un-manipulated peripheral blood stem cells mobilized by GCSF (10mg/kg/day) and GMCSF (5mg/kg/day) using a 5–7 day conditioning regimen. Viability of stem cells was checked daily by trypen blue and the cell were reinfused cells when viability comes down to less then 80% (minimum 5 days) or on day 8 which ever was earlier. The median collection of CD34+ stem cells was 2.8 × 106/cu mm (range 2–4.8 × 10 6/cumm)

Result:
The median time to achieve > 0.5 × 109/l granulocytes was 14 days. The median time to achieve > 20 × 109/l platelets was 16 days. CR was achieved in 4 cases. A very good partial response was achieved in 1 case. The 100-day mortality was zero. One patient, who was not in CR, died from relapse of the lymphoma, 9 months post transplant. The overall median post-transplant survival for other patients has not been reached. The approximate cost of the each graft was US$6,000.

Conclusion:
From this small series, we conclude that a simplified method to autograft patients avoiding purging procedures and cryopreservation of the cells is feasible. This technique helps avoid toxicity of DMSO and significantly decreases the cost of autologous hematopoietic stem cell transplant, perhaps in the world, an important issue in developing countries.

Disclosures:
No relevant conflicts of interest to declare.
Rapid Reduction of Chronic Myeloid Leukemia Stem Cells After Treatment with Second-Generation BCR-ABL Kinase Inhibitors, Dasatinib and Nilotinib
Authors: Minami Yosuke , Abe Akihiro , Nomura Yuka , Minami Miho , Kuwatsuka Yachiyo , Kitamura Kunio , Hiraga Junji , Mizuno Shinichi , Yamamoto Kazuhito , Kajiguchi Tomohiro , Sawa Masashi , Inagaki Yuichiro , Yokozawa Toshiya , Aoki Etsuko , Ozeki Kazutaka , Watanabe Keisuke , Miyamura Koichi , Kiyoi Hitoshi , Naoe Tomoki 
Published: October 2019

Abstract
Abstract 4457
Chronic myeloid leukemia (CML) is effectively treated with imatinib (IM), however, several mathematical models and ex vivo-examinations suggested that IM-therapy does not eradicate BCR-ABL-positive hematopoietic stem cells (HSC). We prospectively (0, 3, 6 and 12 months after IM-therapy) investigated 16 newly diagnosed and 22 long-term followed CML-chronic phase (CP) cases using methods previously reported (Jamieson et al., N Engl J Med, 2004. and Abe et al., Int J Hematol, 2008) (Figure 1) with FACSAria™ and quantitative RT-PCR of BCR-ABL among each sorted population; total mononuclear cells, HSC/Thy-1+, HSC/Thy-1–, common myeloid progenitors (CMP), granulocyte macrophage progenitors (GMP) and megakaryocyte erythroid progenitors (MEP). In optimal responders to IM-therapy, BCR-ABL transcripts in the HSC populations (HSC/Thy-1+ and HSC/Thy-1–) tended to be more retentive than other populations while gradual reduction was observed during the first 12 months in all populations. And discrepancy of minimum residual diseases (MRD) between the HSC populations and other populations was larger in patients after longer IM-therapy. In evaluating properties of CML stem cells and other markers, we observed irrelevant distribution of side population (SP) and expressions of ABC transporters (ABCB1 and ABCG2) in comparison with CD34/38 expression. We also prospectively investigated BCR-ABL transcripts in each population of 23 IM-resistant or -intolerant CML-CP cases and one newly diagnosed CML-accelerated phase (AP) case during treatment with second-generation tyrosine kinase inhibitors (2nd TKIs), dasatinib or nilotinib. Treatment with each inhibitor induced more rapid reduction of BCR-ABL transcripts even in the HSC population (CD34+CD38–) during the first 6 months and there was no significant difference of MRD among each population in optimal responders to 2nd TKIs-therapy. In the stromal co-culturing system using primary cells and leukemic NOD/SCID/IL2rgnull (NOG) mice xenotransplanted with Ph+ leukemia cells, retention of quiescent slow-cycling (Hoechst 33342low/Pyronin Ylow) CD34+ population after IM-treatment were observed and cell death mechanisms after treatment with 2nd TKIs are also under investigation. These results imply that therapy with 2nd TKIs could be a promising approach for quick and efficient reduction of the CML stem cells and cure of disease.
Figure 1 Figure 1.

Disclosures:
Naoe: Kyowa-Kirin: Research Funding; Novartis: Research Funding; Bristol-Myers Squibb: Research Funding.
Regulation of c-Myc Expression through the Depletion of THAP11 and Fbw7 by Bcr-Abl Promotes CML Cell Proliferation
Authors: Yokota Daisuke , Nakamura Satoki , Takemura Tomonari , Nagata Yasuyuki , Hirano Isao , Ono Takaaki , Shigeno Kazuyuki , Fujisawa Shinya , Ohnishi Kazunori 
Published: October 2019

Abstract
Abstract 4463

[Background and Aims]
In various signaling pathways activated by Bcr-Abl, c-Myc promotes the Bcr-Abl transformed cell proliferation through activated Akt, Jak2 and NF-κB. We have previously shown that Bcr-Abl represses the expression of THAP11 and continuously expressed c-Myc, resulting in the proliferation of CML cells. However, the mechanism of c-Myc regulation is unclear in CML cell proliferation. In the present study, we investigated the mechanism of c-Myc transcription regulation in the Bcr-Abl+ progenitor cells derived from CML patients.

[Methods]
The cells used in this study were human CML cell lines, K562, Meg01 and SHG3 cells. Primary CML cells (ALDHhi cells) were obtained from the bone marrow of CML (CP) patients (n=12). Human normal ALDHhi cells were isolated from bone marrow of healthy volunteers after obtaining informed consents. For analysis of THAP11 mRNA expression, quantitative RT-PCR was performed in all cell lines treated with Abl kinase inhibitors (STI571, AMN107, and BMS354825). For cell survival analysis and the levels of c-Myc, Fbw7 and some CDKIs in CML cells, MTT assays, western blot and cell cycle analysis were performed in all cell lines transfected with THAP11 siRNA or cDNA. For colony analysis, the colonies of CFU-GEMM, CFU-GM, and BFU-E were counted in CML stem/progenitor cells transfected with THAP11 siRNA or cDNA, or treated with Abl kinase inhibitors.

[Results]
In CML cell lines, the expressions of THAP11 mRNA and protein were significantly increased by treatment with Abl kinase inhibitors or transfection with Bcr-Abl siRNA. In CML cells transfected with the THAP11 cDNA, it is shown that CML cell proliferation was inhibited, and the expression of c-Myc protein was decreased compared to the untransfected cells. In addition, the inhibition of Bcr-Abl by the Abl kinase inhibitors or depletion of Bcr-Abl induced the expression of Fbw7 in CML cells. In CML stem/progenitor cells (ALDHhi cells) obtained from patients with CML, the expression of THAP11 mRNA was suppressed, and the transfection with Bcr-Abl siRNA or treatment with Abl kinase inhibitors increased the expression of THAP11 and Fbw7. In CFU-GEMM, CFU-GM, and BFU-E, treatment with the Abl kinase inhibitors and depletion of Bcr-Abl induced the THAP11 and Fbw7 expression and reduced the c-Myc expression, and inhibited colony formation of Bcr-Abl+ progenitor cells.

[Conclusion]
In CML cells, we found that the depletion of THAP11 and Fbw7 expression. We showed that Bcr-Abl induced the expression of c-Myc by depletion of THAP11 and also inhibited the degradation of c-Myc by depletion of Fbw7. This study showed that Bcr-Abl promoted the CML cell proliferation through the THAP11 and Fbw7 mediated-c-Myc aberrant expression.

Disclosures:
No relevant conflicts of interest to declare.
Are CML Lymphoid Blast Crisis and Ph Positive ALL Genomically Indistinguishable?
Authors: Grace Colin , Nacheva Elisabeth P 
Published: October 2019

Abstract
Abstract 4464
Philadelphia positive malignant disorders are a clinically divergent group of hemoblastoses with a unique identifying feature, the BCR/ABL1 fusion gene, usually resulting from the chromosome rearrangement t(9;22)(q34;q11) or its variants, that leads to constitutive expression of an aberrant tyrosine kinase. These include chronic myeloid leukaemia (CML) and de novo acute leukaemia of both myeloid Ph(+)AML and lymphoid origin Ph(+)ALL. The latter two disorders are clinically aggressive and therapy challenging even in the era of the powerful tyrosine kinase inhibitors. CML is a multistage progressive disease, which if untreated, inevitably ends as fatal acute leukaemia, either myeloid or lymphoid. The latter is often thought to be indistinguishable from Ph(+)ALL, the most common type of ALL in adults. We have identified DNA sequences the imbalances of which appear to be significantly associated with the disease stage and lineage origin in CML and Ph(+)ALL samples. We used array CGH at a resolution of ~2kb to explore hot spot regions obtained from 102 patient samples comprising 92 CML and controls together with 10 Ph(+)ALL and show how Significance Analysis of Microarrays (1) can be used to identify differences in the genome profile of de novo Ph(+)ALL and lymphoid blast transformation of CML. We show that lymphoid blast crisis CML differs significantly from Ph(+)ALL not only due to the presence of 9p deletions but also due to genomic gains in other chromosomes. Furthermore we identify a sub group of Ph(+)ALL with a distinctive genomic profile. Having identified genome regions of potential interest, ranked in order of significance, out of the 100's of thousands of array results, it is then a challenge to design further experiments to evaluate their contribution to the biology of the BCR/ABL positive disease.
1 Tusher V, Tibshirani R, Chu G: Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci U S A 98:5116-5121 (2001).

Disclosures:
No relevant conflicts of interest to declare.
G2/M Arrest Sensitizes Chronic Myelogenous Leukemia Cells to TRAIL-Induced Apoptosis
Authors: Devlin David , Szegezdi Eva , Tanja Paavilainen , Orosz Orsolya , O'Dwyer Michael , Carty Michael , Samali Afshin , Hennequart Séverine Cruet 
Published: October 2019

Abstract
Abstract 4465
The death ligand, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receives great interest as it targets and kills cancerous cells, but not non-transformed cells. While it is in phase I/II clinical trials for a range of solid tumours, the generally low sensitivity of leukemia cells to TRAIL makes it a less attractive therapeutic for these cancers.
We found that doxorubicin and cytarabine, agents that induce DNA damage and impair cell cycle progression, can sensitize CML cells to TRAIL with CI<1 at Fa of ED25 and ED50 (based on median-effect method using the isobologram equation). Inhibition of the cell cycle checkpoint kinases Chk1/2 with UCN-01 did not influence TRAIL-induced apoptosis nor could it abolish the sensitizing effect of doxorubicin. Interestingly, inhibition of Ataxia Telangiectasia Mutated (ATM), a key DNA damage response kinase, with KU-55933 induced a G2/M arrest and enhanced TRAIL-induced apoptosis. Inhibition of ATM alone induced 22±3.1% apoptosis and increased TRAIL-induced apoptosis from 27.2±4.7% to 68±7.2%. Cell cycle analysis revealed that while the proportion of cells in the G0/G1 and S phases slightly increased, the proportion of the cells in the G2/M phase dropped by 31.6±3.2% (p<0.05) indicating that G2/M arrested cells were more sensitive to TRAIL than cells in G0/G1 and S phases. TRAIL-induced CML cell death was also synergistically enhanced by arresting the cells in G2/M using the microtubule disrupting drugs, nocodazole or colcemide. Cells were treated with a concentration of nocodazole or colcemide that induced above 90% G2/M arrest for 16 h (0.3 mM and 0.1 mg/ml, respectively) followed by treatment with 250 ng/ml of TRAIL for 24 h. Nocodazole, colcemide and TRAIL individually induced 19±3.7% 26.3±4.4% and 27.2±4.7% cell death, while combination of nocodazole or colcemide with TRAIL resulted 89±6.8% and 93±5.9% cell death, respectively. In summary, we found that induction of DNA damage sensitizes CML cells to TRAIL and that TRAIL-sensitivity of CML cells is cell cycle-dependent.

Disclosures:
O'Dwyer: Novartis: Honoraria.
Clinical Relevance of Vascular Endothelial Growth Factor Type A (VEGFA) and VEGF Receptor Type 2 (VEGFR2) Gene Polymorphism In Chronic Lymphocytic Leukemia
Authors: Gora-Tybor Joanna , Jamroziak Krzysztof , Szemraj Zofia , Szemraj Janusz , Lech-Maranda Ewa , Grzybowska-Izydorczyk Olga , Oszajca Katarzyna , Giannopoulos Krzysztof , Robak Tadeusz 
Published: October 2019

Abstract
Abstract 4467
Vascular endothelial growth factor type A (VEGFA) is a key regulator of angiogenesis and vascular permeability. Given that chronic lymphocytic leukemia (CLL) cells secrete VEGFA, express VEGFA receptors and circulate in blood with increased VEGFA level it is likely that VEGFA-mediated signaling may influence CLL clone survival. In this case-control study we verified the hypothesis that inherited differences in activities of VEGFA and its main receptor in CLL, vascular endothelial growth factor receptor type 2 (VEGFR2, also known as kinase insert domain-containing receptor/fetal liver kinase-1, Flk/1/KDR) impact predisposition to CLL or the course of the disease. Four functional single nucleotide polymorphisms (SNPs) including two SNPs in VEGFA gene, namely +405G>C (rs2010963) and +936C>T (rs3025039) and two SNPs in VEGFR2 gene, namely -271G>A and +1719A>T (rs1870377) were genotyped using PCR-based assays in 175 Caucasian CLL patients and 133 ethnically, geographically, age and sex matched controls. The minor allele frequencies in CLL patients were found as follows: 0.36 for +405C VEGFA allele, 0.17 for +936T VEGFA allele, 0.46 for -271 VEGFR2 allele and 0.35 for +1719A VEGFR2 allele. The genotype and allele frequencies of all tested SNPS in VEGFA and VEGFR2 genes did not differ significantly between cases and controls (p>0.05). No genotype or allele was significantly associated with important prognostic factors in CLL including clinical stage, IgVH mutational status, ZAP-70 expression and FISH cytogenetic abnormalities. Furthermore, the carriers of different VEGFA and VEGFR2 genetic variants had comparable time to CLL treatment initiation. In conclusion, the results of our study do not support major involvement of genetic polymorphisms in VEGFA mediated pathway in susceptibility to CLL or its outcome.

Disclosures:
Robak: GlaxoSmithKline: Consultancy, Honoraria, Research Funding; Genmab: Consultancy, Research Funding; Roche: Consultancy, Honoraria, Research Funding.
Sustained Complete Molecular Response (CMR) with Interferon-alpha Maintenance After Induction with Imatinib In Chronic Myeloid Leukemia (CML) Patients.
Authors: Carella A. M. , Catania G. , Pica and G. Beltrami G. 
Published: October 2019

Abstract
Abstract 4486
Cytotoxic lymphocytes (CTLs) which recognize distinct antigenic peptides on CML cells, contributed greatly to the cure after allografting. Previously, BCR-ABL specific peptides and the PR1 non-apeptide, were found to elicit a CML-specific CTL response (Molldrem, 1996/1997; Bocchia 1996, Clark 2001). PR1 is a peptide derived from myeloblastin (MBN), also known as proteinase 3. MBN is abundantly expressed in azurophil granules of normal myeloid cells and is substantially overexpressed in certain immature myeloid leukemia cells where it may be important for the maintenance of a leukemic phenotype. PR1-specific CTLs (PR1-CTLs) lyses and inhibits the proliferation of CML cells but not of normal myeloid precursors. The association between the emergence of PR1-CTLs and the response to IFN-a in vivo strongly implied that IFN-a can induce remissions via induction of a CML-specific PR1-CTLs response (Molldrem et al, 2000).
Recent data demonstrating a failure of IM to kill cell cycle-arrested CML precursors advice us to be attentive to arising escape mechanisms of even chronic phase CML; therefore IFN-a, given to CML patient with undetectable disease achieved with IM, might enable its discontinuation.
We have treated until now 8 CML patients who achieved stable CMR for at least one year after IM therapy. These patients discontinued IM and started IFN-a at the dose of 3–9 M/U/m2 weekly (median 6 M/U/m2). The median duration of IM therapy was 60 (range, 31–76) months before IM discontinuation. At a median follow-up of 16 months (range, 9–24 months) after discontinuation of IM, five patients still have an undetectable level of BCR-ABL transcript under IFN-a. Grade 2 intolerance (muscle pain, fever) to IFN-a developed in 5/8 patients. The details of patients characteristics will be discussed during the presentation. These preliminary results demonstrate that treatment with low dose IFN-a in CML patients with undetectable disease is feasible and this approach may become an attractive alternative to lifelong TKI therapy.

Disclosures:
No relevant conflicts of interest to declare.
Extensive Pleural and Pericardial Effusion In 4 CML Patients Treated with Dasatinib at 100 Mg or 50 Mg Once Daily.
Authors: Krauth Maria-Theresa , Herndlhofer Susanne , Schmook Maria-Theresa , Mitterbauer Gerlinde , Schloegl Ernst , Valent Peter 
Published: October 2019

Abstract
Abstract 4496
The multikinase BCR/ABL inhibitor dasatinib exerts major growth-inhibitory effects in imatinib-resistant patients with chronic myeloid leukemia (CML). Although previously reported to be a well-tolerated drug, several side effects such as pleural and pericardial effusion have been reported at a dose of 140 mg daily. Therefore, the approved standard dose of dasatinib was reduced to 100 mg daily. More recent data suggest that freshly diagnosed patients with CML treated with dasatinib at 100 mg once daily may still develop pleural effusions, but the frequency of this side effect was reported to be lower compared to patients receiving 140 mg daily dasatinib. Moreover, effusion formation was reported to be less severe, with almost no grade 3/4 events, in patients receiving 100 mg daily dasatinib. We examined a cohort of CML patients (n=13) treated with dasatinib at 100 mg or 50 mg daily, for development of severe non-hematologic side effects. Patients received dasatinib for at least 3 months (range: 3–38 months; observation period: March 2005 to June 2010). Of these 13 patients, 4 had previously received 140 mg dasatinib daily. In all 4 patients, the dose of dasatinib was reduced to 100 mg or 50 mg daily because of effusion formation. In the other 9 patients, the initial dose of dasatinib was 100 mg daily. Of these 9 patients, 2 patients in chronic phase (CP) received dasatinib as frontline therapy. Four of our 13 patients developed severe effusions (grade 3/4) while on dasatinib at 100 mg or 50 mg daily, including one CP patient receiving dasatinib as frontline therapy. In two patients, massive pleural effusions were recorded, and two developed pericardial effusion together with pleural effusions. In two patients, effusion formation required paracentesis and surgical intervention. No pre-existing cardiac or pulmonary diseases were known in these 4 patients. In 3 patients, dasatinib had to be discontinued despite treatment with low dose glucocorticosteroids and diuretics. After switching to an alternative BCR/ABL inhibitor, no further effusion formation occurred. In summary, dasatinib-treated patients are at risk for the development of pleural and pericardial effusions even when the drug is administered at a dose of 100 mg or 50 mg daily. Therefore, all patients should be examined carefully for pre-existing relevant comorbidities and cardiovascular risk factors before dasatinib is started, and all patients should have repeated chest × ray controls during long-term treatment with dasatinib.

Disclosures:
Valent: Novartis: Honoraria, Research Funding; BMS: Honoraria, Research Funding.
Imatinib Mesylate (IM) Versus IM + Low Dose of Pegylated Interferon Alfa 2a (Peg-IFN 2a) In Newly-Diagnosed Chronic-Phase (CP) Chronic Myeloid Leukemia (CML).
Authors: Meillon Luis Antonio , Campos-Cabrera Salvador , Delgado-Lopez Nancy , Fernandez-Martinez Angel A. , Medrano-Contreras Jesus , Gutierrez Guillermo , Perez-Rocha Juan F. , Terreros-Muñoz Eduardo , Gomez-Cortes Sue C. , Guerrero-Rivera Susana , Nacho-Vargas Kevin A. 
Published: October 2019

Abstract
Abstract 4497

Background:
IM 400mg daily is the current standard of treatment of CP CML. Almost 70% of patients achieve complete cytogenetic response (CCR) at 12 months (mo) (IRIS Trial) and the rate of Major Molecular Response (MMR) is 40% with IM 400mg daily (TOPS Trial). On the other hand, low doses of Peg-IFN 2a has the same efficacy and lower toxicity profile and cost as high dose, mainly when it is combined with other drugs (MRC and Hovon groups, Blood 2004; 103:4408). Combination of IM + Peg-IFN 2a or Cytarabine could increase the rate of response in CML.

Aims:
To evaluate the rates of CCR and MMR at 12 mo in Mexican patients with newly-diagnosed CP-CML treated with IM versus IM + Peg-IFN 2a.

Methods:
Between 2006 and 2008, 10 patients were randomized 1:1 and stratified by Sokal and Hasford risk groups. Cytogenetic and Molecular assesments were available for all patients. In the arm A of the study IM 400 mg daily was delivered. In the Arm B IM + Peg-IFN 2a was delivered at 400 mg daily + 90 μ g per week respectively. Peg-IFN 2a was started in the 7th week of treatment (Hochhaus A, et al ASCO 2003. Abstract #2287).

Results:
For this initial report, 10 patients were analyzed, median age was 35 years (25-55), 50% males; Sokal score was: low risk 70%, and high risk 30%. Hasford score was: low risk 40%, intermediate 30% and high risk 30%. All patients are alive with a median follow-up of 41 mo (30-48). CCR at 12 mo was obtained in 80% in both arms of treatment. MMR was obtained in 10% in patients with IM alone and 30% in patients with IM + Peg-IFN 2a. Overall the rate of primary resistance was 10% and the rate of suboptimal response was 10%. IM 400 mg was well tolerated. Peg-IFN 2a was tolerated in most patients, but needed discontinuation because increased cytopenias or other non hematologic toxicities. The median time of duration of treatment with Peg-IFN 2a was 6 mo (3-12).

Conclusions:
IM 400 mg and IM 400 mg + Peg-IFN 2a obtained similar rates of CCR. IM 400 mg + Peg-IFN 2a obtained a higher rate of MMR. The combination of IM 400 mg + Peg-IFN 2a is feasible. There is a slight increase in toxicity with this combination. Further follow-up is warranted for these patients.

Disclosures:
No relevant conflicts of interest to declare.