bluuebunny/crossref_metadata_2025 huggingface dataset:
Prognostic Factors in CBFB-MYH11 Positive AML: Trisomy 21, AGE, and t(16;16) Are Associated with Inferior Outcome.
Authors: Weisser Martin , Schnittger Susanne , Kern Wolfgang , Hiddemann Wolfgang , Haferllach Torsten , Schoch Claudia
Published: April 2020

The fusion transcript CBFB-MYH11 is the molecular correlate of inv(16)/t(16;16) and strictly associated with FAB subtype M4eo. This subgroup is associated with a favorable prognosis in AML. However, approximately 30% of the patients relapse. Our intention was to examine prognostic factors for the outcome within this subgroup. Therefore 153 CBFB-MYH11 positive AML patients were analyzed. The median age was 52 years (range 18–83), 80 patients were female, 73 were male. In 22 cases AML was therapy-related, in 131 cases a de novo AML was diagnosed. Inv(16) was detected in 138 and t(16;16) in 12 cases. In 3 cases neither inv(16) nor t(16;16) were detectable despite PCR and FISH positivity for CBFB-MYH11 suggesting cryptic rearrangements. The most frequent additional cytogenetic abnormalities were +8 (n=19), +9 (n=3), +21 (n=7), +22 (n=23). Cox regression analysis revealed that advanced age (OS: p=0.026; EFS: p=0.029) and increased CBFB-MYH11/ABL ratio at diagnosis (OS: 0.016, EFS: p=0.064) were associated with a worse prognosis. Using log rank test additional factors influencing survival were detected. These included: t(16;16) vs inv(16) (OS: n=8, censored 4, median 362 days vs n=118, censored 92, median not reached, p=0.018; EFS: n=8, censored 4, median 232 days vs n=118, censored 70, median 918 days, p=0.048) and trisomy 21 vs no additional aberrations (OS: n=6, censored 3, median 435 days vs n=74, censored 59, median not reached, p=0.024; EFS: n=6, censored 2, median 293 d vs n=74, censored 44, median 764 days, p=0.0047). Therapy related AML was associated with worse EFS than de novo AML (n=16, censored 6, median 371 days vs n=112, censored 70, median 1179 days, p=0.0167) and there was a trend towards worse OS (p=0.157 n=16, censored 10, median 764 days vs n=112, censored 88, median not reached). A multivariate analysis including t(16;16), age, CBFB-MYH11/ABL ratio, therapy related AML and +21 as covariates revealed t(16;16) and age as independent factor for OS (p=0.014 and p=0.015, respectively) and age, t(16;16), and +21 as independent factors for EFS (p=0.047, p=0.013, and p=0.016, respectively). There was no evidence that the additional aberrations +22 or +8 had an influence on survival. Taken together our data suggest that t(16;16) as compared to inv(16), trisomy 21 and age are associated with worse prognosis in patients with CBFB-MYH11 positive AML.
Successful Induction of Hematologic and Cytogenetic Remission with Gemtuzumab Ozogamicin (Mylotarg(R)) in Patients with Myeloid Blast Crisis of Chronic Myelogenous Leukemia in Refractory to Imatinib Mesylate and Chemotherapy.
Authors: Weng Jingyu , Du Xin , Wu Shuijing , Lu Zesheng , Huang Zilu , Lin Wei
Published: April 2020

Introduction The treatment options for patients with chronic myeloid leukaemia (CML) in blast crisis phase are limited, especially to those were refractory to imatinib mesylate. We report the achievement of hematologic and molecular completely remission after the use of gemtuzumab ozogamicin( mylotarg) in two patients with CD33-positive CML-BC who were refractory to chemotherapy and imatinib mesylate(Gleevec). Two patients(male,62yr; female, 40yr), Before received Mylotarg, they were administrated imatinib mesylate(600mg per day) more than three months and receiveed combination chemotherapy, but they were refractory and their disease progressed. Then, they received Mylotarg as a 2-hour intravenous infusion, at a dose of 9 mg/m2, at 2-week intervals for three doses. One of the patient was treated with Mylotarg combinated MA(Mitoxantrone 5mg, d1–4; cytarabin 100mg/M2,d1–7). After the first cycle Mylotarg, two patients achieved a complete hematologic and molecular remission, bcr/abl fusion gene negative detected by FISH, real time PCR bcr/abl ratio 0.03. They have been alive and continuous complete remission.

CONCLUSION: Mylotarg alone or combinated with chemotherapy to patients with CD33-positive CML-BC in refractory to imatinib mesylate can obtain complete hematologic and molecular cytogenic remissions with what appears to be a favorable safety profile.
Serum Protein Profiling Distinguishes BCR-ABL+ CML from Normal and Neutrophilia Cases.
Authors: Mohamedali Azim M. , Sahu Satyaji , Thomas Nicholas Shaun B. , Mufti Ghulam J.
Published: April 2020

We sought to identify additional biomarkers for chronic myelogenous leukaemia (CML) that could be an aid to early diagnosis and also yield novel antigens for immunotherapy. To this end, we screened patient serum samples at presentation against hematologically normal controls as well as patients with neutrophilia using Surface Enhanced Laser Desorption/Ionization technology (SELDI; Ciphergen ProteinChip series 4000). A total of 84 retrospective and prospective serum samples were analysed: presentation −28, reactive neutrophilia (>15x109 neutrophils/L) − 24 and hematopoietic normal controls − 33. Patients were initially screened by routine cytogenetics and in some cases with qPCR for the BCR-ABL breakpoint. The sera samples were evaluated on 4 different array surfaces and the Immobilised Metal Affinity (IMAC) array was chosen as it bound serum proteins that distinguished CML from normal controls. As little as 1 μl serum was sufficient for each analysis. Biomarker artefacts due to variations in sample collection procedures were ruled out by analysing sera (n=4) from each group at the time of collection and 3 and 6 hours post collection. There were no significant differences in any of the biomarkers at any of the time points. The spectrum of proteins obtained from each of the 84 serum samples was averaged from duplicate runs per experiment. Using the Ciphergen Express program, a panel of 5 proteins were significantly differentially expressed in CML versus the reactive neutrophilia and normal hematopoietic controls (p<0.001). These proteins were identified by a combination of purification techniques using Q HyperD F columns, desalting using reverse phase C-18 beads and isolating the biomarker by 1D-SDS PAGE. The biomarkers were identified by peptide mass fingerprinting and confirmed by Tandem MS sequencing. These were

Albumin fragment − 2.8Kd (p< 3.5 x 10−5, ROC=0.78), Fibrinogen fragments − 5.3Kd (p< 6.25 x 10−10, ROC=0.07) and 5.9Kd (p< 9.6 x 10−8, ROC=0.14), Complement 3a precursor fragment − 8.9Kd (p< 0.0015, ROC= 0.70), Platelet basic protein precursor − 10.2Kd (p< 1.5 x 10−4, ROC=0.73) and Lysozyme − 14.6Kd (p=0, ROC=0.92).

Biomarkers 3, 4 and 5 were also verified by antibody capture experiments using NP-20 arrays. In a blinded test set of sera, CML, normal and neutrophilia samples were correctly classified 27/28 (96%), 32/32 (100%), 20/24 (83%) respectively using a combination of the 5.3Kd, 10.2 Kd and the 14.6 Kd markers (Biomaker Pattern software). The algorithm correctly classified 21 new samples as CML (7/8) and control (10/13). The 1/8 CML was misclassified for technical reasons. Therefore, a small number of serum biomarkers in as little as 1 μl serum can be used to distinguish between patients with CML and neutrophilia or hematopoietic normal controls. Similar analyses may be applicable to other more heterogeneous hematological malignancies.
The Induction of Dendritic Cells with IFN-alpha and GM-CSF from Bone Marrow Mononuclear Cells from Patients with Chronic Myeloid Leukemia.
Authors: Zhen Shuier , Jin Jie , Tong Xiangmin
Published: April 2020

Chronic myelogenous leukemia (CML) is a malignant myeloproliferative disease arising from the clonal expansion of a stem cell with the typical Philadelphia (Ph) chromosome cytogenetic abnormality. IFN-a has been proven to be effective for patients in the chronic phase of myelogenous leukemia (CML), yet the mechanisms of the antitumor action of these cytokines are still a matter of debate. Dendritc cells (DCs) are potent antigen-presenting cells that prime effective T-cell response aginst tumour antigens. Recent studies have shown that IFN-a can exert a variety of effects on dendritic cells (DCs), which may play an important role in the induction of an antitumor immunity. Human DCs can be generated in vitro from peripheral blood(PB) monocytes or from CD34+ haematopoietic precursor cells in culture medium containing human granulocyte macrophage-colony stimulating factor (GM-CSF), IL-4 and some other cytokines. Previous studies have shown a new effective protocol for the generation of human DCs from unseparated BM aspirate cells with excellent functional capacity of antigen uptake and of stimulating naive and memory T cell responses superior to that of DCs from peripheral blood(PB) monocytes. We, therefore, explored whether treatment with IFN-a may influence the CML bone marrow mononuclear cells(BMMNCs) derived DCs in vitro. Treatment BMMNCs of 12 patients with CML in chronic phase with IFN-a+rhGM-CSF(IFN-a-DC) generated DCs with more mature phenotype properties expressing higher of CD80,CD86,HLA-DR,CD83 compared to the CML- BMMNCs treated with rhGM-CSF+IL-4(IL-4-DC). And in parallel with phenotypes, IFN-a-DC also showed more effective than IL-4-DC in eliciting an allogeneic mixed lymphocyte reaction by MTT assay. FISH confirmed the DCs of both groups were leukemic origin. These findings demonstrate that IFN-a promotes the differentiation/maturation of DCs derived from BMMNCs of patients with CML in vitro, these studies also broaden the clinical scope of IFN-a as a promising agent in the immunotherapy of CML.
Effects of STI571 on the Development of Dendritic Cells Derived from Bone Marrow Mononuclear Cells from Patients with Chronic Myeloid Leukemia.
Authors: Jin Jie , Zheng Shuier , Tong Xiangmin
Published: April 2020

Mature dendritc cells(DCs) are potent antigen-presenting cells that prime effective T-cell response aginst tumour antigens. However, in the tumour microenviroment the differentiation and maturation of DCs is always suppressed by many factors, such as VEGF, IL-10,TGF-βet al. Though DC derived from CML patients have been shown to process and present endogenous BCR-ABL to CD4+ T cells, the function of DC may be abnormal in CML, which have been shown differences in cytoskeletal organisation and impaired migration, and a reduced capacity to capture and present nonleukaemic antigen, and less effective than normal DC in eliciting an allogeneic mixed lymphocyte reaction. STI571(trade name Glivec or Gleevec) is a specific inhibitor of Abl tyrosine kinase and has been a first- line agent for the treatment of chronic myelogenous leukemia (CML). Previous studies have found that defective blood dendritic cells in chronic myeloid leukemia correlate with high plasmatic VEGF and can be partially normalized by STI571. We, therefore, explored whether treatment with STI571 may influence the CML -derived DC in vitro. Treatment bone marrow mononuclear cells(BMMNCs) of 11 patients with CML in chronic phase with STI571+rhGM-CSF+IL-4(STI571-DC) generated DCs with more mature phenotype properties expressing higher of CD80,CD86,HLA-DR,CD83 compared to the CML- BMMNCs treated with rhGM-CSF+IL-4(IL-4-DC). And in parallel with phenotypes, STI571-DC also showed more effective than IL-DC in eliciting an allogeneic mixed lymphocyte reaction by MTT assay. FISH confirmed the DCs of both groups were leukemic origin. To explore whether the positive effects of STI571 on CML-DC may attributed to negative factors of DC differentiation/maturation suppressed by STI571, media were tested for VEGF concentrations by using the sandwich enzyme-linked immunosorbent assay (ELISA). As a result, the concentration of VEGF was dramatically reduced in STI571-DC. These findings demonstrate that treatment with STI571 promotes maturation and enhances the antigen-presenting capabilities of CML- BMMNCs derived DCs, these studies also offer a more comprehensive understanding of the role of STI571 in the immunotherapy of CML.
Sphingosine Kinase 1 Is Involved in Proliferation and Survival of Chronic Myeloid Leukemia Cells.
Authors: Ricci Clara , Onida Francesco , Soligo Davide , Lambertenghi-Deliliers Giorgio , Ghidoni Riccardo
Published: April 2020

Sphingosine Kinase 1 (SK1) is a key enzyme of the sphingolipid metabolism. By phosphorylating sphingosine (Sph), SK1 induces accumulation of sphingosine 1 phosphate (S1P). As a second messenger, S1P stimulates proliferation and survival and protects cells against ceramide-induced apoptosis. Although oncogenic activity of SK1 has been demonstrated in a variety of solid tumors, its role in leukemic cells growth remains unclear. Recently, SK1 overexpression has been reported to represent an oncogenic event during erythroleukemia progression (Lescolan E. et al, Blood 2005). We focused on Chronic Myeloid Leukemia (CML), a hematopoietic disorder characterized by the presence of the Philadelphia (Ph) chromosome that leads to the expression of the chimeric protein Bcr/Abl. Through its constitutive tyrosine kinase activity, Bcr/Abl is recognized as the pathogenetic event that causes CML and thus the basis for the development of specific inhibitors. Among these, Imatinib Mesylate (IM) represents the more successful drug for CML treatment. However, most patients who are treated in advanced stages of the disease relapse after an initial response to IM. Therefore, the existence of a functional link between Bcr/Abl and SK1 would provide new strategies to overcome acquired resistance to IM in CML. To test the hypothesis that SK1 contributes to the growth and survival of leukemic cells, we first verified that the kinase transcript is expressed in a panel of CML cell lines [AR230 (p230Bcr/Abl+), K562 and RWLeu4 (both p210Bcr/Abl+)], although at different levels. Then the effect of pharmacological inhibition of SK1 was assessed by using a competitive inhibitor (SKI, SK Inhibitor, Calbiochem) of the ATP-binding site of SK1. Cells were treated with increasing doses (1 to 50 μM) of SKI for up to 72 hrs and the WST-1 assay was performed. A dose- and time-dependent anti-proliferative and cytotoxic effect was observed (IC50: 7.02 μM for AR230, 7.06 μM for K562 and 35.8 μM for RWLeu4 cells). Surprisingly, up-regulation of SK1 transcript was detected. Finally, when the Eosinophilic Leukemia cell line Eol-1 (FIP1L1-PDGFRα+) was exposed to SKI under the same experimental conditions of CML cells, an even more potent anti-proliferative and cytotoxic effect was observed, indicating a higher sensitivity of these cells to SKI (IC50: 0.43 μM). Altogether our results suggest involvement of SK1 in the control of growth and survival of myeloid leukemia cells of different origin. To address the question whether a functional link between SK1 and Bcr/Abl occurs, we are now investigating if Bcr/Abl expression and phosphorylation status are modulated following treatment with SKI and conversely if SK1 expression and activity are affected by treatment with IM.
Chronic Myeloid Leukemia during Pregnancy: Report of Management and Outcome in Seven Patients.
Authors: Matias Kleber , Matias Carolina , Freire Ana Dulce , Azevedo Alita Andrade
Published: April 2020

The occurrence of leukemia during pregnancy is very rare with an estimated incidence of one per 100,000 pregnancies annually. It has been estimated that during pregnancy most leukemias are acute: two thirds are myeloid (AML) and one third are lymphoid (ALL). Chronic myeloid leukemia (CML) is found in less than 10% of leukemias during pregnancy and chronic lymphocytic leukemia (CLL) is extremely rare. The management of CML during pregnancy is a difficult problem because of the potential effects of the therapy on the mother and fetus. Since the disease has an initial chronic phase, it is usually managed conservatively during pregnancy, while an aggressive approach, such as bone marrow transplantation, may be considered after delivery. A limited number of cases described successful treatment modalities of CML during pregnancy including leukapheresis, hydroxyurea (HU) and interferon (IFN). We report nine cases of pregnancy in seven chronic myeloid leukemia patients, giving birth healthy children in a single institution from 1979 to 2005. In four cases the diagnosis of CML was made on prebirth period in routine blood testing, and five pregnancies developed during the course of disease. Four of the pregnancies were found in the first trimester, four in the second and one in the third. Median age of patients was 21 years (range 18–30years). All patients were Ph1 positive and the leucocyte count ranged between 45 to 336 x 109 /L. Table 1 shows treatment performed in patients before and during pregnancy. Patients 4 and 7 had a subsequent pregnancy despite the use of contraceptive methods, both diagnosed in the first trimester. Hydroxyurea was stopped during pregnancy. Delivery was performed by caesarean section in 5 cases and by spontaneous vaginal delivery in 4 cases. All infants’ examination and blood counts were normal and there were no perinatal or maternal complications. In june 2005, two new cases of pregnant CML patients were seen at our institution. One of them was being treated with imatinib, and the other without treatment at the moment of pregnancy. They will be managed only with leukapheresis. Our data suggest that exposure to IFN and HU during pregnancy is probably not associated with a significantly increased risk for malformations, however leukapheresis can be considered for treatment of CML during pregnancy because of the lack of teratogenic and other adverse effects in patients who tolerate and respond to the procedure.

Cases Age (years) CML diagnosis Pregnancy diagnosis Trimester of pregnancy Treatment before pregnancy Treatment during pregnancy 1 18 Sep/1993 Oct/1993 First None HU 2 30 Dec/1996 Jan/1997 Third None Leukapheresis 3 18 Dec/1993 Nov/1995 First IFN stopped IFN 4 21 Sep/1995 Sep/1995 Second None HU 5 21 Jul/1994 Aug/1994 Second HU stopped HU 6 27 Jan/1979 Jan/1979 Second None Busulphan - 2 months 7 18 Dec/1995 Oct/1996 Second HU HU
Study on the Burden of Abnormal Hematopoietic Clone of the Patients with Myelodysplastic Syndromes.
Authors: Shao Zonghong , Wang Huaquan , Shi Jun , Cao Yanran , Liu Hong , Bai Jie , Tu Meifeng , Xing Liming , Liu Shihe , Sun Juan , Jia Hairong
Published: April 2020

Background Myelodysplastic syndromes (MDS) is a group of clone haematopoietic stem cell diseases. The burden of abnormal hematopoietic clone plays key roles in the development of this disease and needs to be further studied quantitively. Methods The ratio of the counted bone marrow cells with abnormal chromosomes to the total counted bone marrow cells was regarded as the index of MDS clone burden. The disease severity related parameters including white cell count, hemoglobin, platelet count, lactate dehydrogenase level, bone marrow blast, myeloid differentiation index, the ratio of cFU-GM to CFU-GM, micromegakaryocyte, transfusion, IL-2, TNF, CD4+ and CD8+ T cells of MDS patients were assayed and the correlations between those parameters and MDS clone burden were also analysed. Results The clone burden of MDS patients was (67.4±36.2)%. MDS clone burden correlated positively with bone marrow blasts(r =0.483, P=0.012), negatively with hemoglobin level(r = −0.445, P= 0.023); The number of blasts, hemoglobin and erythocytes in high clone burden(>50%) and low clone burden(≤50%) groups were (7.78±5.51)% and (3.45±3.34)%(P=0.035), (56.06±14.28)g/L and (76.40±24.44)g/L(P=0.013), (1.82±0.12)×1012/L and (2.32±0.21)×1012/L(P=0.034)respectively. CD4+ T lymphcytes of MDS patients and normal controls were (274.18±71.85)×106/L and (454.82±205.88)×106/L(P=0.012)respectively. CD8+ T lymphcytes of MDS patients and normal controls were (240.45±150.01)×106/L an (305.27±145.14)×106/L(P=0.317)respectively. The serum level of interleukin 2 of MDS patients and normal control group were (6.29±3.58)ng/ml and (3.11±1.40)ng/ml (P=0.002) respectively. The serum level of tumor necrosis factor of MDS patients and normal control group were (2.42±1.79)ng/ml and (1.68±0.69)ng/ml(P=0.124)respectively. The ratio of CD4 to CD8 in high clone burden MDS patients was (1.90±0.52), and that in low clone burden patients was (0.97±0.44)(P=0.022). Conclusion The quantitive clonal karyotype abnormalities and deficient T cell immunity are important parameters for evaluating MDS severeity and prediciting it’s progression.
Exisulind Selectively Induces Apoptosis Via C-Jun Kinase Activation in MDS and sAML/MDS.
Authors: Czibere Akos , Prall Wolf C. , Knipp Sabine , Kuendgen Andrea , Zerbini Luiz F. , Germing Ulrich , Haas Rainer , Libermann Towia A. , Gattermann Norbert , Aivado Manuel
Published: April 2020

Treatment of patients suffering from myelodysplastic syndromes (MDS) is difficult and frustrating, particularly when in progression or already progressed to secondary acute myeloid leukemia (sAML). In these cases, the bone marrow contains an increasing number of immature cells, which are characterized by reduced apoptotic and increased proliferative activity. Therefore, pro-apoptosis has recently been proposed as a novel therapeutic approach. Exisulind is a potentially pro-apoptotic agent, and therefore, we investigated its influence on proliferation, differentiation, cell cycle and apoptosis in peripheral blood-derived CD34+ stem cells (PBSC) obtained from 10 patients suffering from refractory anaemia with excess of blasts in transformation (RAEB-T), two sAML/MDS cell lines, one de-novo AML cell line and healthy CD34+ bone marrow cells. Incubation of sAML/MDS cells with Exisulind clearly inhibited colony formation in the CFU-assays. Interestingly, Exisulind did not alter the percentages of sAML/MDS cells in G1-, S-, G2-, M-phase, but reduced proliferation and induced apoptosis in this cell type. Likewise, in 8 out of 10 RAEB-T samples Exisulind significantly inhibited proliferation and increased the rate of apoptotic cells. Exisulind had no effect on de-novo AML or normal CD34+ cells. We detected increased c-Jun NH2-terminal kinase activity in sAML/MDS cells treated with Exisulind. Adding a specific JNK-inhibitor to Exisulind-treated sAML/MDS and RAEB-T cells partly abrogated apoptosis, thus proving that Exisulind-mediated apoptosis in sAML/MDS and RAEB-T cells is dependent on JNK activation. We conclude that JNK is one mediator of apoptosis in MDS cells treated with Exisulind. Moreover, our data strongly suggests to explore the potential use of Exisulind as a novel, pro-apoptotic therapy for patients with RAEB-T and sAML/MDS.
Study of Intra-Venous Homoharringtonine (HHT) in the Treatment of Myelodysplastic Syndrome (MDS).
Authors: Borthakur Gautam , Kantarjian Hagop , Estey Elihu , Verstovsek Srdan , Garcia-Manero Guillermo , Cortes Jorge
Published: April 2020

Homoharringtonine (HHT) is a plant alkaloid that inhibits protein synthesis, induces differentiation and apoptosis of myeloid cells. We initiated a phase II study of intra-venous HHT in patients (pts) diagnosed with MDS by WHO criteria to assess safety and efficacy of HHT in this setting. Responses were recorded according to the MDS response criteria defined by NCI working group. Pts received HHT 2.5 mg/m2 by continuous 24-hour intravenous infusion daily for 7 days every 4 weeks until a) a complete remission (CR) was achieved, or b) failure to achieve a CR after 3 courses of therapy or c) unacceptable toxicities developed. The dose was increased to 3.0 mg/m2 for pts with no response after the 1st course. Nine pts have been enrolled (6 male and 3 female). Median age was 70 yrs (range 55 to 84 yrs) and 4 pts had secondary MDS. Four pts had refractory anemia with excess blasts-1 (RAEB-1), 3 had RAEB-2, 1 had chronic myelomonocytic leukemia (CMML-1) and 1 had MDS/myeloproliferative disorder (MDS/MPD). IPSS score was intermediate-1 in 1 pt, intermediate-2 in 7 pts and high in 1 pt. Seven pts (78%) received only one cycle of HHT. One (11%) pt had a CR (including cytogenetic CR) that lasted for 67 days, 5 pts discontinued treatment due to lack of response and1 pt with stable disease decided to withdraw from the study. After a median follow-up of 54 days (range, 37 to 145 days) 2 pts have died, one on day 37 due to intracranial bleed secondary to thrombocytopenia and one on day 39 due to invasive fungal infection. Grade 2 toxicities included fatigue (4 pts), diarrhea (3 pts) and nausea (3 pts). Grade 3 cytopenias were seen in all pts and grade 3 nausea in 1 pt. Three pts had neutropenic fever. Treatment during first course was interrupted after 5 days of infusion in 1 pt due to grade 3 congestive heart failure (CHF). This pt received his second course of treatment without recurrence of CHF. In conclusion, HHT has modest activity in intermediate to high risk MDS. After first interim analysis, the study meets criteria for continued accrual.
Reversal of Myelofibrosis in a Patient with Low Risk Myelodysplastic Syndrome on Revlimid Therapy.
Authors: Besa Emmanuel C. , Swami Vanlila , Roda Paul , Laudadio Mary Ann , Knight Robert , Zeldis Jerome
Published: April 2020

The observation of increasing myelofibrosis has been interpreted as a sign of progression in the course of low risk patients with myelodysplastic syndrome (MDS). We report here a 64 year old, Caucasian woman who was diagnosed to have refractory anemia in 5/19/03 with a bone marrow findings showing hypercellularity of 95%, erythroid dysplastic changes with 1% myeloblasts with no ringed sideroblasts. Her cytogenetic study show normal chromosomes and she had an IPSS score of 0. However, she began to require red cell transfusions with no response to EPO and G-CSF therapy for 5 months when she was entered into an open label, multi-center clinical trial using Revlimid at 10 mg/day given orally in tablet form. Her growth factors were discontinued 4 weeks before entry and a baseline bone marrow study on 10/7/04 show a 90% cellular marrow with same abnormalities. In addition, she now has developed a grade III/IV diffuse reticulin fibers as shown by silver stains indicating increased myelofibrosis. She tolerated the treatment well with very little side effects and by second month of therapy, she had her last red cell transfusion and her hemoglobin level started to increase from 8.3 to 9.7 g/dl. Other than a mild neutropenia with ANC between 800 to 900/μL, she tolerated treatment well. She attained a normal hemoglobin of 12 g/dl at the 4th month. Repeat BM done at this time shows a normal cellularity of 50% but erythroid cells show persistence of dysplasia but overall fibrosis has reversed to Grade I/IV. She currently remains in remission 21 months on Revlimid treatment. Demonstration of reversal of myelofibrosis in MDS indicate that this drug may be effective in primary myelofibrosis and agnogenic myeloid metaplasia.
Phase I/II Trial of Bortezomib + CHOP-Rituximab in Diffuse Large B Cell (DLBCL) and Mantle Cell Lymphoma (MCL): Phase I Results.
Authors: Leonard John P. , Furman Richard R. , Cheung Ying-Kuen K. , Feldman Eric J. , Cho Hearn J. , Vose Julie M. , Nichols Gwen , Glynn Patricia W. , Joyce Maureen A. , Ketas Jamie , Ruan Jia , Carew John , Niesvizky Ruben , LaCasce Ann , Chadburn Amy , Cesarman Ethel , Coleman Morton
Published: April 2020

BACKGROUND: Bortezomib (PS-341, VelcadeR) is a proteasome inhibitor with anti-tumor activity in B cell malignancies including MCL. These effects, which in part may be mediated via NF-kappaB pathways, could sensitize tumor cells to standard chemotherapy-based treatment regimens and enhance efficacy. We report phase I findings from a phase I/II trial of dose-escalated bortezomib + standard CHOP-rituximab in DLBCL and MCL patients.

METHODS: Thirty-five patients have been enrolled and initiated treatment, with the first 20 subjects comprising the phase I group. Patients received CHOP-21 + rituximab (375 mg/m2 each cycle) plus bortezomib at 0.7 mg/m2 (Arm 0, n=4), 1.0 mg/m2 (Arm 1, n= 9) or 1.3 mg/m2 (Arm 2, n=7 in phase I, then all subsequent patients in phase II) on days 1,4 of each cycle. Phase 1 dose assignments were conducted according to the continual reassessment method as patients entered the study.

RESULTS: Phase I included 16 DLBCL and 4 MCL patients. Median age was 65 years (range 29–84) and all patients were treatment-naïve except for 1 MCL subject who had received prior local radiotherapy. Fourteen subjects (70%) had stage IV disease at study entry, and 11 (55%) had elevated serum lactate dehydrogenase (LDH). Patients generally had unfavorable baseline international prognostic index (IPI) scores of 2 in 7 subjects (35%) and 3 or 4 in 10 subjects (50%). Median followup of the phase I cohort is 9 months (range 5 – 18 months). Treatment was generally well tolerated. Peripheral neuropathy occurred in 15 subjects (75%), with 65% grade 1, 5% grade 2 and 5% grade 3. Grade 4 hematologic toxicity (platelets and/or white blood cells) was observed in 7 subjects (35%). Three subjects did not complete therapy (1 withdrawal by patient request with subsequent death, 1 due to dose-limiting pulmonary toxicity and 1 with concurrent lyme meningitis). Intent to treat overall response rate (n=20) is 95% with 80% CR/CRu and 80% of subjects currently alive without progression.

CONCLUSIONS: Bortezomib can be administered with acceptable toxicity in conjunction with CHOP-R chemotherapy. Further safety and efficacy analyses, as well as molecular correlative studies, are ongoing with both the phase I and II cohorts. Preliminary findings with the combination regimen in this adverse prognosis group of patients are encouraging.
Down Regulation of hTERT Is an Important Mechanism in Apoptosis of MUTZ-l Cells Induced by As2O3.
Authors: Jin Jie , Xu Weilai , Qian Wenbin
Published: April 2020

Myelodysplastic syndromes (MDS) are a heterogeneous group of clonal hematopoietic disorders characterized by ineffective hematopoiesis resulting in peripheral cytopenia and by increased progression to acute myeloid leukemia (AML). In this study, MDS cell line MUTZ-1 cells were treated with As2O3 to demonstrate the effects of As2O3 on MDS. We found As2O3 inhibited the proliferation of MUTZ-1 cells in a dose- and time-dependent manner. As2O3 induced apoptosis in MUTZ-1 cells via activation of caspase8, 3 and subsequent cleavage of the DNA repair enzyme pol(ADP-ribose) polymerase(PARP). Subsequently, reduction of NF-kB activity was another important mechanism of As2O3 induced apoptosis in MUTZ-1 cells. Our results also showed that the expression of hTERT mRNA of MUTZ-1 cells was down-regulated by As2O3. Except NF-κB, activity of other nuclear transcription regulator of hTERT, SP-1 and AP-1 were inhibited meanwhile using gel shift assay. Moreover, we further found that NF-κB inhibitor, PDTC, couldn’t activate capase8 but inhibited growth of MUTZ-1 cells and down-regulated hTERT mRNA expression. In addition, caspase8 inhibitor can’t block the reduction of NF-κB activity by As2O3. We concluded that As2O3 could induce apoptosis in MUTZ-1 cells by two independent ways, one is caspase8,3 and PARP activation, the other is inhibition of NF-κB activity subsequent down-regulation of hTERT. hTERT and NF-κB were two important molecular targets in apoptosis induced by As2O3. As2O3 down-regulated hTERT expression via inhibition of NFκB, SP-1 and AP-1.
The Establishment of Mice Model with Myelodysplastic Syndrome Cell Line MUTZ-1 Cell.
Authors: Lu Jie , Jin Jie , Xu Weilai , Chen Zhimei , Ding Wei
Published: April 2020

Objective

To establish myelodysplastic syndrome cell line MUTZ-1 cell mice model.

Methods

75 SCID mice and 10 BALB/CA-node mice were studied in this experiment. MDS-REBT cell line MUTZ-1 cells were cultured in vitro and 1 x108 /ml cell were subcutancously implanted in 4~6-week-old First-Generation SCID mice and BALB/CA-node mice respectively. The subcutancous tumor cells from First-Generation MDS-REBT cell line MUTZ1 cells mice model were respectively implanted in Second-Generation SCID mice and BALB/CA-node mice. The latent period and the rate of subcutancous tumor formation was observed, and tumor size was measured by the MTD twice a week. The biological characteristics of the subcutancous tumor cells were checked and evaluated by the methods of cell morphology and pathology and histopathology and immunology by flow cytometer, chromosome analysis and immunohistochemistry stain. It has been experimented tentatively that arsenic trioxide (AS2O3) (with 7.5mg/g mouse/d x 3d) was injected into abdominal cavity in 10 SCID mice to observe the drug effect on tumor formation in vivo.

Results

The rate of the subcutancous tumor formation was 98.6% (70/71)in SCID mice and 62.5%(5/8)in BALB/CA-nude micerespectively,(P=0.0027). The latent period of the subcutancous tumor formation were 10~17d (median 12d) in 61 SCID mice without injected arsenic trioxide and were 26~31d (median 29d) in 10 SCID mice with arsenic trioxide (with 7.5mg/g mouse/d x 3d) injected into abdominal cavity respectively. The latent period of the subcutancous tumor formation in the group of SCID mice injected by AS2O3 was significantly longer than that of non-injected by AS2O3(Z=5.339, P<0.001), which showed that AS2O3 has probably a inhibition effect on the subcutancous tumor growth of SCID mice model in vivo. The results showed the subcutancous tumor cells of mice model have the biological characteristics of a Hum-MDS-REBT cell line MUTZ1 cell and were anthropo- source. The metastases in liver were found 8 weeks after implanted. HE staining on paraffin sections showed that the tumor cells were microscopic observed on liver and spleen and kidney and medullary cavity of bone and skin. Immunohistochemistry staining showed the subcutancous tumor cells of the mice model have the overexpression of P53 protein, which cued that these tumor cells has the P53 mutation and showed human tumor propert.

Conclusion

A human myelodysplastic syndromes-REBT cell line MUTZ1 cell-severe combined immunodeficiency (SCID) mice model was successfully established, which could be used for the advanced investigation of drug experiment as an animal model system in vivo.
Cytogenetics in Pediatric MDS - Data of the EWOG-MDS 98 Study.
Authors: Michalova K. , Beverloo H. B. , Michaelsen G. , Schlegelberger B. , Harbott J. , Haas O. A. , Kerndrup G. B. , Sainati L. , Bergstraesser E. , Hasle H. , Starý J. , Trebo M. , Van den Heuvel-Eibrink M. M. , Zecca M. , Noellke P. , Fischer A. , Niemeyer C. M. , Betts D. R.
Published: April 2020

Myelodysplastic syndrome (MDS) represents a heterogeneous group of hematopoietic stem cell disorders. Although the precise pathogenesis of MDS remains to be clarified, cytogenetic abnormalities appear to be involved in the disease mechanism and are considered an important factor in predicting clinical outcome. Here, we report the outcome of conventional cytogenetic analysis performed at diagnosis in 300 children with MDS who were enrolled in the prospective study EWOG-MDS 98. The patients were divided into 3 clinical groups: primary refractory cytopenia (RC) (n=142), primary advanced MDS (aMDS) (n=94), and MDS secondary to chemo- or radiation therapy, bone marrow failure disorders or familial disease (sMDS) (n=64). A full cytogenetic result was obtained in 263 cases (87.7%) including 116 (44.1%) with an abnormal clone. Comparing the 3 diagnostic subgroups, there was a statistically significant difference in the frequency of abnormalities observed. In RC, only 25 of 116 patients with successful analysis had a detectable clone that showed monosomy 7 as sole aberration in the majority of cases (52%). In addition, this subgroup also showed the greatest rate of failure to obtain a karyotypic result. In contrast, 42 (65.6%) of the sMDS cases were cytogenetically abnormal with a broad spectrum of aberrations including 16 patients (25%) with a complex karyotype (≥4 abr.). The presence of karyotypic evolution was also more frequently observed in this group. The overall karyotypic results showed monosomy 7 to be present in 50% of all abnormal cases, a figure that greatly exceeded the prevalence of the other frequent non-random aberrations of −5/del(5q), +8, del(7q), and +21, each of which occurred in 5–10% of abnormal cases. Despite the small numbers of these latter cases, clear trends of karyotypic events occurring in a particular clinical subgroup were demonstrated in the form of +8 with aMDS and −5/del(5q) with sMDS. The clinical and cytogenetic subgroups were further analyzed for correlations with gender and age. Whereas no clinical group showed any correlation between gender and karyotype, in RC and aMDS it was shown that patients with monosomy 7 were significantly younger than those with another karyotypic result (median 3.2 vs. 10.4 yrs and 5.2 vs. 11.0 yrs, respectively). The results demonstrate that while a general karyotypic picture can be defined in childhood MDS, future studies need to clearly differentiate between individual clinical subgroups.
No Evidence for Constitutively Activated FLT3 in Juvenile Myelo-Monocytic Leukemia.
Authors: de Vries Andrica C.H. , Stam Ronald W. , Schneider Pauline , Niemeyer Charlotte M. , van Wering Elisabeth R. , Kratz Christian , Haas Oskar A. , den Boer Monique L. , Pieters Rob , van den Heuvel-Eibrink Marry M.
Published: April 2020

Activating FLT3 mutations have been identified as prognostic factors in multiple myeloid malignancies. Recent studies have demonstrated that ligand-independent activation of FLT3 can also result from overexpression of wild-type FLT3. In addition, ligand-dependent activation has been observed in leukemic cells co-expressing FLT3 ligand (FLT3L), resulting in autocrine FLT3 signaling which is independent of FLT3 mutations. In Juvenile Myelo-Monocytic Leukemia (JMML), FLT3 internal tandem duplications (FLT3/ITDs) mutations affecting the tyrosine kinase domain (TKD) are rare. However, no data are yet available on the frequency of expression levels of FLT3 and FLT3L in JMML. If activated FLT3 occurs in JMML, one could imagine that these patients might benefit from treatment with small molecule FLT3 inhibitors, especially as to date the curative treatment of JMML is limited to allogeneous bone marrow transplantation.

In 36 JMML patients FLT3 and FLT3L mRNA levels were assessed using real-time quantitative PCR (Taqman). Furthermore these samples were screened for the presence of activating FLT3/ITDs and FLT3/TKD mutations. MTT assays were performed to assess the response of JMML cells to the known FLT3 inhibitor PKC412 (Novartis). FLT3 appeared to be expressed only at basal levels and FLT3L expression was very low. In none of the 36 JMML samples FLT3/ITDs or TDK mutations were found, consistent with the observation that PKC412 was not cytotoxic in JMML samples (n=12), in contrast to leukemic cells of children with ALL which carried an activated FLT3. These data suggest that constitutively activated FLT3 does not occur in JMML. Therefore targeting FLT3 by tyrosine kinase inhibitors like PKC412 is unlikely to be effective in JMML.
Revised International Prognostic Index (R-IPI) Is a Better Predictor of Outcome Than the Standard IPI for Patients with Diffuse Large B-Cell Lymphoma (DLBCL) Treated with Rituximab and CHOP (R-CHOP).
Authors: Sehn Laurie H. , Chhanabhai Mukesh , Fitzgerald Catherine , Gill Karamjit , Hoskins Paul , Klasa Richard , Savage Kerry , Shenkier Tamara , Sutherland Judy , Wilson Kenneth , Gascoyne Randy D. , Connors Joseph M.
Published: April 2020

Background: The IPI, which was developed prior to the availability of rituximab, remains the primary clinical tool used to predict outcome for patients with DLBCL. Since clinical trials investigating the use of rituximab and chemotherapy in this patient population have been confined to either elderly or young patients exclusively, the utility of the IPI in the era of immuno-chemotherapy remains unknown.

Methods: We performed a retrospective analysis to assess the predictive value of the IPI in an unselected population of patients with DLBCL treated with R-CHOP. Patients were identified using the Lymphoid Cancer Research Database of the British Columbia (BC) Cancer Agency. We included all patients ≥16 years of age who were newly diagnosed with DLBCL prior to Jan 15, 2005, and were treated in BC with an R-CHOP protocol. Patients were excluded if they were HIV positive, had evidence of an active second malignancy or an underlying indolent lymphoma.

Results: 365 patients were identified. Patient characteristics were as follows: median age 61 y (16–90); male, 61%; advanced stage III/IV, 59%; elevated LDH, 54%; PS≥2, 40%; >1 extranodal site, 35%. Central pathology review was performed on 95% of patients; 324 DLBCL, 36 PMBCL, 5 other. All patients received R-CHOP; 9% were treated with R-CHOP x 3 and involved field radiation therapy for limited stage disease, remaining patients received 6–8 cycles of R-CHOP. Median follow-up for living patients is 22 months. Overall, the IPI remains predictive for both progression-free survival (p<0.0001) and overall survival (p<0.0001). However, the IPI no longer permits separation between the two low risk subgroups (low v low-intermediate) or between the two high risk subgroups (high-intermediate v high). (see Table) An alternate grouping of the IPI is proposed (R-IPI) that identifies three distinct prognostic groups. (see Table and Figure)

Conclusions: The IPI remains predictive in the era of immuno-chemotherapy, but the R-IPI provides a simplified and more accurate prediction of outcome. The IPI factors can no longer be used to identify a group with less than a 50% chance of survival.

Overall Survival According to Revised IPI (R-IPI) Overall Survival According to Revised IPI (R-IPI)
Pilot Study of Lovastatin in Patients with Myelodysplastic Syndrome (MDS) Receiving Arsenic Trioxide (Trisenox®) Therapy.
Authors: Stroh Ann , Carter Thomas H. , Hohl Raymond J. , Zehr Pamela , Ritchie Justine M.
Published: April 2020

Statins have been shown to impair processing of RAS proteins, which are signaling molecules mutated in some patients with MDS, and the addition of lovastatin to myeloid cell lines exposed to arsenic trioxide in vitro has suggested enhanced cytotoxicity. To address the hypothesis that statins may modulate the therapeutic effect of arsenic, we are conducting a pilot study for MDS treated by arsenic trioxide combined with lovastatin. Objectives are to estimate the response rate, and secondarily to describe safety. Patients with MDS by FAB diagnostic criteria are treated with arsenic trioxide 0.25 mg/kg IV twice weekly for 16 weeks, after initial dosing for 5 consecutive days in the first week. Lovastatin is given at 20 mg/day for 2 weeks, then 40 mg/day for 2 weeks, and finally 80 mg/day for 12 weeks. Toxicity is monitored continuously and reviewed by a local Data and Safety Monitoring Committee. Stopping rules include serious adverse events (SAE) definitions for evidence of myopathy (symptoms or CPK >2x upper limit of normal) or hepatocellular injury (transaminases >3x upper limit of normal). Nine patients have been enrolled to date, with sufficient follow up to assess efficacy and safety. Five of these 9 patients completed the planned 16-week regimen. One subject was removed from study because of a rapid rise in blast count before treatment, based on marrow exams in a brief interval between screening and enrollment. Three patients had study drugs stopped per the above stringent SAE definitions, for elevations of CPK in 1 and transaminases in 2 subjects -- these abnormalities were asymptomatic and resolved. Toxicities were otherwise tolerable and as expected, including QTc prolongation in 1 patient, which resolved after withholding arsenic trioxide for several days and drug resumption was uneventful, as well as grade <=2 fatigue, nausea and insomnia. Responses are assessed by IWG criteria. One patient had a minor erythroid response with need for packed red blood cell transfusion decreased by 50%. One patient had a minor platelet response documented as a net increase greater than 10,000/mm3 but less than 30,000/mm3. One patient showed progression of disease to AML before completing the trial. Remaining patients showed stable disease; one showed marked but non-sustained improvement in counts and suffered necrotizing faciitis after completing the study as a complication of a bone marrow biopsy. Preliminary findings in an additional patient after 8 weeks of therapy suggest an early response in multiple lineages and freedom from red cell transfusion. Tentative conclusions are that this combination of a statin and arsenic trioxide has activity in MDS, and toxicities are tolerable.
Enhanced HDAC (Histone Deacetylase) Enzyme Activities in Blood CD 34+ Cells in Patients with Agnogenic Myeloid Metaplasia Are Not Related to Jak 2 Mutation.
Authors: Wang Jen-Chin , Boghossian Nubar , Chang Tsong S. , Novetsky Allan D. , Astrow Allan
Published: April 2020

We have previously reported that HDAC enzyme activities were elevated in patients with agnogenic myeloid metaplasia (AMM) as compared with patients with other myeloproliferative disorders including essential thrombocytosis (ET) and polycythemia vera (PV) and normal volunteer controls (Exp hematol33:1052005). Jak 2 mutations were found in 35–50% patients with AMM. The present study was designed to see if the elevated HDAC activities in AMM are related to JAK 2 mutations. 9 patients of AMM, and 17 of other MPD, and 10 normal volunteer controls were studied. Detection of Jak 2 mutation was using Bsa X1 digestion of the PCR product (Lancet365: 1054, 2005). 4 patients with AMM were found to have elevated HDAC enzyme activities (defined as more than 2 SD of controls) and only one patient was found to have an heterozygous mutation of Jak2 mutation.. We conclude that enhanced HDAC activities in AMM are not related to the Jak-2 mutation and enhanced HDAC activities may represent another mechanism than JAK2 mutations in the pathogenesis of AMM.
Validation of JAK2 and New Clinical Criteria for the Diagnosis of Polycythemia Vera (PV).
Authors: Silver Richard T. , Jones Amy V. , Feldman Eric J. , Roboz Gail J. , Ritchie Ellen K. , Cross Nicholas C.P.
Published: April 2020

Although the pervasive presence of JAK2 in patients (pts) with PV has been noted in at least 7 published studies, its frequency has ranged from 66 to 97%. Although part of the reason for this wide range is false negative results due to the low sensitivity of sequence analysis, varying criteria used for the clinical diagnosis of PV are also likely to be an important factor. In more than 300 studies, the criteria of the Polycythemia Vera Study Group has been used, although many authors, including ourselves, have commented on their inadequacy. Alternative classifications have been the subject of debate mostly centered about using the rbc-Cr51 mass. Our criteria included the use of CR51 labeled red cells to establish first, an increase in red blood cell volume and second, I125 to establish an increase in plasma volume. Exception was made for men with a HCT ≥60%, and women ≥56%. If hypervolemic, the major causes of secondary polycythemia were excluded: congenital polycythemias and those secondary to impaired oxygenation, or benign or malignant tumors. This implied a serum erythropoietin level of ≤5u/ml. The aforementioned and 3 of the following 5 were required: splenomegaly, WBC, ≥12,000/ul, platelet count ≥600,000/ul, abnormal marrow histology. We used them to clinically diagnose 61 patients subsequently undergoing therapeutic trials whose blood was analyzed for the JAK2 V617F mutation by allele specific PCR from 679 patients with myeloproliferative diseases, including normal controls. Of 61 patients, with the phenotypic characteristics of p. vera, 59 (97%) were positive for the JAK2 mutation. The 2 remaining patients, although phenotypically characteristic of PV, were negative on repeat examinations. One pt was newly diagnosed and untreated and the other had a 6 year remission after rIFNα followed by a 1 year remission on imatinib. We conclude that determination of JAK2 is consistent with PV in virtually all cases clinically diagnosed appropriately. Our data, to be presented, indicate that the combination of increased red cell mass, demonstration of JAK2 and increased platelet count establish the diagnosis of PV in virtually all cases.