bluuebunny/crossref_metadata_2025 huggingface dataset:
Little Evidence of the Protective Role of MTHFR Genetic Polymorfism in Acute Leukaemia and Down Syndrome in Brazil.
Authors: Zanrosso Crisiane W. , Emerenciano Mariana , Ramos Flavio , Figueiredo Alexandre , Magalhaes Isis Q. , Leite Edinalva , Cavalcanti Vanessa , Muniz Maria TC , Pombo-de-Oliveira Maria S.
Published: October 2019

Immunomolecular and genetic markers are great tools to understand the pathway of leukemogenesis. Because of acute leukemias (ALs) commonly arise as result of DNA damage and/or DNA translocations, the polymorphism of 5,10-methylenetetrahydrofolate reductase gene (MTHFR) have been associated to reduced enzyme activity and altered distribution of cellular folate metabolites. We have determined the prevalence of MTHFR mutations C677T and A1298C in 227 children with AL from different regions of Brazil and 279 region-matched subjects as the control group. PCR-RFLP method was applied using HinfI and MboII restriction digestion to determine the MTHFR 677 and 1298 respectively. Odds ratio (OR) and 95% confidence intervals (95%CI) were used as statistics standard methods. There were 202 acute lymphoblastic leukemias (ALL) and 65 acute myeloid leukemias (AML) from three different regions of Brazil. The frequencies of the 677CC, 677CT and 677TT genotype were 57%, 33%, 10% in the ALL cases; and 51%, 40%, 9% in the controls, respectively. Whereas the frequencies of MTHFR 1298AA, 1298AC and 1298CC were 52%, 40%, 8% in the ALL cases and 61%, 32%, 7% in the controls, respectively. Also, we analyzed separately a group of Down syndrome (DS) cases that include (i) DS with AL and, (ii) DS without AL. The frequencies of the 677CC, 677CT and 677TT genotype were 2,16%, 2,37%, 0% in the DS with AL cases, and 4,31%, 3,02%, 0,22% in the DS without AL, respectively. Whereas the frequencies of MTHFR 1298AA, 1298AC, 1298CC were 2,94%, 1,05%, 0,84% in the DS with AL and 4,83%, 2,10%, 0,63% in the DS without AL, respectively. There were no differences of MTHFR C677T frequencies in both groups in overall analysis.

However, the frequency of MTHFR 677 and MTHFR 1298 genotypes differ between cases and controls in different regions of Brazil (p<0.04). The overall analysis indicates that MTHFR C677T and A1298C may confer a protective effect against childhood acute leukemia only in distinct group of cases.

Region OD IC (95%) p South 0.41 0.045–3.59 0.69 Southeast 0.78 0.265–2.33 0.84 Northeast 1,02 0.385–2.65 0.69
Elevation of Normal B Cells in Children with ALL Can Predict Relapse.
Authors: Kapelushnik Joseph , Levi Itai , Yermoau Tikva , Moser Asher , Haj-Yhia Atamma , Shubinski Geora
Published: October 2019

Flow cytomerty analysis of bone marrow (BM) is performed routinely in children with acute leukemia in different phases of chemotherapy. The aim of flow cytometric monitoring is to measure the amount of residual blasts and to evaluate the efficacy of chemotherapy. Multiparametric flow cytometry may be also used to evaluate normal hematopoisis in BM of patients with acute leukemia in different phases of chemotherapy. The aim of this study was to study normal lymphopoiesis in patients with relapsed disease. Twenty-two pediatric patients with ALL were enrolled in the protocol of study. Patients were treated in accordance with BFM-98 protocol. Eighteen children were in complete remission during follow-up period, while 4 children relapsed. Three of the relapsed children had B-lineage ALL, and one had T-cell ALL. Using multiparamertic flow cytometry we measured normal pro-B cells (CD19+CD34+), immature B (CD19+CD10+CD20+ and CD19+CD10−CD20+) and mature B (CD19+CD10−CD20+) cells in BM of relapsed patients in different phases of chemotherapy. Amounts or different subsets of B cells were compared with reference values determined in patients with relapse-free disease. Reference values were expressed as mean ± confidence interval for each subset of B cells and for each phase of chemotherapy. Elevation of immature normal B-cells was determined in all 4 patients who relapsed.Three during chemotherapy. Increased amount of mature B cells was determined 3 mo before the diagnosis of relapse in the patient who relapsed after the end of chemotherapy.This event was not observed in patient who did not relapse We conclude that elevation of normal immature B cells in BM of patient with ALL can be a predictive marker for eventual relapse.
Sensitivity of Childhood Acute Lymphoblastic Leukemia (ALL) to Idarubicin Is Significantly Higher Than to Daunorubicin Treatment Based on Ex Vivo Apoptosis Induction.
Authors: Prokop Aram , Bagci Banu , Lingfeld Guenaelle , Badiali Lucia , Garbrecht Karin , Mietzsch Ulrike , Wieder Thomas , Daniel Peter T. , Henze Günter
Published: October 2019

Anthracyclines, especially daunorubicin, play a very important role in the treatment of acute lymphoblastic leukemia (ALL) and the relapsed ALL in childhood.

In the present study, primary lymphoblasts isolated from 65 children with de novo ALL (median: 5.8 years; range: 1.9 – 16.9 years) and relapsed ALL (median: 12.7 years; range: 1.3 – 17.9 years) were treated with daunorubicin (10 mmol/l) or idarubicin (2 mmol/l) in vitro. We could show that both anthracylines induce apoptosis, as evidenced by measurement of genomic DNA fragmentation. Interestingly, daunorubicin only induced modest apoptosis, whereas idarubicin displayed a significantly stronger apoptosis inducing effect. Furthermore the treatment of daunorubicin-resistant lymphoblasts with idarubicin resulted in good response in most of the resistant cell populations.

Out of the 65 patients analysed in this study 23 were female (13 de novo ALL, 10 relapsed ALL) and 42 were male (29 de novo ALL, 13 relapsed ALL). Primary lymphoblasts were obtained by bone marrow aspiration and separated by centrifugation over Ficoll. Within these cell populations following immunologic subgroups were found: 35 c-ALL, 10 pre-B-ALL, 7 pro-B-ALL, 10 T-ALL and 3 pre-T-ALL.

Daunorubicin induced apoptosis in 33 out of 65 lymphoblast populations (response rate 50.8 %). Nevertheless, a far higher response rate was observed for idarubicin with 59/65 (90,8 %) (p < 0.008), if response is defined as apoptosis induction higher than 1 %. Daunorubicin-resistance was found in 32/65 (49,2 %), resistance to both was observed in 6/65 (9,2 %). Treatment of daunorubicin-resistant lymphoblasts with idarubicin resulted in significant apoptosis induction in 26 out of 32 cell populations (81,3 %).

We clearly demonstrated here that the in vitro treatment of lymphoblasts from children with de novo or relapsed ALL with idarubicin induces significantly higher response rates than daunorubicin treatment. The ex vivo sensitivity of daunorubicin-resistant lymphoblasts of childhood ALL to idarubicin treatment reflects the better potency of idarubicin to induce apoptosis and to overcome daunorubicin resistance. These data prompted us to study the clinical relevance of idarubicin in ongoing clinical trials to improve existing therapeutic regiments. First clinical data point to a good tolerability of idarubicin in the treatment of relapsed ALL in childhood.
Assessment of the Frequency of Additional Malignancies in Patients with Splenic Marginal Zone Lymphoma (SMZL).
Authors: Iannitto Emilio , De Cantis Stefano , Minardi Viviana , Callea Vincenzo , Calvaruso Giuseppina , Mulè Antonino , Ammatuna Emanuele , Stelitano Caterina , Tripodo Claudio , Ambrosetti Achille , Pizzolo Giovanni , Florena Ada , Franco Vito , Florena Ada
Published: October 2019

Splenic marginal zone lymphoma (SMZL) is an indolent neoplasm, mostly occurring in the elderly, with a median survival time exceeding 10 years. The incidence of secondary cancer and late complications related to therapy have not been investigated. To assess the frequency of additional neoplasms in a series of consecutive SMZL patients and to estimate the corresponding risk of second cancers compared with that of the general population. we investigated the incidence of additional cancers in 129 patients diagnosed with SMZL in three italian hematological centers.

With a median follow-up time of 33.2 months, twelve additional invasive cancers were recorded (9.3%). The 3 and 5-year cumulative incidence rates of second malignancy were 5.7% and 19.4%, respectively. Five more patients (3.8%) had a diagnosis of malignant epithelial tumor prior to that of SMZL.

The incidence of second malignancies was slightly, but significantly, higher than the expected incidence rate. The Observed-to-Expected ratio (O/E) was 12/5.92=2.03, 95% confidence interval (CI) 1.05 to 3.56; p <0.05. Of the twelve second malignancies observed, four were urologic neoplasms ( O/E=4/1,08=3.70 (95% CI:1.01 to 9.48; p <0.05), four were lung cancers (O/E=4/0.44365=9.16; 95% CI: 1.41 to 13.25; p <0.05) and the other four diagnosis were of hepatic carcinoma (1), endometrial cancer (1), breast cancer (1), and colo-rectal cancer (1). Our findings evidence an increased frequency of additional neoplasms in patients with SMZL and suggest that this figure is significantly different than that expected in the general population. The incidence of cases with urinary tract and lung malignancies in our series is higher than expected. Although confirmatory data are needed, in our opinion SMZL patients are at risk of secondary maligancies and should be carefully investigated at diagnosis and monitored during the follow-up.
SDX-105 (Treanda™) Enhances the Tumor Growth Inhibitory Effect of Rituximab in Daudi Lymphoma Xenografts.
Authors: Kanekal Sarath , Crain Brian , Elliott Gary
Published: October 2019

SDX-105 (Treanda™, Bendamustine HCl), a structurally and mechanistically unique multifunctional anticancer drug, has been demonstrated to be more potent than other 2-chloroethylamine alkylators in a variety of cell lines resistant to traditional alkylating agents. We previously reported that SDX-105 inhibits growth of SUDHL-1 and Daudi human lymphoma tumor xenografts in SCID mice in a dose-dependent manner (AACR PROC 45: Abs #4575). The current studies examine whether SDX-105 in combination with rituximab confers better efficacy. Daudi cells (1.0 X 107 cells/mouse) were inoculated s.c. in the flanks of SCID mice. After the tumors reached approximately 100 mm3, SDX-105 (10 mg/kg/d i.p., d 0–4), rituximab (75 mg/kg/d i.v., d 0, 2, 4) or both (same respective doses and treatment days) were administered; a second cycle of treatment was subsequently initiated on day 28. Tumor growth and body weight were monitored for 60 days. On day 20, mean tumor volumes were 2118, 847, 511 and 221 mm3 for the control, rituximab, SDX-105 and the combination group, respectively. Tumors in treated groups were significantly smaller than tumors in the control group, and the tumors in the combination group were significantly smaller than tumors in the rituximab group (p <0.02). In the combination group, the antitumor effect was sustained up to day 60 and there was one ‘cure’ (non palpable). There was no body weight loss in the control or rituximab group; 10% body weight loss was observed in SDX-105 and the combination groups. These results corroborate our previous in vivo single agent data in this model and provide further support for the use of SDX-105 with rituximab in clinical trials.
Bronchial-Associated Lymphoid Tissue (BALT) Lymphoma: A Multicenter Retrospective Analysis.
Authors: Laszlo Daniele , Steffanoni Sara , Andreola Giovanna , Balzano Rosaria , Pruneri Giancarlo , Spaggiari Lorenzo , Crosta Cristiano , Zucca Emanuele , Martinelli Giovanni
Published: October 2019

Surgery has been the treatment of choice in patients affected by BALT NHL; however, radical surgery is not always possible and does not avoid the risk of relapse. Systemic treatment is recommended for symptomatic patients who relapse after surgery or cannot undergo such procedure; nevertheless, there is no standard systemic chemotherapy which could be recommended for such indolent disease. Here we report clinical data of 16 patients (9 men and 7 female) with biopsy-proven low grade BALT lymphoma followed at EIO and IOSI from 1992 to 2004. Median age was 62 years (range 50–77); 8 patients presented symptomatic disease at diagnosis. Thoracotomy was necessary to obtain histological material in eight patients, while in the other 8 no-invasive procedures were performed. Thirteen patients were considered in stage IE and 3 in stage IV (bone marrow involvement in 2 patients, gastric in 1), according to Ann Arbor. Five patients received upfront aggressive surgery: 1 underwent wedge resection and 4 lobectomy. Surgery was the definitive treatment for 3 patients. With a median follow-up of 17 months (range 9–31), all these patients are still alive with no evidence of disease. Twelve patients received systemic treatment: two because of non-radical surgery and ten because of advanced stage, symptomatic disease or psychosocial reasons. The majority of them (7 patients) received alkylating containing regimen (Chlorambucil ± PDN), alone or in combination with Rituximab (Mabthera) (2 patients). Three additional patients, refusing CT, received Rituximab alone in a standard schedule. Hematological and non-hematological toxicity was in general mild: transient G3 neutropenia occurred in only one patient. No patient discontinued treatment because of treatment-related toxicity and no toxic death was recorded. All patients responded to systemic therapy with five patients achieving CR. Four of them are still alive and free of disease after 24, 65, 27, 42 months respectively. One patients still in CR died for reasons different from lymphoma after 54 months. With a median follow-up of 60 months (range 31 – 63), only four patients developed progressive disease or relapse after 67, 5, 63, 18 months respectively (median TTP=40 months). Our experience suggests that systemic treatment, when appropriate, is able to achieve an high response rate in this relative uncommon disease. Considering the clinical results and the low degree of toxicity observed, alkylating containing regimen (Chlorambucil ± PDN) alone or in combination with monoclonal antibody could be considered as first line treatment for patients with BALT lymphoma.
Phase II Study of Alemtuzumab Treatment in Patients with Pretreated T-Cell Lymphoma.
Authors: Zinzani Pier Luigi , Tani Monica , Stefoni Vittorio , Alinari Lapo , Marchi Enrica , Gabriele Annalisa , Musuraca Gerardo , Fina Maria Paola , Pileri Stefano , Baccarani Michele
Published: October 2019

To evaluate the efficacy and toxcity of Alemtuzumab, an anti-CD52 monoclonal antibody in patients with relapsed or refractory cutaneous T-cell lymphomas and peripheral T-cell lymphomas. Between 2003 and March 2004, 10 previously treated patients with mycosis fungoides (MF; n=4) and peripheral T-cell lymphoma unspecified (PTCLU) with nodal involvement (n=6) were enrolled onto a phase II trial and treated with Alemtuzumab in our Institute. This monoclonal antibody was given at a dose of 10 mg 3 times a week for 4 consecutive weeks. Of the 10 patients, 2 (20%) achieved complete response (CR), 4 (40%) partial response (PR), and the remaining 4 showed no benefits from the treatment. In the MF subset were observed only PR (3/4, 75%), while in the PTCLU patients there were 2 (33%) CR and 1 (17%) PR, respectively. The durations of CRs were 3 and 8 months. Treatment was well tolerated; hematolgic toxicity was mild. In terms of infectious adverse events, cytomegalovirus reactivation occurred in 1 (10%) patient and none patient had additional suspect or manifest infection. The results of the present phase II study show activity of Alemtuzumab as a single agent in patients with pretreated CTCL and PTCL using a low dose schedule; at the same time, this efficacious reduced dose permitts to avoid treatment-related mortality and to curtail the CMV reactivation rate.
A New Multi-Parameter Flow Cytometric Assay for Monitoring Lymphoma Growth and Spread in a Pre-Clinical Murine Model for Human Lymphoma.
Authors: Neeson Paul J. , Paterson Yvonne
Published: October 2019

Murine lymphoma models are commonly used to test the efficacy of idiotype-based vaccines for B-cell lymphoma prior to human trials. Mouse survival has been used as an indirect measure of the tumor response to the vaccine. In this study, we describe a multiparameter flow cytometric (MPFC) assay that enables direct assessment of the tumor at the inoculation site and detection of lymphoma metastases. In initial experiments, we show that 38C13 mouse lymphoma cells expressed an aberrant phenotype compared to normal splenocytes. The combination of CD45, CD19, B220 and an anti-idiotypic antibody (S1C5) along with a sequential gating technique defined the 38C13 cells with high sensitivity and specificity. The specificity of the MPFC assay for 38C13 lymphoma cells was greater than a gating strategy using light scatter and idiotype antibody binding. The MPFC assay was used to monitor 38C13 tumor kinetics in the C3H/HeN mouse and demonstrated tumor growth at the inoculation site and metastases to the lymph nodes and bone marrow. The presence of 38C13 metastases in lymphoid organs and bone marrow predicted by the MPFC assay were confirmed by histology and immunohistochemistry (IHC). The MPFC assay will enable investigators to monitor 38C13 lymphoma tumor response to individual vaccines or other lymphoma therapy prior to clinical trials. Furthermore, the MPFC concept could be readily applied to other models of human B cell lymphoma.
Sigma 2-Agonist CB-64D Induces Apoptosis in Non Hodgkin’s Lymphoma Cells: Association with Decreased Akt Expression.
Authors: Dell’Agnola Chiara , Wojciechowski Wojciech , Marshall Shannon , Lopez Natalia E. , Li Huifen , Bowen Wayne D. , Espinoza-Delgado Igor J.
Published: October 2019

Background and aim: Sigma (σ) receptors are expressed at high density in a variety of tumor cell types, with an increased expression of σ2-subtype in rapidly proliferating tumors. CB-64D is a recently developed sigma agonist, showing a higher affinity for σ2- receptor subtype. We evaluated the cytotoxic activity of CB-64D in the extensively characterized Non-Hodgkin’s Lymphoma (NHL) cell lines DB, RL, and Ramos.

Materials and methods: Human DB, RL (both derived from patients with diffuse large cell lymphoma), and Ramos (Burkitt’s lymphoma) cells were exposed to increasing concentrations of CB-64D for various periods of time. Cells were harvested and stained with Annexin-V and PI and apoptosis was evaluated by flow cytometry. Expression of caspases, Akt and Bcl-2 was determined by Western blotting.

Results: CB-64D induced apoptosis in all NHL cells tested in both a time- and dose- dependent manner. Apoptosis was detected as early as 3 hours (7.5%) and reached its maximum at 12 hours (75%). As little as 5 μM CB-64D was able to induce apoptosis and maximum effects were noted with 40 μM CB-64D. To investigate the mechanisms responsible for the observed apoptosis, expression of caspases, Bcl-2, and Akt was evaluated. Western blot experiments revealed that CB-64D induced the expression of activated caspases 3, 8, and 9. Inhibitors of both caspase 3 and caspase 8 significantly decreased CB-64D-induced apoptosis. The induction of apoptosis was not associated with changes in Bcl-2 levels. In contrast, CB-64D induced a major decrease of Akt as determined by Western blot. Northern blot analysis demonstrated that Akt mRNA was not affected by CB-64D suggesting that the decreased expression of Akt was controlled at the post-transcriptional level. Treatment with the proteasome inhibitor PS-341 failed to prevent CB-64D-induced Akt down-modulation suggesting that the decreased expression of Akt is not due to proteasome degradation.

Conclusions: Our data demonstrate that the σ2-agonist CB-64D has a potent anti-lymphoma activity. CB-64D directly induces apoptosis through caspase-dependent mechanisms. Our data also show that CB-64D-induced apoptosis is associated with a decreased Akt expression that is controlled at the post-transcriptional level. The present results warrant the exploration of CB-64D in a pre-clinical model.
Investigation of FcγR Polymorphisms and Response to IL-2 (Proleukin®) and Rituximab Treatment in Rituximab-Resistant NHL Patients: Importance of the F/F Polymorphism at Position 158 of the Fcγ.
Authors: Milan Sandra , Wilson Susan E. , Kahn K.D. , Giordano Heidi
Published: October 2019

Background: Antibody-dependent cellular cytotoxicty (ADCC) is an important mechanism of rituximab activity. Genetic polymorphisms in FcγRIIIa have been reported to influence response to rituximab in follicular non-Hodgkin’s lymphoma (NHL) patients, and the F/F polymorphism at position 158 has been associated with decreased response (Cartron, et al. Blood 2004). IL-2 induces expansion and activation of FcR bearing cells, enhancing ADCC (Gluck, et al. Clin Cancer Res 2004).

Methods: The combination therapy of IL-2 and rituximab (IL-2+R) was evaluated in low grade NHL patients who were rituximab-resistant by standard criteria. Rituximab was administered weeks (wks) 1–4 at 375 mg/m2 IV; IL-2 was administered SC 3 times/wk at 14 MIU wks 2–5 and at 10 MIU wks 6–9.

Results: 57 patients (pts) were enrolled. Twenty-nine pts are evaluable with at least week 8 response data, all of whom have been genotyped. Best response to date: 4 responders (1 CR, 3 PR), 22 patients with stable disease (SD), and 3 pts have progressed (PD) on therapy. Follow-up is on-going. FcγRIIIa polymorphism analysis at position 158 from 36 pts typed to date shows a frequency of 53% F/F, 36% V/F, and 11% V/V as compared to 35%F/F, 45%V/F and 20% V/V in previously untreated pts (Cartron et al, 2004). To date, the data has shown a higher response rate for the F/F pts (4/17; 24%) than the overall population (4/29; 14%). Best response to date for 17 F/F pts: 4 responders and 13 SD; for the 9 V/F pts: 0 responders, 7 SD, and 2 PD; for the 3 V/V pts: 0 responders, 2 SD, and 1PD. In addition, 9 pts had responses lasting = 4 months with 6/9 being F/F and 3/9 being V/F. Patients will be followed periodically up to 2 years or until disease progression. The regimen was well tolerated and the majority of adverse events (AEs) were CTC grade 0, 1, or 2 (flu like symptoms, injection site reactions and rash). No grade 3 or 4 AEs occurred in 10% or more of pts.

Conclusions: These data support an association of the genetic polymorphism of FcγRIIIa 158F/F with prior failure to respond to rituximab. Immunotherapy with IL-2+R may achieve a critical threshold to drive ADCC more effectively in patients carrying the low affinity F/F IgG FcR polymorphism, restoring their response to monoclonal antibody therapy. All pts with this F/F polymorphism either responded to therapy or achieved SD as their best response to date. This outpatient regimen was generally well tolerated. The role of the FcγRIIIa F/F polymorphism is currently being evaluated in a prospective randomized trial comparing rituximab to rituximab + IL-2 in patients with follicular lymphoma.
Results of Multicentre Phase IV Study of Rituximab in Combination with CHOP Chemotherapy in Patients with Previously Untreated Non-Hodgkin’s Lymphoma.
Authors: Shen Ti , Guan Zhongzhen , Shen Zhixiang , She Yuankai , Zhu Jun
Published: October 2019

Purpose: Rituximab is a chimeric anti-CD20 monoclonal antibody that was the first antibody approved by the FDA in the United States of America and SDA in China for the treatment of B-cell non-Hodgkin’s Lymphoma (NHL). It has shown significant efficacy and good tolerability in refractory and relapsed NHL. We have conducted a multicenter phase IV trial to evaluate the efficacy and safety of rituximab combined with standard CHOP chemotherapy in patients with newly diagnosed B-NHL.

Methods: Patients with newly diagnosed, histologically proven CD20-positive NHL were eligible for the study. All patients received 4–6 infusions of rituximab (375mg/m2 per dose) in combination with CHOP chemotherapy, either concurrently (rituximab administered on the first day of each 21-day CHOP cycle) or sequentially (4–6 once-weekly infusions of rituximab followed by six 21-day cycles of CHOP). Each CHOP cycle consisted of cyclophosphamide 750 mg/m2, doxorubicin 50mg/m2, and vincristine 1.4mg/m2 (maximum 2.0mg/dose) given intravenously on day 1, and prednisone 100mg/day orally on days 1-5. Tumor responses were assessed at the end of treatment.

Results: A total of 347 patients were recruited between February 2002 and December 2003. Of these 235 (68%) were male and 94 (27%) aged >60. The main lymphoma subtypes were diffuse large B-cell 196 (56%), follicular 41(12%), small lymphocytic/chronic lymphocytic leukemia 13(4%) and MALT 11(3%). Ann Arbor staging was as follows: stage I, 52 (15%); stage II, 80 (23%); stage III, 90(26%); stage IV, 105(30%); twenty patients (6%) could not be assessed. Of the 347 patients enrolled, 314 were evaluable for response. An objective response was observed in 94% of evaluable patients with a complete response (CR) in 56%, stable disease in 3.8% and progressive disease in 2.5%. The complete response rate was 63% for patients receiving 6 cycles of rituximab and 54% for those receiving four cycles of rituximab. No difference in response rate was observed between the sequential and concurrent groups. The most common adverse events were leucopenia in 122 patients (35%), nausea and vomiting 66 (19%), fever 39 (11%), rash 15 (4%) and asthma 4 (1%).

Conclusion: The combination of rituximab and CHOP chemotherapy is an effective and well-tolerated treatment for patients with newly-diagnosed CD20-positive NHL. The safety and efficacy achieved in this study suggests that more than four doses of rituximab may be required for optimal efficacy.
Survival Benefits of Patients with Non-Hodgkin’s Lymphoma Treated with Rituximab Combined with Chemotherapy.
Authors: Shen Zhixiang , Li Junmin , Wang Aihua , Chen Yu
Published: October 2019

Purpose: Rituximab combined with chemotherapy has been recommended as first-line or second-line standard regimen in most subtypes of B-cell lymphoma in China by the 2004 National Comprehensive Cancer Network lymphoma therapy guideline. We have conducted a multicenter trial to evaluate the efficacy and safety of rituximab in combination with standard chemotherapy (CHOP) in patients with previously untreated or relapsed indolent and aggressive NHL.

Methods: Patients received 4–8 cycles of rituximab plus CHOP every 21 days. For each cycle, rituximab (375mg/m2) was given on day 1 and CHOP started on day 3. CHOP consisted of cyclophosphamide 750mg/m2, doxorubicin 50mg/m2, and vincristine 1.4mg/m2 (maximum 2mg/dose) given intravenously on day 3, and oral prednisone 100mg on days 3–7.

Results: A total of 221 patients were enrolled on the trial, 128 males and 93 females with a mean age of 49 years (range 10–83 years). The main lymphoma subtypes were small lymphocytic (15 patients, 7%), follicular (27 patients, 12%), and diffuse large B-cell (160 patients, 72%). In total, 56 patients had indolent NHL and 165 aggressive NHL. The overall response rate for all patients was 86% with 57% complete responses. In patients with indolent NHL the overall and complete response rates were 95% and 55% respectively. After a median 12 months follow up, progression-free survival in patients with indolent NHL was 88%±5% at 1 year and 83%±6% at 2 years. In the 160 patients with diffuse large B-cell lymphoma, the overall response rate was 88% with 61% complete responses, and after a mean follow-up of 6 months, predicted 1-year and 2-year progression-free survival were 88%±5% and 83%±7% respectively. Infusion-related adverse events occurred in 4% of patients, associated with the first infusion of rituximab. Subanalyses according to subtype, stage, IPI and other factors will be presented.

Conclusion: Rituximab plus chemotherapy is an effective, well-tolerated treatment that achieves high response rates and long progression-free survival in both indolent and aggressive NHL.
Analysis of Histone Deacetylase Inhibitor, FK228, Effect on Chronic Myelogenous Leukemia Cell Line.
Authors: Okabe Seiichi , Tauchi Testuzo , Nakajima Akihiro , Sashida Goro , Sumi Masahiki , Gotoh Akihiko , Ohyashiki Kazuma
Published: October 2019

Chronic myelogenous leukemia (CML) results from transformation of hematopoietic cells by the BCR/ABL gene. Although high rates of hematologic responses to imatinib therapy, the acquired resistance to imatinib has been recognized as a major problem in the treatment of CML Histone deacetylases (HDACs) and histone acetyltransferases (HATs) regulate gene expression and cell growth. Recently, HDAC inhibitors have known as a new class of anti-cancer drugs. One of the HDAC inhibitor, FK228 (FR901228, depsipeptide) is now doing the clinical trial for the treatment of patients, such as peripheral T-cell lymphoma, but there was not known to the CML. In this study, we used the TF-1 BCR-ABL cell line, which were transfected BCR/ABL gene to the leukemia cell line, TF-1. We show here that FK228 potently induced apoptosis of TF-1 BCR-ABL cells, compare to the parental cell line, TF-1, in a dose and time depend fashion. BCR-ABL, intracellular molecular chaperone, heat shock protein 90 (HSP90), and p53 which regulate cell cycle, were acetylated after FK228 treatment, but not glycogen synthase kinase-3 β(GSK-3β) and signal-transducing activators of transcription 5 (STAT5). Histone H4 is also acetylated after FK228 treatment. In a cell cycle analysis, TF-1 BCR-ABL cells were stopped at G2-M phase after FK228 treatment. The activity of MAPK and Src kinases were blocked after FK228 treatment in a time and dose depend fashion, but p38 was activated. Inhibitor of apoptosis proteins (c-IAPs) have prevented cell death by inhibiting effectors caspases. IAPs were inhibited by FK228 and caspase3, caspase9 and poly (ADP-ribose) polymerase (PARP) were activated in a time and dose depend manner. Histone acetylation and caspase activitation were not blocked by treatment of p38 inhibitor, SB203580. Our study supports the future clinical trial of FK228 in the management of CML patients.
Quantitative Polymerase Chain Reaction (qPCR) at Diagnosis and Follow-Up of Patients with Chronic Myeloid Leukemia in Treatment with Imatinib.
Authors: Vallespí Teresa , Borrego Mildred , Colomé Doulous , Jaen Ana , Rozman Maria , Massagué Isabel
Published: October 2019

Imatinib, a selective inhibitor of tyrosin-kinasa BCR/ABL fusion protein, produces high response rates in patients with chronic myeloid leukemia (CML) in the chronic phase. The behavior of the levels of the BCR/ABL transcript by qPCR technique was studied in 24 patients (13M/11F; median age: 49.6 years, range: 23–73), diagnosed of CML in chronic phase, before and after the treatment with imatinib (at the 0, 3, 7, 11 and 15 months). Eight patients had previously received interferon alpha or busulphan and 16 hidroxiurea. The amplification of BCR/ABL with the qPCR technique, according to protocol BIOMED-2, was carried out in samples of bone marrow (n = 23) and peripheral blood (n = 48). Results were calculated in relation to a control gene of the glucuronidasa (GUS) and expressed in logarithmic scale (log10). Median time from the diagnosis was of 28 months and median treatment time with imatinib was 14 months. The median reduction of BCR/ABL transcripts was 1,98 log after three months and decreased further (2,71 log) during follow-up. Both of them were significative when compare with base line levels (P <0.001). The reduction was greater in patients who received a dose ³ 400 mg/day (n=16) than those with 300 mg/day (n=8). Three patients reached a major molecular response (ratio BCR-ABL/GUS <0.005) up to 15 months of treatment. In conclusion: the determination of the BCR/ABL transcripts by qPCR contributes with valuable information about the effect of imatinib in patients with CML. The logarithmic reduction is higher in the beginning of the treatment and doses ³ 400 mg/day are associated with a greater reduction of the tumoral load.

Figure Figure
Use of Single Course IV Fludarabine in Treatment of Hypocellular Bone Marrow (Aplastic Anaemia and MDS) Patients.
Authors: Apte Shashikant J. , Vaidya Sujata U. , Shah Nikhil , Bhatwadekar Seema , Kalro Akash , Sayaji Bhavna , Suryavanshi Prashant , Kelkar Anjali J.
Published: October 2019

Young patients with Hypocellular Bone Marrow ideally need Allogenic Stem Cell Transplant or ATG + CYA + Steroids. In country like India these two treatment modalities are very expensive and very few can afford. We tried to study effect of single course of IV FLUDARABINE 30 mg/m2 x 5 days as an upfront immunosuppressive therapy for these patients.

Study period: January 2001 – April 2004.

Total patients – 29.

Age group: 17 – 47 years

Follow up: 4–37 months. Median follow up 15 months.

Haemogram range at diagnosis

Hb: 3–7.2 gm%, ANC: 100 – 600/cmm, Platelet: 3 – 11 x 10 9/L

Bone marrow aspirate & trephine biopsy showed Hypocellularity on all patients.

8/29 patients had dysplastic features on morphology.

3/29 patients had cytogenetic anamoly, two: del 7 & one: del 7q.

14/29 patients had normal cytogenetics.

12/29 patients no metaphases seen on Cytogenetics.

All cellular transfusions were 2500 rads irradiated.

11/29 ( 38 %) patients died in first six months, causes of mortality being infection and intracranial bleed.

10/29 ( 34 %) patients responded to treatment. They are transfusion independent ( Hb > 9 gm%), platelet > 50000/cmm and ANC > 1500/cmm. All three patients with cytogenetic anamoly responded.

8/29 ( 28 %) patients showed no response.

Larger data is needed to asses the efficiency of single course IV Fludarabine in patients with Hypocellular bone marrow. Single five day course of IV Fludarabine can be a cheaper immunosuppression, for these patient.

Cost: ALLO SCT - Rs. 10 lakh ( $ 20000), ATG + CYA + Steroid - Rs. 4 lakh ($ 3000)

IV Fludara 30 mg/m2 x 5 day – Rs. 40000/- ( $ 800)
Autoimmune Disease (AD) in Patients with Myelodysplastic Syndrome (MDS): A Retrospective Single Institution Study.
Authors: Ramanarayanan Jeyanthi , Baer Alan N. , Battiwalla Minoo , Ford Laurie A. , Wetzler Meir , Baer Maria R.
Published: October 2019

Autoimmune disease (AD) can manifest uncommonly either at the time of diagnosis of MDS or during its course. When present, AD generally responds to immunosuppressive therapies, but cytopenias and immunosuppression associated with MDS compromise delivery of these therapies. Few studies have investigated the impact of co-existing AD on the course and outcome of patients with MDS. Our objective was to evaluate the clinical manifestations, laboratory characteristics, response to therapy and survival of MDS patients with AD. Records of patients evaluated at Roswell Park Cancer Institute with pathologically demonstrated MDS between 1993 and 2003 (n=277) were reviewed and patients with evidence of AD were identified. Patients with laboratory abnormalities without disease manifestations were excluded, as were patients with therapy-related MDS following treatment for AD. 13 patients (4%) were identified with co-existing MDS and AD. The initial presentation was AD in 6 (46%) and MDS in 4 (31%), while 3 patients (23%) had near-simultaneous diagnoses of both conditions. The spectrum of AD in these patients included systemic vasculitis in 3 patients, systemic lupus erythematosus in two and rheumatoid arthritis, temporal arteritis, cryoglobulinemia, aphthous stomatitis, pyoderma gangrenosum, inflammatory bowel disease, erythema nodosum and Evans syndrome in one patient each. Anti-double stranded DNA (levels ≥ 40.0 u/ml; normal range 0.0–3.5u/ml), ANA (≥1:160), cold agglutinins, low C3 and elevated ESR (≥100mm/hr) were the serological abnormalities detected at the time of AD diagnosis. Eleven of 13 patients were female, and median age at diagnosis of MDS was 65 years, while the entire cohort was 44% female (p=0.005) and had a median age of 71 yrs at diagnosis. FAB subtypes were RA (n=7), RAEB (n=3), CMMoL (n=2) and RARS (n=1). Cytogenetics were normal in 5 patients; abnormalities in the other 8 patients included −7, +8, and del(5q). The median survival of patients from diagnosis of MDS was 48 months and the survival from diagnosis of AD was 46 months. Known causes of death in 6 patients included sepsis, intracranial hemorrhage, lung cancer and transplant-associated multiorgan failure. Based on this study, AD occurs in 4% of MDS patients, predominantly affects female patients, and has heterogeneous clinical manifestations.The pathobiologic implication of the occurrence of AD at the same time or after the diagnosis of MDS is that the dysplastic clone might be responsible for the induction of immune dysregulation.
Analysis of Transforming Growth Factor Receptor Type II in Patients with Agnogenic Myeloid Metaplasia (AMM).
Authors: Hemavathy Kirugaval C. , Novetsky Allan D. , Wang Jen C.
Published: October 2019

We previously reported that blood TIMP-1 (Tissue Inhibitor of Metalloproteinase), OPG (Osteoprotegerin), and TPO (Thrombopoietin) levels are increased and blood MMP-3, and bone marrow SDF-1 levels are decreased in patients with AMM. Enhanced TGF-ß 1 activity with Transforming Growth Factor Receptor Type II (TGF ß RII) abnormalities is likely to be associated with these findings. It has been previously reported that TGF-ß RII levels are decreased in AMM patients. The current study was undertaken to explore the defects in the TGF-ß1 pathway in this disease. We cloned and sequenced the full length TGF ß RII cDNA from AMM patients and no mutations were detected on sequencing. We then determined the expression levels of TGF ß RII in the CD34+ progenitor cells by Real-Time RT-PCR and found it to be reduced in patients with AMM compared to normal volunteer CD34+ cells used as controls. Hence we cloned the promoter region of TGF ß RII (−1233 to +50) from the genomic DNA of Peripheral Blood Monocytes by PCR, using gene specific primers. On sequencing, no mutations were detected in the promoter regions. The same promoter region when cloned upstream of a luciferase gene, drove the transcription of the reporter gene similar to the levels observed with the promoter from controls. This augmented the absence of any structural abnormalities in the region of the TGF b RII promoter. We then analyzed the methylation status of TGF ß RII promoter using the technique, Sodium Bisulfite-PCR for Sequencing. Genomic DNA was modified using Sodium Bisulfite and the TGF ß RII promoter was amplified by PCR with Primers capable of amplifying both unmethylated and methylated DNA. The modified PCR products were cloned into TOPO TA sequencing vector from Invitrogen, CA and sequenced. The sequencing results revealed absence of any CpG hyper-methylation in the promoter region of TGF ß RII. To account for reduced TGF ß RII expression in the absence of coding region abnormalities, promoter abnormalities or hypermethylation, we determined Histone Deacetylase Activity HDAC activity in the CD34+ cells of AMM patients. There was an indirect correlation between HDAC activity and TGF ß RII mRNA levels implying HDAC mediated silencing of the TGF ß RII promoter in AMM patients.
Plasma and Intracellular Concentration of TNF and Its Receptors in Peripheral Blood and Bone Marrow in Different Stages of B-CLL.
Authors: Rolinski Jacek , Bojarska-Junak Agnieszka , Hus Iwona , Dmoszynska Anna
Published: October 2019

TNF has been proposed to play a role in the regulation of growth and death of leukemic B-CLL cells. However, the biological effects of TNF on leukemic cells, as well as its role as a prognostic factor need to be further investigated. The aim of the study was to eevaluate the correlation of TNF and its receptors in peripheral blood (PB) and bone marrow (BM) with the stage of B-CLL and some other clinical parameters. PB and BM were taken from 44 newly diagnosed, untreated B-CLL. patients. The control group consisted of 20 healthy subjects. We used flow cytometry technique to assess the capability of T and B lymphocytes to produce TNF and ELISA method to measure plasma levels of TNF and their soluble receptors. We found, that PB and BM plasma TNF concentration in the patients was significantly higher than in the healthy control (2.61 pg/ml. vs 0.62 pg/ml; and 2.91 pg/ml vs 0.75 pg/ml, respectively p<0.001). TNF concentration in PB and BM was significantly higher in Rai stage III–IV than in early stages (p<0.01). There was a correlation between the PB and BM TNF level and lymphocytosis (p<0.005) and the total tumor mass (TTM) (p<0.0001). The PB and BM TNF concentration positively correlated with the percentage of T CD3+ lymphocytes producing intracellular TNF (p<0.01).

The percentage of T cells from PB an BM expressing cytoplasmic TNF was significantly higher in patients (PB:39.11±16.97%; BM:40.73±18.19%) than in normal controls (PB:15.74±7.95%; BM:18.80±12.93%) (p< 0.00001; p<0.005, respectively). In PB and BM from B-CLL patients the percentage of CD3+ cells expressing intracellular TNF was significantly higher than the percentage of CD19+/TNF+ cells (p<0.0001). Besides, it was found that the percentage of T cells expressing cytoplasmic TNF positively correlated with the stage of disease (p<0.01). In PB positive correlation were found between the number of T CD3+/TNF+ cells and lymphocytosis (p<0.05) and TTM (p<0.001). The percentage of leukaemic B cells positive for TNF did not correlate with the stage of disease.

There was increased expression of TNF-RI and TNF-RII in leukaemic B cells in comparison to normal B-cells was observed (p<0.0001). We found positive correlation between the number of CD5+ B lymphocytes and the levels of soluble TNF-RII (sTNF-RII) (p< 0.05). The sTNF-RII levels in PB and BM significantly correlated with the stage of disease acc. Rai (p<0.0001). Furthermore, the sTNF-RII concentration positively correlated with lymphocytosis and TTM (p<0.0001). These results strongly support the key role TNF in B-CLL pathogenesis. Our results suggest that TNF may function as growth factor for B-CLL cells. CD3+T cells may be the important source of this cytokine in advanced B-CLL. It seems that changes in T cells capability to produce cytoplasmic TNF are associated with disease progression. However, further studies are required to confirm the key role of TNF in B-CLL pathogenesis.
Fludarabine-Induced Apoptosis in Chronic Lymphocytic Leukemia Cells Is Not Affected by Inhibitor of Apoptosis Proteins (IAPS).
Authors: Silva Karina Lani , de Paula Castro Eric Delfraro , Vasconcellos Deborah Vidal , Kwee Jolie Kiemlian , Moellman Arthur Coelho , Klumb Claudete Esteves , Maia Raquel Ciuvalschi
Published: October 2019

B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the accumulation of mature lymphocytes due to defective apoptosis. Inhibition of apoptosis also contributes to chemoresistance, mainly in patients in advanced clinical stages. Fludarabine is a potent chemotherapeutic drug used for patients with B-CLL. Despite good initial clinical responses, some patients acquire resistance to this drug. One of the mechanisms through which malignant cells are believed to acquire resistance to apoptosis is by overexpression of inhibitor of apoptosis proteins (IAPs). IAPs are a family of proteins able to block apoptosis either by binding and inhibiting caspases or through caspases-independent mechanisms. To date, overexpression of IAPs has been detected in many types of malignant diseases, including B-CLL. As fludarabine can induce apoptosis trough caspases activation and this proteases could be inhibited by IAPs, we evaluated in this current study, the expression of three members of IAP family (c-IAP1, c-IAP2 and XIAP) by immunocitochemistry in 30 peripheral blood B-CLL specimens and correlated it to fludarabine-induced apoptosis. In parallel, we also evaluated the expression of these IAPs before and after in vitro fludarabine treatment. B-CLL cells were incubated with fludarabine25 m M for 12 hours and the induction of apoptosis were measured by annexin-V assay. The apoptosis index by fludarabine was extremely significant (p < 0.001). Different levels of c-IAP1, c-IAP2 and XIAP expression were commonly found in circulating B-CLL cells. High levels of c-IAP1, c-IAP2 and XIAP expression were observed in 20%, 60,7% and 60% of samples, respectively. However, the overexpression of these IAPs was not correlated with an in vitro fludarabine apoptosis-resistance. Besides, fludarabine was not able to change the levels of IAPs expression as described for other drugs in literature. These results suggest that fludarabine might cause apoptosis by a caspase-independent manner.
Gene Expression Profiling of Plasma Cell Dyscrasias: The Role of IGH Translocations in the Heterogeneity of Multiple Myeloma.
Authors: Neri Antonino , Mattioli Michela , Agnelli Luca , Fabris Sonia , Baldini Luca , Morabito Fortunato , Bicciato Silvio , Verdelli Donata , Intini Daniela , Nobili Lucia , Cro Lilla , Pruneri Giancarlo , Callea Vincenzo , Stelitano Caterina , Lombardi Luigia
Published: October 2019

Multiple Myeloma (MM) is the most common form of plasma cell dyscrasia, characterized by a marked heterogeneity of genetic lesions and clinical course. It may develop from a premalignant condition (monoclonal gammopathy ofundetermined significance, MGUS) or progress from intra-medullary to extra-medullaryforms (plasma cell leukemia, PCL). To provide insights into the molecular characterization of plasma cell dyscrasias and to investigate the contribution of specific genetic lesions to the biological and clinical heterogeneity of MM, we analyzed the gene expression profiles of plasma cells isolated from 7 MGUS, 39 MM and 6 PCL patients by means of DNA microarrays. MMs resulted highly heterogeneous at transcriptional level, whereas the differential expression of genes mainly involved in DNA metabolism and proliferation distinguished MGUS from PCLs and the majority of MM cases. The clustering of MM patients was mainly driven by the presence of the most recurrent translocations involving the immunoglobulin heavy-chain locus. Distinct signatures have been found to be associated with different lesions: the overexpression of CCND2 and genes involved in cell adhesion pathways was observed in cases with deregulated MAF and MAFB, whereas genes upregulated in cases with the t(4;14) showed apoptosis related functions. In addition, we identified a set of cancer germ-line antigens specifically expressed in a sub-group of MM patients characterized by an aggressive clinical evolution, a finding that could have implications for patient classification and immunotherapy.