bluuebunny/crossref_metadata_2025 huggingface dataset:
Anti-CD45 Mediated Cytolysis of Human T-Cell Lymphoma Cells.
Authors: Wulf Gerald G. , Boehnke Anita , Glass Bertram , Truemper Lorenz
Published: April 2020

Anti-CD45 mediated cytoreduction is an effective means for T-cell depletion in rodents and humans. In man, the CD45-specific rat monoclonal antibodies YTH24 and YTH54 are IgG2b subclass, exert a predominantly complement-dependent cytolytic activity against normal T-lymphocytes, and have been safely given to patients as part of conditioning therapies for allogeneic stem cell transplantation. The efficacy of such antibodies against human lymphoma is unknown. Therefore, we evaluated the cytolytic activity of YTH24 and YTH54 by complement-dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), as well as by direct apoptotic and antiproliferative effects, against a panel of Hodgkin disease (HD) and non-Hodgkin lymphoma (NHL) cell lines, and against primary specimens. Significant CDC activity (>50% cytolysis) of the antibodies YTH54 and YTH24 was observed against three of five T-cell lymphoma lines, but against only one of nine B-cell lymphoma lines and none of four HD cell lines. The combination of YTH54 and YTH24 induced ADCC in all T-cell lymphoma cell lines and three primary leukemic T-cell lymphoma specimens, but were ineffective in B-cell lymphoma and HD cell lines.There were only minor effects of either antibody or the combination on lymphoma cell apoptosis or cell cycle arrest. In summary, anti-CD45 mediated CDC and ADCC via the antibodies YTH24 and YTH54 are primarily effective against lymphoma cells with T-cell phenotype, and may be an immunotherapeutic tool for the treatment of human T-cell lymphoma.
A Phase II Study of Xcellerated T Cells™ in Patients with Relapsed or Refractory Indolent Non-Hodgkin’s Lymphoma (NHL).
Authors: Bartlett Nancy L. , Gribben John G. , Boccia Ralph V. , Milder Michael S. , Aboulafia David M. , Castro Januario , Flinn Ian , Hayes-Lattin Brandon , Kaplan Lawrence , Lin Thomas , Lucas Jennifer B. , Maloney David G. , McLaughlin Peter , McSweeney Peter A. , Mohrbacher Ann , Redfern Charles , Steis Ronald , Stephenson Joe J. , Strair Roger , Berenson Ronald J. , Frohlich Mark W.
Published: April 2020

Background: T cells can be activated and expanded ex vivo using the Xcellerate™ Process, in which peripheral blood mononuclear cells (PBMC) are incubated with anti-CD3 and anti-CD28 antibody-coated magnetic beads (Xcyte™-Dynabeads®). In an ongoing trial of Xcellerated T Cells in subjects with chronic lymphocytic leukemia, marked and sustained reductions in lymphadenopathy and splenomegaly were observed (Wierda et al., ASCO 2004). Increases in neutrophil, platelet and NK cell counts were also documented. This study is designed to determine if similar effects can be observed in subjects with indolent non-Hodgkin’s lymphoma (NHL).

Methods: Subjects must have indolent NHL (follicular, small lymphocytic, marginal zone, or mantle cell lymphoma), have relapsed or refractory disease, and have received at least 1 but not more than 4 prior treatment regimens. PBMC are collected by leukapheresis for the Xcellerate Process, and subjects subsequently receive two infusions of 20–60 x 109 Xcellerated T Cells separated by 6–8 weeks. Approximately 40 subjects will be treated.

Results: Seven subjects have been enrolled and Xcellerated T Cells have been manufactured in 5 subjects to date. T cells expanded 181.8 ± 88.5 fold and the final product was >99.0 ± 0.0% T cells (mean + SD). One subject with small lymphocytic lymphoma has been treated with two infusions of 38.6 x 109 Xcellerated T Cells. There have been no serious adverse events to date. Following the first treatment, the lymphocyte count increased from 1.8 x 109/L to 2.9 x 109/L on Day 28. The neutrophil count also increased from 2.9 x 109/L to 5.5 x 109/L six weeks following infusion. The subject had a significant reduction in cervical lymphadenopathy and a slight decrease in bulky mesenteric lymphadenopathy six weeks following the first infusion.

Conclusions: Xcellerated T Cells can be manufactured in subjects with indolent NHL. Treatment leads to significant increases in T cell and neutrophil counts. Preliminary data suggest a reduction in peripheral lymphadenopathy. Data on additional subjects will be presented.
Naïve T Cell Level and TCR Vβ Repertoire Usage in Patients with Chronic Myelogenous Leukemia.
Authors: Li Yangqiu , Geng Suxia , Yang Lijian , Chen Shaohua , Yin Qingsong
Published: April 2020

During early T lymphopoiesis, rearrangement of the V, D and J segments of the TCR genes result in deletion of the intervening chromosomal DNA and formation of circular excision circles, the so-called signal joint T-cell receptor rearrangement excision circles (sjTRECs, TRECs). TRECs are assumed to have a high stability, and can not multiply and consequently are diluted during T cell proliferation. It has been used to evaluate thymic function in T-cell immune reconstruction after treatment in HTLV-I-infected or stem cell transplantation. Defects of cellular immunity, however, may also play a role in hematologic malignancies. On the other hand, examination of T cell receptor (TCR) gene repertoire is important to analysis the immune status of the patients with malignant neoplasms, because clonal expansion of T cells permit the identification of specific antigen response of T cells. Little is known about the feature of T-cell immune state in CML. In order to evaluate the thymic recent output function and the expansion feature of TCR V beta subfamily T cells in patients with chronic myelogenous leukemia (CML in chronic phase), TRECs level and TCR V beta repertoire usage and clonality were analyzed. Quantitative detection of TRECs in DNA of peripheral blood mononuclear cells from 20 cases with CML, purified CD4+ or CD8+ cells from 3 cases with CML were preformed by real-time PCR (TaqMan) analysis. And the TRECs-number was related to the number of T-cells by determination of the number of CD3-positive cells. The expression and cloanlity analysis of TCR V beta repertoire were detected by RT-PCR and genescan technique in PBMC from the 14 out of 20 patients. 9 normal individuals served as controls. A dramatic reduction of TRECs values in patients with CML was showed. The mean value of TRECs was 0.06±0.16 copies /1000 T cells in CML patients and 6.84±4.71 copies /1000 T cells in normal individuals, respectively (p<0.01). In 16 out of 20 CML cases no TRECs copies could be detected in peripheral blood, and lower value of TRECs could be identified in sorted CD4+ or CD8+ cells. The expression of 1–12 V beta subfamilies was found in samples from 14 patients. Clonal expanded T cells from 13 cases could be identified in some V beta subfamilies, which were preferentially used in V beta 3, V beta 10, V beta 19, V beta 21 and V beta 22 subfamilies. In conclusion, this is, to our knowledge, the first description of TRECs level in CML patients. These results showed a prominent reduction of TREC levels in CML. The predominant usage and clonal expansion of TCR Vβ subfamily T cells could be identified in CML patients, indicating the host could have the ability for specific immune response to leukemia associated antigen, in despite of T cell immunodeficiency was showed in patients.
Successful Peripheral Blood Stem Cell (PBSC) Mobilisation with G-CSF in Patients with Chronic Myeloid Leukaemia Achieving Complete Cytogenetic Remission with Imatinib.
Authors: Carella A.M. , Corsetti Maria T. , Bodenizza Carlo , Iuliano Francesco , Pucci Giulia , Jacopino Pasquale , Martino Bruno , Brugiatelli Maura , Ferrara Felicetto
Published: April 2020

The clinical studies have demonstrated the efficacy of Imatinib in the induction of hematological remission, cytogenetic remission (CCR) and molecular reduction of the bcl-abl transcript as shown by RT-qPCR. Unfortunately, the optimal long-term management of patients who achieve CCR after Imatinib is unknown. It is unclear that Imatinib alone will prove to be curative and initial responders may eventually lose Imatinib responsiveness. Therefore it may be prudent to collect autologous PBSC in CCR patients treated with Imatinib with low levels of detectable leukemia analyzed by molecular tests. We evaluated G-CSF mobilisation of PBSC in 18 patients who have achieved CCR with Imatinib. Our data demonstrated that the target CD34+ cell yields of ≥ 2.0x106/kg was attained with G-CSF at the dose of 10 mg/kg/day in 4/8 (50%) patients during uninterrupted Imatinib therapy and in 8/10 (80%) when Imatinib was temporarily interrupted. Three patients (37%) in the first group and 7 patients (70%) in the second group achieved >1x106/kg CD34+ cell yield per apheresis. Twelve patients were evaluated on PBSC for bcr-abl by RT-qPCR. Three patients were negative and in the other 9 patients, a median of 0.20 (range, 0.02–8.6) remained detectable. These data compared favourably with a median of 0.04 (range, 0.02 – 0.86) of all measurements taken before mobilisation. There was no impact of G-CSF mobilisation on the CML as measured by cytogenetic and serial blood bcr-abl levels. In conclusion, PBSC mobilisation with Imatinib and G-CSF in CCR patients is feasible, CD34+ cell yield is significantly better with temporary withheld of Imatinib, G-CSF did not preferentially mobilize leukemic progenitors and leukemic burden did not show significant change in the months following G-CSF mobilisation.
Imatinib Therapy for Chronic Myeloid Leukemia Patients Who Relapse after Allogeneic Stem Cell Transplantation: A Molecular Analysis.
Authors: Palandri Francesca , Martinelli Giovanni , Amabile Marilina , Bonifazi Francesca , Bandini Giuseppe , Benedetti Fabio , Usala Emilio , Angelucci Emanuele , Tiribelli Mario , Fanin Renato , Sorio Marco , Arpinati Mario , Testoni Nicoletta , Falcioni Sadia , Giannini Maria Benedetta , Poerio Angela , Soverini Simona , Pane Fabrizio , Rosti Gianantonio , Tura Sante , Baccarani Michele
Published: April 2020

We analyzed clinical and molecular follow-up of 16 patients with chronic myeloid leukemia (CML) relapsing after allogeneic stem cell transplantation (SCT).

The median interval between allogeneic SCT and relapse was 26 months (7–162). Two patients had failed treatment with donor lymphocyte infusions prior to Imatinib; four patients had received therapy with IFN alpha. All patients were treated with Imatinib (400 or 600 mg/daily). One patient had received Imatinib before allogeneic SCT. The overall complete hematological (CHR) and cytogenetic responses (CCyR) were 100% for all patients either relapsed in CP or AP. All patients achieved complete molecular response (CMR), intended as 3 logs reduction of BCR-ABL/B2M within 18 months; 8/16 patients obtained a CMR within three months, independently of the phase of the disease at of relapse. Median follow-up after start of Imatinib therapy was 24 months (range 6–36). Chimerism status evaluated in 9 patients showed conversion to full donor chimerism after therapy in all but one of them. Imatinib has significant activity against CML in relapse after allogeneic bone marrow transplantation with durable cytogenetic and molecular remissions obtainable in all patients.
Imatinib Achieves High Rates of Complete Citogenetic Remission (CCR) in CML Patients Relapsed after Autografting anf IFN-α Therapy.
Authors: Corsetti Maria T. , Beltrami Germana , Carella A.M.
Published: April 2020

Background: Imatinib, a selective inhibitor of the bcr-abl tyrosine kinase, induces CCR in the majority of patients with CML in first chronic phase (CP). In the present study we addressed the question if such achievement of CCR can occur also in patients relapsed after autografting and IFN-α therapy.

Methods: from 1991 to 2001, 50 patients with early chronic phase were treated with hydroxyurea; subsequently, when WBC count was less than 10x109/L, the patients were treated with idarubicin containing regimen (ICE/mini-ICE protocols), mobilization of diploid or prevalently diploid peripheral blood stem cells (PBSC) with G-CSF during the early phase of recovery after chemotherapy and high-dose therapy followed by autologous diploid PBSC previously collected and cryopreserved. After engraftment, the patients received maintenance therapy with IFN-α (3MU/three times weekly). From september 2000, Imatinib became available for CML patients relapsed after IFN-α treatment. From that time all patients who relapsed after autografting and IFN-α received rescue therapy with Imatinib at 400 mg/daily for CP (Novartis protocol CSTI-106) and 600 mg/daily for accelerated phase (AP) (Novartis protocol-Italian Cooperative Study Group CSTI-003). Imatinib was given to 22 patients cytogenetically relapsed in CP and to 1 patient evoluted in AP.

Results: At the time of analysis (July 31, 2004), the median treatment duration with Imatinib was 32 months (range, 1 to 47). Four patients stopped Imatinib between 9 and 12 months for no response/progression. The patient in AP died of heart failure one month after starting Imatinib; since this patient have had previous history of cardiac failure, it is not clear if this event should be correlated with Imatinib therapy; two other patients died for progression. Imatinib induced major cytogenetic responses in 16/23 patients (70%) of whom 14/23 achieving CCR (61%). At a median follow up of 36 months the PFS and OS were 78% and 87%, respectively. Grade ≥III hematological toxicity occurred in 5 patients; this toxicity resolved with temporary discontinuation of the treatment.

Conclusion: Imatinib is an effective therapy for CP-CML patients who relapsed after autografting and IFN-α. The high rate of sustained haematological and cytogenetic responses achieved with Imatinib were associated to a favourable and manageable toxicity profile.
The NK and NKT Cell Compartment Is Suppressed in CML Patients before and after Imatinib Treatment.
Authors: Chen Christiane I.-U. , Maecker Holden , Lee Peter P.
Published: April 2020

In CML patients, lymphocyte subsets have been shown to be disturbed. NK cells as well as CD8+ T cells have been shown to be significantly lower in patients with relapse after BMT than without relapse (Jiang et al., 1996). After successful treatment (IFN- α and BMT/SCT), MHC-restricted CD8+ T cells as well as MHC-unrestricted cytotoxic NK and lymphokine activated killer (LAK) cells play an important role in controlling the disease (Meseri et al., 1993, de Castro et al., 2003, Molldrem et al., 2000), which can lead not only to hematological, but also cytogenetic responses. The development of imatinib mesylate, a selective inhibitor of the bcr/abl tyrosine kinase, as a first-line therapy enormously enhanced treatment options for CML patients, and has led to hematologic, and occasionally cytogenetic, responses at various stages of disease. To explore, if CML patients, who develop clinical responses with imatinib mesylate treatment, have normalization of their lymphocyte subsets with restoration of CD8+ and NK cells, we studied peripheral blood T cell, B cell, and NK cell subpopulations in patients in chronic or accelerated phase before and after treatment with imatinib mesylate. In 5 CML patients, 8-color flow cytometry was applied to investigate the lymphocyte subsets using anti-CD3, anti-CD4, anti-CD8, anti-CD19, anti-CD56, anti-CD25, anti-CD27, and anti-CD45RA antibodies. 5 healthy donors served as controls. There was no significant difference in CD19+ B cells or CD3+ T cells as well as its CD4+ and CD8+ subsets between CML patients and healthy controls. T regulatory cells (CD4+CD25+) were also similar in patients in comparison to the healthy controls (0.6% – 3% vs 2.3%). In contrast, CD3–CD56+ NK cells (4% vs 14%, p<0.05) as well as NK T cells (CD3+CD56+ 1% vs 14%, p<0.02; CD8+CD56+ 2% vs 17%, p<0.03) were profoundly decreased in CML patients compared to the healthy controls prior to treatment with imatinib mesylate. Importantly, these abnormalities persisted even during treatment and following hematological responses (range 10 months – 35 months) (CD3-CD56+ 4–8%, CD3+CD56+ 0.5–5.5%, CD8+CD56+ 2.7–7%). On further analysis, these NK and NKT cells were mostly effector (CD45RA+CD27−, 50–80%) or memory (CD45RA-CD27+, 10–30%) cells. These results suggest, that CML not only disturbs the myeloid and the lymphoid compartment before treatment, but persistently impairs the MHC-unrestricted cytotoxic NK cells and NKT cells after treatment with imatinib mesylate, even in patients in hematological (and cytogenetic) remission with normal white cell counts (2.0 – 8.0/nl). Further investigations are required to better understand the mechanism of the prolonged and specific NK and NKT cell suppression in imatinib mesylate treated patients, and its impact on the course of the disease.
Phase II Pilot Study of Imatinib Mesylate Combined with Induction Chemotherapy in Blast-Phase CML and Ph+ ALL.
Authors: Lickliter Jason , Arthur Chris , D’Rozario James , Hui Chi-hung , Szer Jeffrey , Taylor Kerry , Watson Anne-Marie , McCarron Jennifer , Lynch Kevin , Bradstock Ken
Published: April 2020

CML in blast crisis (BC) and Philadelphia chromosome-positive (Ph+) ALL are incurable with conventional chemotherapy and in most cases respond only transiently to the Bcr-Abl kinase inhibitor imatinib mesylate. We hypothesized that pretreatment with imatinib would overcome inherent drug resistance to induction chemotherapy mediated by Bcr-Abl, and evaluated the combination in 17 patients with Ph+ acute leukemias (5 CML in BC, 6 de-novo Ph+ ALL, 6 relapsed Ph+ ALL). All patients received a 1-week prephase of imatinib alone at 600 mg daily. In CML in BC and relapsed Ph+ ALL, imatinib was continued concomitantly with induction chemotherapy (standard-dose idarubicin/ara-C with vincristine/prednisone added for lymphoid leukemias) for an additional week. Imatinib was then ceased, but recommenced as a single agent after blood-count recovery. Patients with de-novo Ph+ ALL received the imatinib prephase and then commenced the French LALA94 protocol without concomitant imatinib. Neutrophils recovered to ≥ 1.0 x 109/L at a median of day 28 (range 21–56) of induction chemotherapy. During induction, 1 patient died from sepsis and multi-organ failure, and 1 patient died on day 7 from CNS leukemia.

Subsequently, 1 patient died on day 37 from massive hemoptysis and 1 patient developed pulmonary fibrosis and died on day 144. Among 14 evaluable patients, combined imatinib and chemotherapy induction resulted in 9 (64%) complete hematological responses and there were 5 complete and 3 major cytogenetic responses. Notably, 2 out of 3 patients with CML in myeloid BC achieved major cytogenetic responses. The in-vivo effects of imatinib on Bcr-Abl activity were monitored in leukemic blasts during the imatinib-only prephase. Western blots revealed a decrease in CrkL phosphorylation to < 25% of the pretreatment level in peripheral-blood samples from 5 out of 6 evaluable cases and in marrow samples from 1 of 2 evaluable cases. Bcl-xL expression levels revealed no consistent in-vivo responses to imatinib. However, all 7 cases evaluable for Bcl-2 protein showed strong expression levels, which were unaffected by imatinib in 5 cases and decreased in 2 cases. Imatinib potently inhibited Bcr-Abl activity in vivo in Ph+ acute leukemic blasts and its combination with induction chemotherapy was tolerable and shows promising activity.
Monitoring of BCR-ABL Transcirpt Levels after Discontinuation of Imatinib Therapy in Chronic Myelogenous Leukemia Patients Achieving Complete Cytogenetic Response.
Authors: Kim Yoo-Jin , Kim Dong-Wook , Lee Seok , Eom Ki-Seong , Min Chang-Ki , Kim Hee-Je , Lee Jong-Wook , Min Woo-Sung , Kim Chun-choo
Published: April 2020

Background Imatinib induces a high complete cytogenetic response (CCR) rate in chronic myelogenous leukemia (CML). None of the studies addressed the important question of how long imatinib treatment should be continued. To answer this question, we tried to assess the minimal residual disease after cessation of imatinib in patients achieving CCR.

Methods A total of 23 patients were enrolled and their disease status at the start of imatinib was chronic phase in 7, accelerated phase in 3, blast crisis in 1, and post-transplant relapse in 12 patients. All the patients were in CCR when imatinib was discontinued and BCR-ABL transcript was undetectable in 9 of them. Minimal residual disease was monitored using nested RT-PCR, real-time quantitative RT-PCR, and conventional cytogenetics.

Findings Duration of imatinb treatment and CCR before cessation of imatnib was a median of 13 months (range, 6–25 months) and 10 months (range, 1–22 months), respectively. After cessation of imatinib treatment, cytogenetic relapse was observed in 12 (53%) of 23 patients and hematological relapse was followed in 8 (67%) of them. Extramedullary relapse was documented in 1 patient. Prior allogenetic transplantation, molecular remission at the time of cessation, and time to CCR were potential variables affecting probability of cytogenetic relapse. Normalized BCR-ABL transcript level progressively increased after cessation of imatinib in all but 3 patients who had been treated with allogeneic transplantation. Restart of imatinib induced down-regulation of normalized BCR-ABL transcript level in 5 patients with available data.

Interpretation Imatnib should be maintained in the most of patients who achieve a short duration of CCR as in the cases with interferon treatment. However, this study suggests that it seems possible to stop imatinib treatment in the minority of CML patients who was treated with allogeneic transplantation.
Quality of Life and Economic Impact of Red Blood Cell (RBC) Transfusions on Patients with Myelodysplastic Syndromes (MDS).
Authors: Brechignac S. , Hellstrom-Lindberg E. , Bowen D. T. , DeWitte T. M. , Cazzola M. , Fenaux P.
Published: April 2020

Background: Supportive care with blood product transfusions is the primary management strategy for the majority of patients with MDS. Approximately 80% of MDS patients are anemic at the time of presentation and more than 40% require regular RBC transfusions at some stage of disease, while platelet transfusions are less often required.

Methods: In an effort to systematically study quality of life and economic cost associated with transfusion dependency (especially RBC transfusions), The MDS Foundation has disseminated a practices and treatment survey to its Centers of Excellence and is also accumulating transfusion data. Retrospective and prospective data collected include hematologic parameters defining transfusion need; percentage (%) of MDS patients requiring transfusion; % of transfusion-dependent MDS patients by subtype and International Prognostic Scoring System (IPSS) risk group; per patient frequency of transfusions; % of patients requiring iron chelation therapy.

Results: A total of 30 Centers have replied to the survey to date, and responses reveal that a substantial proportion of MDS patients receive multiple RBC transfusions with most of these patients needing chelation therapy with desferoxamine (generally subcutaneous administration, 4-times weekly): Table 1.

In addition, detailed data are available from 4 European Centers that have provided transfusion records from randomly selected multiply-transfused MDS patients: 38 patients (median age: 73) received a median of 42 transfusions over the last 24 months (range: 11–207). The average per transfusion costs calculated from estimates provided by the 4 European centers is 436 euros or $ 526 ($1 US dollar = 0.83 euros), where the per transfusion cost includes 2 filtered red blood cell units, blood collection, administrative costs, and staff time, resulting in a median per patient cost over the last 24 months of 11,118 euros (range: 5668–21,800 euros). This does not include the cost of chelation therapy (300 euros/month for desferioxamine SC) and indirect costs (e.g., time spent at transfusion facility, travel time for patient to facility, travel and wait time for private caretaker or family member).

Conclusion: Preliminary data analysis from the ongoing retrospective study suggests that the transfusion burden to MDS patients and to society, in terms of quality of life and cost, is much greater than generally appreciated. Updated data of this study will be presented.

Table 1: RBC Transfusion-dependent MDS patients

Mean % IPSS low risk 39 IPSS intermediate-1 risk 50 IPSS intermediate-2 risk 63 IPSS high risk 79 Iron chelation therapy 28
Treatment of Del(5q) Myelodysplastic Syndrome with All-Trans-Retinoic Acid and Tocopherol-α.
Authors: Giagounidis Aristoteles A.N. , Haase Sabine , Germing Ulrich , Schoch Claudia , Schlegelberger Brigitte , Wilkens Ludwig , Büsche Guntram , Kreipe Hans H. , Aivado Manuel , Lübbert Michael , Ganser Arnold , Aul Carlo
Published: April 2020

All-trans-retinoic acid (ATRA) alone or in combination with cytokines and vitamins has shown erythroid remitting capacities in low-grade myelodysplastic syndromes (MDS). We performed a phase II study on 29 patients with MDS and isolated del(5q) including bands 5q31–5q33 to determine the efficacy and safety of ATRA in combination with tocopherol-α. All patients had low/intermediate-1 risk MDS according to the international prognostic scoring system. Inclusion criteria were isolated del(5q), medullary blast count of <10%, hemoglobin <10g/dl or transfusion dependent anemia, and ECOG performance score <3. Included were 18 women and 11 men. Median age of patients was 64 years. FAB subtypes were 25 refractory anemias, 1 refractory anemia with ring sideroblasts, 3 refractory anemias with excess of blasts. Median MDS duration before ATRA treatmnet was 31.7 months. Median hemoglobin level was 8.4 g/dl at study entry. Patients received 45 mg/m² ATRA day 1 to 90, and 90 mg/m² day 91 to 180. Tocopherol dosage was 600 IU three times daily. 24 patients completed dose level I, 12 patients dose level II. 86% of patients experienced side effects. 13 interrupted study drug treatment due to adverse events: Skin reactions (NCI grade 3), conjunctivitis (grade 3), headache and joint pain (grade 3) were the worst recorded. Other adverse events included hair loss, CNS symptoms, cheilitis, stomatitis, creatinine elevation, hypertriglyceridemia, hypercholesterinemia, nausea and vomiting, and elevation of liver function tests. 3 patients were admitted to hospital for side effects and these were therefore qualified as serious. Response criteria were qualified as recommended by the international working group for MDS. One patient (3%) achieved a major erythroid response resulting in transfusion independence throughout the study. He became transfusion dependent again within one month after study termination. 4 patients (14%) achieved a minor erythroid response with reduction of >50% of transfusion needs. No patient had a cytogenetic response. Responders did not show a reduction in del(5q) as determined in interphase fluorescence in-situ hybridization. There was no significant improvement in quality of life in responding patients as measured by the EORTC quality of life questionnaire. We conclude that the combination of ATRA and tocopherol-α is not recommended for the treatment of del(5q) MDS.
The Expression of Platelet Membrane Glycoproteins in Patients with Myelodysplastic Syndrome (MDS).
Authors: Girtovitis Fotios I.F. , Papagiannis Vasilis , Klonizakis Ioannis , Papadopoulos Athanasios , Bougiouklis Dimitris , Boutou Afroditi A.K. , Makris Pantelis P.E.M.
Published: April 2020

The receptors of platelet membrane have a significant function. Most of them are glycoproteins and play a major role in the adhesion and aggregation of platelets, in a way that any change of their quantity or quality will lead to functional defects of the second.

Aim of this study is to determine the qualitative expression of platelet membrane glycoproteins among patients (pts) with MDS, as a reflection of the various defects that characterize those syndromes.

Materials and methods: 24 pts were studied, 15 male and 9 female with mean age 73.2 ± 13,4 years old, who suffered from all of the types of MDS (FAB classification).

Specifically, 9 patients (pts) presented RA, 2 pts RARS, 7 pts RAEB, 2 pts RAEB-t and 4 CMML. All of the pts, as well as 16 normal subjects (8 male and 8 female with 68,4±9.7 old age) who were used as controls, were investigated using flow cytometry and certain monoclonal antibodies(CD42a and CD42b for glycoproteins GPIb-V-IX, CD41 and CD61 for glycoproteins GPIIb-IIIa and CD62P for P-selectin). The statistical method used was the comparison of mean values by t-test.

Conclusions: As shown in the table above, GPIb IX-V (CD42b), P selectin and GpIIb are reduced in all of the categories of pts. As long as GpIIIa is concerned, it is reduced only among pts with RARS and RAEB. On the contrary, CD42a (GpIb) does not seem to have been affected by these hemopoietic disturbances. It is possible that the reduction of those receptors a) reflects changes in the normal production of platelets and b) explains functional disorders of platelets.

n CD42a % CD42b % CD41 % CD61 % CD62 % RA 9 97.6±3,29 49.5±25,2 60±32,4 91.8±17,2 8.2±7,4 RARS 2 80.5±17,5 52.8±42,8 36±35,8 72±26 6.7±3,3 RAEB 7 89.4±14,1 58.9±29,9 58.8±29 77.6±28,2 6.3±4,5 CMML 4 97.3±2,08 37,8±20,7 70.3±25,7 95.8±2,7 2±0,5 RAEB-t 2 95.3±2,15 32.8±4,95 44.1±9,15 93.9±1,85 7.2±6,7 Total 24 92±20,1 46.3 ±27,9 53.8±29,8 86.2±21,8 6±6 Controls 16 95.9±3,4 78.23±17,5 85.5±8,5 95.36±3,01 14.1±10,6
Prolonged Administration of Arsenic Trioxide (Trisenox®) for Patients with Myelodysplastic Syndromes (MDS) and Chronic Myelomonocytic Leukemia (CMML) at MD Anderson Cancer Center: A Phase II Study.
Authors: Beran Miloslav , Verstovsek Srdan , Kornblau Steven M. , Estey Elihu H. , Giles Francis , Garcia-Manero Guillermo , Kantarjian Hagop M.
Published: April 2020

The value of prolonged administration of arsenic trioxide (ATO) was examined in patients (pts) with MDS and CMML. ATO (0.25 mg/kg) was given intravenously over 1 hour daily for five days followed by 0.25 mg/kg twice weekly for 11 weeks. Pts were assessed at 4 weeks, at the end of the first course, and then monthly. Pts with stable disease were eligible for further courses. The study included 14 RBC transfusion-dependent MDS pts (6 RA/RARS, 8 RAEB; 12 IPSS risk Low/Int1, 2 Int2/High), and 3 CMML pts. Median age was 67 years (range 46–84). Six pts had a history of previous treatment other than supportive care. 16 pts were evaluable for toxicity and response. One pt received 3 courses, 3 received 2 courses, and 13 received 1 course. Hematologic responses (IWG criteria) were observed in 4 pts (25%) and 9 (6 MDS,3 CMML) had stable disease:

FAB Type of response Time to response Duration RA Minor erythroid 2 months 3 mo, RBC independence RAEB Major neutrophil and platelet 2 mo 3 mo RAEB BM blast decrease 18% to <5% 1 mo 14 mo, stable pancytopenia RAEB Hematologic/cytogenetic CR 1 mo 6 mo

Hematologic/cytogenetic CR was achieved in one pt: a 76 year old male, RAEB-2, Int 2, complex chromosomal abnormalities, who had previously failed thalidomide for 6 months and thalidomide + cyclosporin A for 3 months. Of the 3 CMML pts, one had transient reduction of absolute monocyte counts during each of 2 courses; all 3 had stable disease. In 24 courses there were 6 febrile episodes in 4 patients, requiring admission, all in pts with pre-existing neutropenia; 1 Grade 3 and 1 Grade 4 thrombocytopenia, 1 Grade 3 neutropenia, all requiring dose modification; 1 episode of accelerated functional cardiac rhythm; and 1 anemia-precipitated congestive heart failure, which was unlikely related to the drug. There were 22 episodes of Grade 1 or 2 drug-related toxicities. Four responses of 3–7+ mo. duration were observed, including 1 CR, suggesting activity in an as yet undefined subgroup of MDS pts. No benefit of prolonged treatment was documented. ATO was safe in this outpatient setting for elderly patients. Observed low toxicity and documented activity support future trials of ATO in combination with other agents in treatment of MDS.
2 Chlorodeoxyadenosine (2-CdA) and Cytarabine (ara-C) Combination Is Effective and Safe in Idiopathic Hypereosinophilic Syndrome (HES).
Authors: Jabbour Elias , Verstovsek Srdan , Gandhi Varsha , Cortes Jorge , Giles Francis , Koller Charles , O’Brien Susan , Plunkett William , Garcia-Manero Guillermo , Jackson Ellen , Kantarjian Hagop , Andreeff Michael
Published: April 2020

Hypereosinophilic syndrome (HES) is rare, disabling, and incurable disease. In a pilot study, the combination of 2-CdA and ara-C chemotherapy has been evaluated in 9 patients. The median time from diagnosis to therapy was 25 months. All patients had signs and symptoms of end-organ involvement. Seven patients received prior therapies. The median eosinophil count at presentation was 14 x109/L (0.3–110 x109/L) and the median percent eosinophilic bone marrow infiltration was 23% (6%–43%). Ara-C (1g/m2) was given intravenously over 2 hours at hours 0, 48, 72, 96, and 120; 2-CdA (12 mg/m2/day) was given as continuous infusion intravenously over 5 days starting at 24 hours. A second course at the same level was administered for patients who achieved complete remission (CR) and for patients who had >50% reduction in eosinophilic infiltration. Two patients studied for pharmacokinetics of ara-CTP (active metabolite of ara-C) in the circulating blood cells accumulated either similar or higher levels of ara-CTP after infusion of 2-CdA. Five patients (55%) achieved CR, one requiring 2 cycles. Elimination of eosinophilia was accompanied by the resolution of symptoms. The median disease free survival was 26 months; 1 patient remains disease free (after allogeneic transplantation) at 42 months. The median overall survival from diagnosis for CR patients was 44 mo. Treatment was well tolerated. Febrile neutropenia occurred in 28% of the 14 cycles given. The median time to recovery from neutropenia and thrombocytopenia was 17 and 39 days, respectively. There was one death related to disease progression. We conclude that 2-CdA + ara-C combination regimen is safe alternative therapy for HES. The addition of consolidation therapy or imatinib may improve the results.
Detailed Analysis of Apoptotic Responses to Cytokine Stimulation in B-Cell Chronic Lymphocytic Leukaemia.
Authors: Jewell Andrew P. , Smith Wendy , van Schie Rob , Porakishvili Nina , Lydyard Peter M.
Published: April 2020

Introduction B cell Chronic lymphocytic leukemia (BCLL) is characterized by the accumulation of monoclonal malignant CD5+ B cells that show resistance to apoptosis in vivo, but are susceptible to apoptosis when incubated in vitro. A number of cytokines have been reported to protect against apoptosis in vitro, but it is not clear whether the effects of these cytokines are mediated in similar ways. We have therefore studied the regulation of apoptosis in B-CLL cells in vitro, in response to various cytokines.

Methods B-CLL cells were purified and incubated with the following agents; Interferon-gamma, IL-4, IL-10, IL-13, TNF-alpha and GM-CSF, each at 3 different concentrations. Apoptosis was measured after 70 hours by Annexin/PI staining and analysed for early apoptosis (Annexin V positive), late apoptotic (Annexin V and PI positive), and necrotic (PI positive).

Results The percentage of live cells in unstimulated B-CLL cells was 42.4±2.3%. Co-incubation of B-CLL cells with IL-4 at 100ng/ml increased the percentage of live cells to 63.2±8.4%, while in the presence of interferon-gamma the percentage of live cells was 48.7±5.9%. In contrast, co-incubation with TNF-alpha reduced the percentage of live cells to 30.8±6.8%. IL-10, IL-13 and GM-CSF had no effect on B-CLL cells survival at any of the concentrations used.. No significant differences in the pattern of early/late apoptosis or necrosis were seen between the different cytokines used.

Figure Figure
Molecular Events Underlying the Induced Differentiation of CLL Cells Are Recapitulated in Cess B Cells.
Authors: Battle Traci E. , Frank David A.
Published: April 2020

Although much progress has been made in recent years in understanding the molecular pathogenesis of CLL, new therapeutic approaches are critical to improving patient outcome. One attractive approach is differentiation therapy, in which the malignant B cells are induced to mature into non-replicative cells with a shorter lifespan. We have shown previously that the natural product Bryostatin 1 induces differentiation of CLL cells through activation of the transcription factor STAT1. To identify genes activated by STAT1 that mediate Bryostatin 1-induced differentiation of CLL cells we used gene expression microarrays. Using this approach, we identified seven genes that were potential STAT1 target genes, including GADD45β, IFI16, caspase 3, topoisomerase I, and the known STAT1 target IRF-1. Given the limitations of genetically manipulating primary CLL cells, we evaluated whether B lymphocytic cell lines recapitulated the molecular and cellular differentiation responses to Bryostatin 1 that we observed in CLL cells. We found that Bryostatin 1 is a potent inducer of differentiation of CESS B lymphoblastoid cells, as measured by growth arrest, downregulation of MHC II, and IgG secretion. Bryostatin 1 also induced signaling events in CESS cells similar to those observed in CLL cells, including activation of STAT1 and induction of STAT1 target genes. These findings suggest that STAT1 and its targets may be important mediators of malignant B cell differentiation and that the CESS cell line can be used to further study the potential clinical application of differentiation therapy in CLL.
Correlative Study of ZAP-70 Gene Expression by Quantitative PCR and CD-38 Surface Marker in Patients with Chronic Lymphocytic Leukemia (CLL).
Authors: Larson Melissa L. , Shammo Jamile M. , Enschede Sari H. , Venugopal Parameswaran , Russell Daniel , Coon John S. , Gregory Stephanie A.
Published: April 2020

Patients with chronic lymphocytic leukemia have heterogeneous clinical courses. Many possible prognostic factors have been evaluated as a way to predict each patient’s future course. Two such prognostic factors are IgVH gene mutational status and CD38 expression. Recently, ZAP-70 expression has been found to correlate with IgVH gene mutation, but little is known about the correlation between ZAP-70 and CD 38 expression. We sought to evaluate the concordance between the two. METHODS: A procedure was developed at our institution to evaluate ZAP-70 transcripts by PCR. The results were then confirmed by flow cytometry. The blood or bone marrow samples of 14 patients diagnosed with CLL were analyzed by PCR for ZAP-70 transcripts and flow cytometry for ZAP-70 protein expression and CD38 expression. The results of the ZAP-70 transcripts and CD38 expression were then correlated to the clinical courses of the patients. RESULTS: Of the 14 patients analyzed, the majority had Rai Stage I disease at diagnosis (4 Stage 0, 8 Stage I, and 2 Stage II). There was a median follow-up of 28 months. The median number of treatments for these patients was one (range 0–6). Overall, the results for ZAP-70 and CD38 expression were concordant in eight with CD38 data unavailable for 2 patients. In the four patients with discordant results, 3 of them had high CD38 expression and low ZAP-70. However, patients with high ZAP-70 were more likely to require multiple treatments once treatment was indicated. Patients with high CD38 were also more likely to be treated. Cytogenetic data was available for 9 patients with the following results: 5 with normal cytogenetics, 1 with deletion of 11q, 1 with deletion of 13q, and 2 with deletion of 17p. Five of the nine patients had high ZAP-70 expression. CONCLUSIONS: In our institution, the results of ZAP-70 and CD38 expression were concordant 67% of the time. Longer follow-up is necessary and may further delineate the roles of cytogenetics, ZAP-70 expression, and CD38 expression on prognosis in CLL.
Immunoglobulin Heavy Chain (IgH) Gene Somatic Hypermutation Status Using Homoduplex-Separated RT-PCR Products.
Authors: Melan Melissa A. , Foon Kenneth A. , Marks Stanley M. , Kant Jeffrey A.
Published: April 2020

Assessment of IgH somatic hypermutation status has been shown to be a valuable indicator for judging the prognosis of patients with chronic lymphocytic leukemia (CLL). Our laboratory has developed a streamlined method to improve the rate of successful evaluation of IgH mutation status. Six individual PCR reactions are first performed using random hexamer-generated cDNA as template. These reactions have identical reaction parameters, use a common JH reverse primer and one of six VH class-specific forward primers within Framework 1. PCR products are separated on acrylamide or MetaPhor® agarose gels following formamide denaturation. In almost all cases, a single homoduplex band is resolved indicating a class-specific clonal product. The homoduplex band is excised from the gel, eluted and directly sequenced. Mutation analysis is performed using the NCBI Ig BLAST program with percent identity determined for the region from the beginning of CDR1 to the end of Framework 3. To date, less than 10% of cases analyzed have not yielded a clearcut clonal PCR product using this approach
The Effect of Subsequent Therapies in the Patients with Chronic Lymphocytic Leukemia Previously Treated with Cladribine and Prednisone or Chlorambucil and Prednisone in Randomized Studies.
Authors: Robak Tadeusz , Blonski Jerzy Z. , Kasznicki Marek , Gora-Tybor Joanna , Dmoszynska Anna , Wojtaszko Maria , Skotnicki Aleksander , Nowak Wieslaw , Hellmann Andrzej , Lewandowski Krzysztof , Zdziarska Barbara , Konopka Lech , Ceglarek Bernadeta , Dwilewicz-Trojaczek Jadwiga , Boguradzki Piotr , Kuliczkowski Kazimierz , Sulek Kazimierz , Warzocha Krzysztof
Published: April 2020

Previously, we reported a study on efficacy and toxicity of cladribine (2-CdA) + prednisone (P) compared with chlorambucil (Chl) + P in previously untreated patients with progressive or symptomatic chronic lymphocytic leukemia (CLL) in a randomized multicenter trial (Blood2000, 96, 2723). Of 229 patients enrolled, 126 received 2-CdA+P and 103 received Chl+P. Here, we present the long-term follow-up and the results of subsequent treatment in refractory or relapsed patients. Patients with non responsive (NR) disease or with progression after 3 courses of 2-CdA + P or Chl+P or who relapsed earlier then 12 months after completing the treatment were switched to an alternative arm. Patients who relapsed later than 12 months from first remission (late relapse) were retreated with the same schedule that induced previous response.

In 50 patients, non-responsive or with early relapse, treated originally with Chl+P who received 2-CdA+P as second line, complete response (CR) was achieved in 12 (24%) and overall response (OR) in 32 (64%). Twenty eight patients originally treated with 2-CdA+P received Chl+P and CR and OR was obtained in 1 (3,6%) and in 6 (21.4%), respectively (p=0.002 for both arms).

Thirty three patients with late relapse were retreated with 2-CdA+P and 19 patients were retreated with Chl+P. OR (CR) was 54.6% (6.1%) and 47.4% (15.8%), respectively (p=0.34).

The third line treatment was CHOP in both arms. Twenty seven patients from 2-CdA+P group and 37 patients from Chl+P group were treated with CHOP and only one patient responded.

Median survival was 3.3 and 3.75 years, respectively (p=0.63). Secondary neoplasms were observed in 8 (6.4%) patients treated with 2-CdA+P and in 4 (3.9%) treated with Chl+P as first line treatment (p=0.6). Death rates have been similar in patients treated originally with 2-CdA+P 82 (64.6%) and Chl+P 69 (66.4%) (p=0.8).

In conclusion, cladribine is significantly more effective as a second line treatment than chlorambucil. Both agents are similarly effective in retreatment of late relapsed patients. CHOP is of no value in patients previously treated with 2-CdA + P and with Chl + P. There is no significant difference in median survival of patients treated initially with Chl + P and 2-CdA + P.
Structural and Functional Characterization of the NHR2 and Runt Domains of AML1/ETO.
Authors: Cheney Matthew D. , Liu Yizhou , Zhou Yunpeng , Chruszcz Maksymilian , Laue Thomas M. , Minor Wladek , Bushweller John H. , Speck Nancy A.
Published: April 2020

AML1/ETO is the chimeric fusion protein resulting from the t(8;21) found in AML of the M2 subtype. It contains the N-terminal 177 amino acids of RUNX1 and virtually all (575aa) of ETO. The RUNX1 component includes the Runt domain, which mediates both DNA binding and heterodimerization with CBFβ, but lacks the more C-terminal sequences required for transactivation. AML1/ETO occupies RUNX target genes in vivo and is associated with a repressive chromatin structure characterized by reduced levels of acetylated histone H3. AML1/ETO is thought to repress transcription by recruiting a SMRT (N-CoR)/Sin3A/HDAC complex to chromatin via sequences in ETO. ETO is the human homologue of the Drosophila Nervy protein and shares 4 regions of homology with Nervy called Nervy Homology Regions (NHR) 1–4. Deletion studies have shown that three of the AML1/ETO domains essential for its repressive function are the Runt domain, NHR2, and NHR4. The NHR2 domain is a hydrophobic heptad repeat that mediates oligomerization of AML1/ETO, interaction with ETO family members, and also with mSin3A and HDACs. We recently solved an x-ray structure of the NHR2 domain and found it to be an alpha-helical tetramer. Based on this structure we have introduced amino acid substitutions into the NHR2 domain that disrupt tetramer formation but not AML1/ETO stability. These mutations impair the ability of AML1/ETO to inhibit the differentiation of GR-1+/Mac-1+ cells following retroviral transduction into primary mouse bone marrow cells, and also inhibit the serial replating ability of AML1/ETO expressing bone marrow cells in vitro. We additionally show that mutations reported by Amann et al. (Mol Cell Biol. 21, 6470, 2001) to disrupt mSin3A binding to NHR2 do not affect the biological activity of AML1/ETO in vitro. We also introduced mutations in the Runt domain of AML1/ETO that disrupt CBFβ binding by defined amounts (40-fold, 200-fold, 500-fold), and demonstrated that CBFβ binding by AML1/ETO is essential for its dominant negative activity. The latter results suggest that small molecules designed to selectively impair heterodimerization of AML1/ETO with CBFβ could potentially block AML1/ETO’s dominant negative activity.