bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of Two Regimens for the Treatment of Elderly Patients with Acute Lymphoblastic Leukaemia (ALL).
Authors: Offidani Massimo , Corvatta Laura , Malerba Lara , Marconi Monica , Catarini Massimo , Centurioni Riccardo , Leoni Franco , Scortechini Anna Rita , Masia Maria Cristiana , Leoni Pietro
Published: April 2020

Acute lymphoblastic leukemia (ALL) represents a rare malignancy in the elderly and few authors have specifically focused on the treatment of ALL in this setting. We recently published the results of a prospective phase II study comprising an induction therapy with vincristine, Daunoxome, cyclophosphamide and prednisone (VDXD) given to 15 patients aged ≥ 60 years. Here, we updated the results after enrolling 17 patients treated from 1999 to 2002 (median age 69 years, range 61–79) and we compared these with the results obtained in 17 elderly patients treated according to the GIMEMA ALL 0288 protocol from 1994 to 1998 (median age 70, range 61–76). Most of patients in both groups had B-lineage ALL and an initial WBC count less than 30 x 109/L. The most frequent karyotypic abnormality resulted the Ph which was detected in 4 (23%) and in 3 patients (17%) treated with 0288 and VDXD, respectively. BCR-ABL translocation was present in 45 % of patients treated with VDXD. Comorbidities were documented in 11 (65 %) patients treated with VDXD compared with 9 (58%) receiving 0288 regimen.

With VDXD combination elderly ALL had a higher CR rate than with 0288 protocol (76.5% vs 41%; p=0.037) likely due to both lower induction mortality (17.5% vs 35%; p=0.028) and less resistant disease (6% vs 24%; p=0.025). Infectious complications rate was similar (64% vs 53%) but they were the primary cause of death in 2 and 5 patients receiving VDXD and 0288, respectively. Non-hematological side effects were comparable. Median DFS was similar and resulted 18 months and 20 months with 0288 and VDXD treatment, respectively. Median EFS was 3.9 and 12.8 months, respectively (p = 0.0486). Particularly, 1-year EFS resulted 35% in patients receiving 0288 treatment compared with 53% in those given VDXD. Median OS was 4.5 an 21 months in the first and in the latter group of patients (p = 0.0239) with a two-year OS of 29% in 0288 and 38% in VDXD group.

Our results are encouraging and show that the administration of high-dose daunorubicin as liposomal compound in elderly ALL patients is not only feasible but also able to improve CR rate, EFS and OS without increase in toxicity.
Intensification of Post-Remissional Treatment in Adult Acute Lymphoblastic Leukemia (ALL) with t(4;11) or t(1;19): Results of the GIMEMA LAL 2000 Study.
Authors: Vignetti Marco , Meloni Giovanna , Morselli Monica , Tedeschi Alessandra , Ferrara Felicetto , Cantore Nicola , Specchia Giorgina , Di Raimondo Francesco , Recchia Anna , Sica Simona , Cascavilla Nicola , Camera Andrea , Fabbiano Francesco , Cimino Giuseppe , Mancini Marco , Fazi Paola , Foà Roberto , Mandelli Franco
Published: April 2020

In the GIMEMA LAL 0496 trial, which has been conducted in Italy between 1996 and 2000, patients with adult ALL received an induction therapy with VCR, DNR and ASP until they reached complete remission (CR). Consolidation therapy with Ara-C and CTX was then administered and, if still in CR, patients received a 3-year maintenance therapy. Ph+ ALL patients were considered at high risk and after the induction phase a stem cell transplant (SCT) was planned, allogeneic for patients with an HLA identical sibling or autologous. A total of 498 patients have been enrolled by 50 GIMEMA Centers. Of these, 29 patients (5.8%) with either t(4;11) or t(1;19), who received the standard post-remissional approach, showed a statistically significant worse prognosis after CR: the projected probabilities of DFS and OS at 2 years were 19% (C.I. 95%: 4.1–34.4) and 31% (C.I. 95%: 14.2–47.9), respectively. Thus, in the new protocol - GIMEMA LAL 2000 - these patients received an intensive post-remissional treatment including SCT. Between 2000 and 2003, a total of 383 patients have been enrolled in this new protocol. Of these, 23 (6.0%) were positive either for t(4;11) or for t(1;19). The clinical characteristics of these patients are comparable to those of the patients enrolled in the previous study and the overall CR rate is also super imposable.

GIMEMA LAL 0496 vs GIMEMA LAL 2000

Fig. 1 shows the comparison in DFS at 2 years between LAL 0496 and LAL 2000; though not statistically significant, a trend towards a better prognosis is clearly seen. The DFS probability projected at 2 years is 52% (95% C.I. 26–78) compared to 19% (95% C.I. 4–35). The OS, projected at 2 years, is 42% (95% C.I. 14–70) compared to 31% (C.I. 95%: 14.2–47.9).

Despite the relatively limited case series, these results suggest that for t(4;11)+ and t(1;19)+ adult ALL intensification of the post-remissional treatment with SCT is a feasible approach that may improve the outcome in this subset of patients at high risk of relapse with standard post-remissional therapies.

Figure Figure GIMEMA LAL 0496 GIMEMA LAL 2000 p Patients t(4;11) or t(1;19) 29 23 0.7626 Sex (M/F) 14/15 8/15 0.3280 Age yrs, median (min-max) 37.8 (14.9–59.4) 39.4 (19.6–59.2) 0.1481 WBC x 109/L median (min-max) 41.2 (2.2–663.0) 50.3 (2.4–300.0) 0.9193 Phenotype (B/T) 29/0 23/0 - CR rate 92.8% 85.7% 0.6392
Pioglitazone Inhibits the Growth of Human Leukemic Cell Lines and Primary Leukemic Cells in Vitro.
Authors: Hatta Yoshihiro , Saiki Minoru , Enomoto Yuko , Aizawa Shin , Sawada Umihiko , Horie Takashi
Published: April 2020

Troglitazone and pioglitazone are one of thiazolidinediones that are high affinity ligand for the nuclear receptor called peroxisome proliferator-activated receptor gamma (PPAR-γ). Troglitazone is a potent inhibitor of clonogenic growth of acute myeloid leukemia cells when combined with a retinoid. However, the effect of pioglitazone to neoplastic cells and normal hematopoietic cells has not been studied yet. Adult T-cell leukemia (ATL), prevalent in western Japan, is a highly aggressive malignancy of mature T lymphocyte. Therefore, we studied antitumor effect of pioglitazone against leukemic cells including ATL as well as normal hematopoietic cells. With 300 μM of pioglitazone, colony formation of ATL cell lines (MT1, MT2, F6T, OKM3T, and Su9T01) was completely inhibited. Colony formation of HUT102, another ATL cell line, was 12 % compared to untreated control. Clonogenic cells of other leukemic cell lines (K562, HL60, U937, HEL, CEM, and NALM1) was also inhibited to 0–30% of control. Colony formation of primary leukemic cells from 5 AML patients was decreased to 15 %. However, normal hematopoietic cells were weakly inhibited with 300 μM pioglitazone; 77 % of CFU-GM, 70 % of CFU-E, and 33 % of BFU-E survived. Cell cycle analysis showed that pioglitazone decreased the ratio of G2/M phase in HL60 cells, suggesting the inhibition of cell division. By Western blotting, PPAR-γ protein level was similar in all leukemic cells and normal bone marrow mononuclear cells. Taken together, pioglitazone effectively eliminate leukemic cells and could be used as an antitumor agent in vivo.
Anticancer Activity and Induction of Multinucleation in Hematologic Cell Lines Mediated by Ganoderma Lucidum.
Authors: Muller Claudia I. , Kumagai Takashi , O’Kelly James , Seeram Navindra , Heber David , Koeffler H. Phillip
Published: April 2020

Over many centuries, herbal remedies have treated a variety of ailments. This empiric observational approach has produced a number of leads for formulated medicines. We have examined five phytochemical herbs with purported anti-cancer activities: Ganoderma lucidum, Isatis indigotica, Dendranthema morifolium, Rabdosia rubescens and Panax pseudoginseng. Ethanol and DMSO extracts were made of each herb, and these were screened for their antiproliferative activities (MTT-assay; doses: 50 μg/ml and 100 μg/ml) using a panel of 26 human cancer cell lines (16 hematologic cell lines). Ganoderma lucidum was the most effective agent, particularly against the 16 hematological cell lines. Further dose- and time-response MTT assays of Ganoderma were performed using the 7 most sensitive hematological cell lines: their ED50’s were 26 μg/ml, HL-60; 63 μg/ml, U937; 50 μg/ml, K562 (all three are myeloid leukemias); 38 μg/ml, Blin-1; 30 μg/ml, Nalm-6 (both are B-ALLs); 40 μg/ml, RPMI8226 (multiple myeloma); 20 μg/ml, Daudi (Burkitt’s lymphoma). Cell cycle analysis revealed a G2/M arrest, most prominently in HL-60 cells (24% in G2/M compared to 12% in control cells cultured with 100 mg/ml Ganoderma for 3 days). Three hematological cell lines (HL-60, U937, Blin-1) were examined for apoptosis, which ranged between 10% – 35% and 30% – 49% compared to <6% in control cells after exposure to 100 and 200 mg/ml Ganoderma for 3 days, respectively (Annexin V staining). Of particular note, cytospin preparations of HL-60 cells revealed that Ganoderma lucidum caused prominent multinucleation. Dose-dependent studies revealed 1% multinucleation in the cells treated with 1 mg/ml, 5% with 10 mg/ml, 10% with 50 mg/ml, 40% with 75 mg/ml and 80% with 100 mg/ml Ganoderma at day 4 of culture. FACS analysis of PI stained HL-60 cells confirmed the increased DNA content of these cells. Of interest, multinucleation did not occur in 6 other hematologic and 6 solid tumor cell lines. In summary, we discovered that 5 phytochemical herbs have growth inhibitory effects on AML, CML, B-ALL, Burkitt’s lymphoma and multiple myeloma cell lines. The most effective drug was Ganoderma lucidum, and it induced cell death by apoptosis. Moreover, multinucleation was observed in HL-60 cells, which may provide interesting insight in the same process in normal cells, like megakaryocytes. Notably, these studies have identified one plant, Ganoderma lucidum, that deserves careful separation techniques to identify its anticancer molecule (s).
ATRA Based (Iraqi) Protocol for Children with Acute Promyelocytic Leukemia (APL) in Baghdad: Results in the First 8 Patients.
Authors: Testi Anna Maria , Moleti Maria Luisa , Nal-Hadad Salma Abbas , Al-Jadiry Mazin Faisal Farhan , Mandelli Franco , Foa Robert
Published: April 2020

At the Oncology Unit/Al Mansour Teaching Hospital for Pediatrics in Baghdad, Iraq, between January 2002 and January 2003, a total of 11 children (age < 15 years) out of 32 acute myelogenous leukaemia (AML), were morphologically diagnosed as APL. This represents an overall very high incidence (35%) of childhood AML referred to a single institution. Molecular and cytogenetic studies were not feasible. ATRA was not available and most children received combination cytarabine and daunorubicin induction treatment that resulted in more than 50% early fatalities mainly due to hemorrhagic events and in less than 10% survivors. As a consequence of a collaborative project between the Oncology Unit/Al Mansour Teaching Hospital for Pediatrics in Baghdad and the Pediatric Hematology Unit of our Institute at “La Sapienza” University of Rome, a specific ATRA based protocol was designed according to the modern strategies for the management of APL and adapted to the severe local difficulties in Iraq. ATRA was supplied by Roche. The treatment plan consisted of oral induction ATRA (25 mg/m2/day) for 30 days associated to Daunorubicin (DNR 25 mg/m2/day for 2 consecutive days) only if the WBC count was > 10 x 109/L at diagnosis or increased during treatment. Consolidation included three chemotherapy cycles (DNR; DNR+ARA-C; DNR); ATRA was associated to each cycle for patients with a WBC count at diagnosis > 10 x 109/L and/or for those requiring the addition of DNR during ATRA therapy (high-risk). Standard 6-MP and MTX maintenance with 14 days ATRA, every 3 months was administered to all patients in complete remission (CR) for 2 years. Intrathecal MTX prophylaxis (total 3 doses) was given at each consolidation course, for high-risk patients.

Since December 2003, 8 consecutive children with APL (5 male and 3 female; median age: 12 years, range 3–16) were enrolled in the protocol. Seven were defined as high-risk and one as low-risk. Morphological diagnosis was that of classic M3 in 7 children and of M3 variant in 1. The M3v child had a very high WBC count and died the same day of referral due to a massive bleeding, while the other 7 children achieved a CR. One of the 7 responders was lost to follow-up in CR; the other 6 patients are alive and well in CR, on consolidation (1 case) and on maintenance treatment (5 cases).

These preliminary but highly encouraging results represent a substantial improvement over the earlier experience in childhood APL in the main Pediatric Center in Baghdad and demonstrate that modern therapeutic strategies, adapted to the local reality, are a necessity and can be effectively implemented through international collaborative efforts even in countries with limited resources and severe difficulties.
Intracranial Hemorrhage (ICH) in Patients (pts) with Newly Diagnosed Leukemia: Incidence and Effect on Outcome.
Authors: Oki Yasuhiro , Jabbour Elias , Kantarjian Hagop , Estey Elihu , Cortes Jorge E. , Faderl Stefan , Pierce Sherry , Zhou Xian , Bekele B. Nebiyou , Garcia-Manero Guillermo
Published: April 2020

Hemorrhage and infection carry significant morbidity and mortality in pts with leukemia. ICH can be a major clinical problem despite the current advances in supportive care. To analyze the incidence, outcome of ICH, and to identify potential risk indicators, we have conducted a retrospective analysis of pts with leukemia at M.D. Anderson Cancer Center (MDACC). We first reviewed all brain CTs of pts newly referred to MDACC from 1/2003 to 11/2003 with diagnosis of leukemia, myelodysplastic syndrome, or myeloproliferative disorder. Among 1185 pts (275 AML, 78 ALL, 163 CML, 296 CLL, 206 MDS, 47 MPD, 111 Other), 13 (1%) had ICH (8 AML, 3 APL, and 2 CML-myeloid blastic crisis). Based on this preliminary analysis, we limited this analysis to pts with newly diagnosed AML who received induction chemotherapy between 1/1998 and 11/2003. Among 993 such pts, 28 (3%) had ICH (parenchymal bleed in 12, subarachnoid hemorrhage in 4, subdural hematoma in 6, hemorrhagic infarct in 4, subarachnoid hemorrhage + subdural hematoma in 1, subarachnoid hemorrhage + subdural hemorrhage + parenchymal hemorrhage in 1) during the induction period (within 6 months after induction chemotherapy). Pt characteristics are shown in the table. Median time from induction therapy to ICH was 27 (0–122) days. In 16 (57%) pts death occurred before response to therapy could be determined (0–37 days). Thirteen out of 16 (81%) pts died of ICH as an immediate cause of death, and three of 16 (19%) died in the setting of multi-organ failure. Of 12 remaining pts whose disease status was assessed, 8 pts (66%) achieved complete remission (CR). Univariate analysis including age, gender, APL vs non-APL, cytogenetics, performance status, protective environment treatment, presence of antecedent hematologic disorder (AHD) and/or MDS, WBC< hemoglobin, platelet, PT, PTT, b2MG, bilirubin, creatinine, LDH, % of bone marrow blasts, % of peripheral blood blasts, and CR vs non-CR, indicated that APL, cytogenetics, thrombocytopenia, PT, bilirubin, and non-CR were associated with ICH. Differences in PT and bilirubin between the two groups, however, were clinically not significant. In a multivariate logistic regression analysis, APL remained as the only major predictive factor (odds ratio 3.35). In conclusion, ICH is limited to pts with myeloid malignancies, with an incidence of 3% in AML pts receiving induction therapy, with APL pts (13%) at greatest risk. ICH is associated with poor prognosis related to immediate mortality.

Characteristics Age Male APL Platelet (x109/L) CR rate Median Survival ICH (n=28) 56 (36–84) 50% 29% 33(4–166) 29% 5 weeks No-ICH (n=965) 58 (17–89) 41% 6% 47(4–635) 54% 42 weeks
Resistance to the Farnesyl Transferase Inhibitor SCH66336/Lonafarnib Caused by Mutations in the Target Protein Farnesyl Transferase Beta.
Authors: Raz Tal , Mohammad Azam , Daley George Q.
Published: April 2020

Farnesyl transferase inhibitors (FTIs) are target specific anti-cancer agents in clinical trials for the treatment of multiple myeloma, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), breast cancer, and other malignancies. FTIs were developed to block Ras prenylation; however, their cytostatic properties likely relate to activity against multiple farnesylated targets. The FTI Lonafarnib is showing activity against CML both as a single agent and in combination with the ABL-kinase inhibitor Gleevec. Though Gleevec is remarkably effective, treatment selects for drug-resistant mutations in the BCR/ABL target oncoprotein. Previously, we performed in vitro mutagenesis of BCR/ABL and identified a large catalogue of resistance mutations, anticipating essentially all mutations that have proven to be clinically problematic (Azam et al., Cell 2002). Thus we set out to determine whether FTIs would select for resistance mutations in Farnesyl Transferase Beta (FTase β), a normal target protein in the cell. We mutagenized (FTase β) by plasmid propagation in mutagenic E. coli, thus generating a random, unbiased library of mutated FTase β. This library was then introduced into K-Ras expressing BaF3 cells, which rely on K-Ras activity for cell survival, and undergo apoptotic cell death when treated with Lonafarnib. K-Ras transformed cells carrying mutated FTase (KR/MutFTB) were plated in soft agar in varying Lonafarnib concentrations. Lonafarnib-resistant colonies were harvested, and plasmids carrying mutant FTase βwere recovered and sequenced. Thus far we have identified 14 mutations that confer resistance to Lonafarnib. Among the mutations is W361L, previously shown to confer resistance to SCH56582 and SCH44342, compounds which are structurally similar to Lonafarnib (Del Villar et al. 1999). Most but not all mutations are positioned in the active site of the enzyme. To verify the drug resistant phenotype, 7 mutations were recreated de-novo by site-directed mutagenesis and transduced into BaF3-KRas cells. These cells were tested by a colony forming assay in the presence of drug. FTase activity in the presence of drug was also assessed by immunoblotting of HDJ2, a farnesylated protein. Cells transduced by mutated FTase showed increased ability to grow in the presence of Lonafarnib compared to controls. Interestingly, protein analysis showed reduced inhibition of farnesylation of HDJ2 only in the 361 mutants. This suggests that the mutant FTase may prenylate only a select set of substrates (not including HDJ2), a possibility consistent with previous reports of altered substrate specificity in FTB mutants in yeast (Del Villar et al. 1997). This catalogue of Lonafarnib-resistant FTase mutations will be useful for determining mechanisms of disease refractoriness or relapse in clinical trials, and can be used to profile drug activity of other farnesyl transferase inhibitors to find non-cross-resistant drug combinations.
Reversion of Chemoresistance in Aggressive Lymphoma by Quinine. A Phase II Trial.
Authors: Soubeyran Pierre , Masmoudi Amine , Blanc-Bisson Christele , Soubeyran Isabelle , Bellott Ricardo , Monnereau Alain , Eghbali Houchingue , Robert Jacques
Published: April 2020

Multidrug resistance has been one of the most studied mechanisms of cancer cell resistance to chemotherapy. Evidence of its involvement in hematological malignancies is known since many years. One well-designed phase I trial showed that about 20 to 25% of patients strictly resistant to EPOCH schedule could become responsive again by the addition of Dex-verapamil (J Clin Oncol1995; 13:1995–2004). We decided to perform a phase II trial to evaluate whether quinine could reverse resistance to chemotherapy in aggressive lymphoma. Methods: Patients included had all received at least two lines of chemotherapy including CEOP (cyclophosphamide 750mg/m2, epirubicine 60mg/m2, vincristine 1.4mg/m2 IV day 1 and prednisone 40mg/m2 orally day1 to day 7) and were demonstrated as strictly refractory to CEOP i.e. either no change or progression. Q-CEOP was administered as follows: quinine 30-mg/kg/day for two days from d0 to d1 as a continuous infusion, CEOP at the beginning of d1. Results: 15 patients have been included with the following characteristics: median age: 52 years (32–64), 66% stage III–IV, 60% elevated LDH. The median number of previous regimens was 3 (2–5). 13 patients had diffuse large B cell lymphoma including 3 with previous history of follicular lymphoma. Two had mantle cell lymphoma. Seven patients were included directly since they were initially refractory to CEOP while eight were shown refractory after two further cycles of CEOP. Epirubicin pharmacokinetics was not influenced by quinine. Mean dose intensity of epirubicin was 94%. Toxicity was mild. 4 responses were observed at the end of treatment including 2 complete and 2 partial responses (26%, IC 95%: 8–55%). Only the 2 CR patients are still alive with no evidence of disease at 35+ and 61+ months. Conclusion: These results add further arguments to confirm that MDR1 plays a significant role in resistance to chemotherapy in lymphoma. Quinine allowed unexpected long term remission in 2 responding patients.
A Pilot Study of Combined Escalated BEACOPP-ABVD Therapy for Advanced Hodgkin’s Lymphoma Patients with High IPS Score: The Israel Cooperative Lymphoma Group.
Authors: Avigdor Abraham , Bulvik Shlomo , Shemtov Noga , Levi Itai , Berkowicz Miriam , Ben-Yehuda Dina , Blickstein Dorit , Goldschmidt Neta , Gural Alexander , Kornberg Abraham , Libster Dianna , Merkel Drorit , Naparstek Elizabeth , Raanani Pia , Roth Batia , Volchek Yulia , Nagler Arnon , Ben-Bassat Isaac , Polliack Aaron
Published: April 2020

ABVD is widely considered as the gold standard treatment for advanced Hodgkin’s lymphoma (HL), although about 40% of patients relapse or do not respond to initial treatment. Recently, the HD9 trial of the German Hodgkin’s Lymphoma Study Group has shown that escalated (esc) BEACOPP regimen can achieve better disease control than ABVD, but has a high incidence of acute and long-term toxicities including high occurrence of AML/MDS. In an attempt to decrease toxicity while preserving the potential benefit of upfront intensive therapy, we conducted a pilot study, which tested the feasibility, toxicity and efficacy of combined escBEACOPP-ABVD regimen as therapy for newly diagnosed patients with high risk (IPS≥3) stage III–IV HL. Patients initially received 2 cycles of escBEACOPP followed by reevaluation with CT and gallium or PET/CT-FDG scans. When complete response (CR) or partial response (PR) was achieved, patients continued to receive 4 cycles of ABVD, while those failing to achieve a response were withdrawn from the study. Since August 2001, 21 patients entered the study and at the time of present analysis four of them are still receiving therapy. Three (18%) of 17 patients, who completed chemotherapy, received consolidative radiotherapy. Median age at diagnosis was 26 years (range 18–56) and 11 (57%) were males. Stage IV and B symptoms were evidenced in 19 (90%) and 17 (81%), respectively, and 7 (33%) had bulky mediastinal mass. Histology included nodular sclerosis in 18 patients (86%), mixed cellularity in 2 (10%) and lymphocyte predominance in 1 (5%). Following the first 2 cycles of escBEACOPP the overall response rate (CR+PR) was 100%. At the end of all therapy 15 patients (88%) were in CR, one patient in PR and only a single patient had progressive disease. With a median follow-up of 20 months no patient relapsed or died. Toxicity included WHO grade III–IV granulocytopenia in 15 patients (88%) and grade III–IV infection in one patient. Hospitalization for intravenous antibiotics was necessary in 10 patients (59%). Almost all of these events occurred during the first two cycles of escBEACOPP, while acute toxicity during the ABVD phase was mild. In conclusion, the combined escBEACOPP-ABVD regimen is well tolerated and is associated with a high CR rate when used in advanced HL patients with high IPS scores. Further follow-up is obviously required in order to determine long-term survival and late complications of this regimen.
A Phase II Trial of Dexamethasone, High-Dose Cytarabine, and Cisplatin (DHAP) in Combination with Rituximab as Salvage Treatment for Patients with Refractory or Relapsed Aggressive Non-Hodgkin’s Lymphoma.
Authors: Mey Ulrich J.M. , Strehl John W. , Orlopp Katjana S. , Flieger Dimitri , Birkmann J. , Ho Anthony , Kaiser U. , Neubauer Andreas , Glasmacher Axel G. , Schmidt-Wolf Ingo G.H.
Published: April 2020

Purpose: With commonly used salvage chemotherapy regimens like DHAP, response and overall survival rates are unsatisfactory. We designed a multicenter phase II trial to evaluate the safety and efficacy of the combination of rituximab with the DHAP regimen in patients who relapsed after or were resistant to a CHOP-like regimen.

Methods: 61 patients with relapsed or resistant aggressive B-cell NHL were included in this trial so far. The median age was 61,5 years. 39 patients were in first relapse; 21 had recurrence during the first 12 months after treatment. 5 patients had second or subsequent relapse, 17 had primary refractory disease. Treatment consisted of rituximab infusions (375 mg/m2 per dose) on day 1 followed by dexamethasone 40 mg d3–6 (d 3–5 in first cycle), cytarabine 2 x 2000 mg/m2 d 4 (2 x 1000 mg/m2 in first cycle), 2 x 1000 mg/m2 for patients > 60 years of age (2 x 500 mg/m2 in first cycle), and cisplatin 25 mg/m2 d 3–6 (d3–5 in first cycle) for a maximum of 4 cycles. When hematological toxicity did not allow continuation of chemotherapy on day 22, rituximab was given as single agent on day 22 and the next cycle was postponed until day 29.

Results: The overall response rate in evaluable patients was 54.0%. 29.0% of patients achieved a complete response, 25% of patients had a partial response, 15.0% of patients had stable disease. 31.0% of patients showed progressive disease. Patients in first relapse achieved an overall response of 67.0 % as opposed to only 27.0% in patients with refractory disease. Grade 3/ 4 nausea and vomiting were the only severe toxicities attributed to rituximab in one patient. Main toxicities were neutropenia and thrombocytopenia. Grade 3/ 4 nephrotoxicity occured in 2 patients. Survival data will be presented at the meeting.

Conclusions: Interim analysis of the data suggests that the combination of rituximab with the DHAP regimen in the treatment of relapsed or refractory aggressive lymphoma is feasible and effective. In view of the unfavorable prognostic factors in our patient population, response rates (ORR = 54%) are encouraging and warrant further investigation.
Epstein-Barr Viral Load in Post Renal Transplant Lymphoproliferative Disease: Elevated EBV DNA Levels at Diagnosis and Clearence after Rituximab Therapy.
Authors: Baiocchi Otávio C.G. , Caballero Otávia L. , Vettore André L. , Bulgarelli Adriana , Franco Marcello F. , Alves Antonio C. , Pestana José O.M. , Colleoni Gisele W.B. , Bordin José O.
Published: April 2020

The imbalance between EBV load and host immunological status is the trigger for post transplant lymphoproliferative disease (PTLD) development. Thus, monitorization of EBV load can be a useful tool in the management of these patients. In order to evaluate this issue in a renal transplant center, we studied eleven PTLD cases between January 2001 and December 2003. The diagnosis of PTLD was established according to WHO classification and confirmed by in situ hybridization (ISH) for EBV. Real-time PCR (LightCycler System, Roche, USA) and primers for amplification of EBV’s EBNA-1 gene were used to evaluate EBV viral load in peripheral mononuclear cells (PBMC) of PTLD cases. Samples were collected at diagnosis and 1, 3, 6 and 9 months after therapy. Median time from renal transplant to PTLD onset was 36 months. Eight (73%) were classified as diffuse large B-cell lymphoma, one as lymphoplasmocytoid lymphoma, one as polymorphic hiperplasia and one as Hodgkin’s lymphoma. All PTLD cases presented positive IgG and negative IgM antibodies, indicating remote EBV infection. All patients had their immunosuppression reduced (n=11), 5 received chemotherapy and 6 received monotherapy with Rituximab. Six patients (54%) achieved complete remission (CR), interestingly those with IPI <=2, with a median follow-up of 20.5 months. Six (54%) patients died (four of PTLD and two of sepsis). At diagnosis, median EBV viral load of PTLD cases was 17,400 copies/106 PBMC (range 1,000 to 227,590). All six patients who achieved CR cleared their viral load one month after therapy initiation, independent of EBV levels at diagnosis and kept it undetectable during follow-up. Five out of six patients who received Rituximab achieved CR. We conclude that clearance of EBV load occurs early after therapy start and that its maintenance probably depends on restoration of T-cell defense after immunosuppression reduction. We also concluded that Rituximab monotherapy may be a reasonable therapy with a low toxicity profile in this group of patients.
Proteasome Inhibitors Can Induce Caspase-3 Activation and Apoptosis in Human CML Cell Lines.
Authors: Kim Byung-Su , Kim Chang Up , Kim Young-Ju , Bae Eun Kyung , Kim Jinhee , Ahn Kwang-Sung , Park Seonyang , Kim Byoung Kook , Yoon Sung-Soo
Published: April 2020

The proteasome is a multi-enzyme complex that provides the ubiquitin-dependent degradation of many cytoplasmic and nuclear proteins involved in cell cycle progression and apoptosis. Inhibition of the proteasome represents a promising approach for the treatment of cancer because it can lead to cell cycle arrest and activation of caspases in tumor cells. There are several proteasome inhibitors that have been reported to induce apoptosis in various tumors. However, the effect of proteasome inhibition in human myeloid leukemia has not been reported so far. In this study, we tested two peptide-aldehyde proteasome inhibitors (MG115, MG132) on two human CML cell lines (K562, KCL22). At first, we treated both cell lines for 24, 48 and 72 hours with different doses of MG115 and MG132 and cell viability was tested by MTT assay. It showed substantial time and dose dependent cytotoxicity in both CML cell lines. Acridine orange staining also revealed DNA fragmentation. We then performed caspase-3 colorimetric assay after treating both cell lines for 6, 12 and 24 hours with 0.78μM of MG115, MG132. K562 showed the continuous rising of caspase-3 activity, while KCL22 exhibited the initial increase and subsequent mild decrease of caspase-3 activity. In addition, western blot analysis showed the reduction of procaspase-3 expression. The expression of Bcl-2 and Bcl-XL was reduced by western blot. p21 expression was slightly increased and that of cyclin D1 was decreased. Additionally, the treatment of proteasome inhibitor in CML cell lines initially induced phosphorylation of Jun kinase. We next examined the expression of heat shock proteins (Hsp70, Hsp90) after treating for 6, 12, 24 hours with the same proteasome inhibitors. Western blot analysis results indicated that expression patterns were different between MG115 and MG132. MG115 induced the slight increase of Hsp70 and Hsp90 in K562, but the reduction of both in KCL22. Meanwhile, MG132 produced the decrease of Hsp70 and Hsp90 in both K562, KCL22. In summary, our work supports that a proteasome inhibitor can induce apoptosis in human CML cell lines. We are currently focusing on the combined effect of proteasome inhibitor and Hsp90 inhibitor on CML.

IC50 of Proteasome Inhibitors

Cell line Proteasome Inhibitor 24hr 48hr 72hr K562 MG115 3.01 μM 1.14 μM 0.59 μM K562 MG132 μ 2.13 M 1.03 μM 0.57 μM KCL22 MG115 156.92 μM 1.36 μM 0.73 μM KCL22 MG132 1.56 μM 0.93 μM μ 0.75 M
A New Abl Kinase Inhibitor (AMN107) Has In Vitro Activity on CML Ph+Blast Cells Resistant to Imatinib.
Authors: Martinelli Giovanni , Martelli Alberto M. , Grafone Tiziana , Rosti Gianantonio , Mantovani Irina , Cappellini Alessandra , Tazzari Pier Luigi , Ottaviani Emanuela , Renzulli Matteo , Amabile Marilina , Poerio Angela , Soverini Simona , Terragna Carolina , Taccioli Cristian , Bassi Simona , Baccarani Michele
Published: April 2020

Imatinib mesylate (STI571), an inhibitor of the bcr/abl tyrosine kinase, is rapidly becoming the first-line therapy for chronic myeloid leukemia (CML). Imatinib has proved remarkably effective at reducing the number of leukemia cells in individual CML patients and promises to prolong life substantially in comparison with earlier treatments. However, the development of resistance to this drug is a frequent setback, particularly in patients in advanced phases of the disease. Therefore, new inhibitors of bcr/abl are needed. Very recently, a new bcr/abl inhibitor, AMN107, has been synthesized. We have tested AMN107 on leukemic cell lines and on blasts isolated from imatinib-resistant CML patients. Western blot analysis with phosphospecific antibodies revealed that in K562 cells AMN107 (10 nM) down-regulated phosphorylation of bcr/abl Tyr177, while the phosphorylation levels of Tyr412 were unaffected. This finding seems particularly important because recent evidence has demonstrated that the signaling pathway emanating from Tyr177 plays a major role in the pathogenesis of CML. Indeed, phosphorylated Tyr 177 forms a high-affinity binding site for the SH2 domain of the adapter protein Grb2. The main effectors of Brb2 are Sos and Ras, however Grb2 also recruits the scaffolding adapter protein Gab2 to bcr/abl via a Grb2,Gab2 complex. Which results in activation of the PI3K/Akt and Erk signalling networks.

In contrast, STI571, even if used at 200 nM, did not diminish phosphorylation of bcr/abl Tyr177. At 10 nM AMN107 blocked K562 cells in the G1 phase of the cell cycle. To obtain the same effect with imatinib, a 200 nM concentration was required. AMN107 did not affect cell cycle progression of bcr/abl-negative cell lines such as HL60 and NB4, even if the concentration was raised to 200 nM. AMN107 (5 μM for 24 h) significantly increased apoptosis rate in CML blasts isolated from patients resistant to the same concentration of imatinib. Therefore, AMN107 might represent a new bcr/abl selective inhibitor useful for overcoming imatinib resistance.
Two Pregnancies Resulting in a Healthy Newborn in a CML Patient Treated with Imatinib.
Authors: Sotiropoulos Damianos , Adamidou Despoina , Athanasiadou Anastasia , Goulis Dimitrios G. , Fassas Athanasios , Anagnostopoulos Achilles
Published: April 2020

Imatinib mesylate is the new molecular targeted therapy for Chronic Myeloid Leukemia (CML). Data on fertility and pregnancy are very limited. We report the case of a 27-year old female patient having two pregnancies during therapy with imatinib. CML was diagnosed in December 2001. At diagnosis the disease was in chronic phase with 100% Philadelphia-positive metaphases in bone marrow (BM). She started therapy with IFN/Ara-C and six months later she was still with 100% Ph-positive metaphases. As she had no related transplant donor, she was started on imatinib 400mg/day. Three months later, while in complete hematologic response (CHR), she became pregnant and despite doctor’s advice, she decided not to terminate the pregnancy. At that time she had 54% Ph-positive metaphases in BM. She continued receiving imatinib during the pregnancy without any adverse effects. Ultra-sound examination revealed two normal embryos and amniocentesis yielded normal karyotypes (male and female). In 24th week of gestation she was in CHR with 20% Ph-positive metaphases in BM. Two weeks later (26th week) she delivered two newborns who both died 48 hours later due to prematurity. Laboratory tests showed no abnormalities and all parameters were normal according to their age. Seven months later and while in CHR and complete cytogenetic remission (CCR) but being BCR-ABL positive, the patient became pregnant. Once again, she continued imatinib without any adverse effects. Ultrasound parameters were normal and amniocentesis showed a normal female karyotype. After 37 weeks of gestation she delivered a healthy female newborn. Two months postpartum the child is healthy, with normal laboratory tests. The patient is in CHR, CCR and BCR-ABL positive. In conclusion we report, the case of a woman with CML in whom fertility was not affected by imatinib. In addition, Imatinib caused no abnormalities to the embryos in both pregnancies and pregnancies did not affect the therapeutic effect of imatinib. However, it is under question whether imatinib has any implication in the preterm birth of the first pregnancy.
Expression of Core-Binding Factor (CBF) Fusion Genes in Clonal Evolution of Chronic Myeloid Leukemia (CML).
Authors: Ravandi-Kashani Farhad , Kantarjian Hagop , Faderl Stefan , O’Brien Susan , Rios Mary B. , Cortes Jorge
Published: April 2020

A “2-hit” model of leukemogenesis has been proposed in which one class of mutations confers a proliferative or survival advantage to the cells and the second class serves primarily to interfere with hematopoietic cell differentiation. In support of this, FLT3 receptor mutations have been frequently reported in patients with t(8;21) and inv(16) acute myeloid leukemia (AML), otherwise known as core-binding factor (CBF) leukemias, whose fusion gene products (AML1-ETO and CBFB-MYH11) contribute to impaired differentiation of leukemic cells. In CML, enhanced kinase activity of BCR-ABL confers a proliferative and survival advantage to the leukemic cells and clonal evolution is a common event at the time of disease acceleration. However, the acquisition of CBF fusion genes has not been commonly reported during the clonal evolution of CML. We report 4 patients with CML who developed CBF type rearrangements [inv(16)(n=2) and t(8;21)(n=2)] at the time of disease progression. Patient #1, a 61 year old female, presented with myeloid blast phase disease with 46,XX,t(9;22)(q34;q11.2),inv(16)(p13q22) and was treated with imatinib achieving a hematological but not cytogenetic response. Patient #2, a 48 year old male, presented with chronic phase disease and received imatinib for 2 years, achieved a complete cytogenetic remission (CG CR) but progressed to blast phase with development of 46,XY,t(9;22)(q34;q11.2),inv(16)(p13q22). Both patients had elevated and abnormal marrow eosinophils at the time of clonal evolution. Patient #3, a 54 year old female, presented in chronic phase, received imatinib and achieved CG CR after 3 months. Blast transformation occurred after one year with 48, XX,+8,t(8;21)(q22;q22),t(9;22;19;10)(q34;q11;p13.1;q22),+der(22)t(9;22;19;10). Patient #4, a 47 year old male, presented with an extramedullary myeloid mass on his arm and features of chronic phase in the marrow examination. He was treated with troxacitabine with resolution of the mass. He was then treated with imatinib. Ten months later he developed a recurrent mass with cytogenetic studies of both the mass and marrow showing 47,XY,+8,t(8;21)(q22;q22),del(9)(q13q32),t(9;22)(q34;q11.2). To our knowledge, eleven other patients with CML with inv(16)(n=10) or with t(8;21)(n=1) have been previously reported in the literature, none treated with imatinib. Patients with inv(16) had features of AML with eosinophilia (FAB M4Eo) demonstrating dysplastic eosinophils in the bone marrow examination. Development of the CBF rearrangement was invariably associated with disease progression into the myeloid blast phase with the exception of one patient, reported to develop lymphoid blast phase, based on surface markers. CBF rearrngements occur rarely at the time of disease progression in CML and may contribute to disease transformation based on the “2-hit” hypothesis for leukemogenesis.
Flow Cytometry Applications in the Diagnosis of Myelodysplastic Syndrome (MDS): Analysis of 45 Cases.
Authors: Font Patricia , Subira Dolores , Martinez-Chamorro Maria C. , Castañon Susana , Lopez-Pascual Juan , Alonso Arancha , Perez-Saenz Maria A. , Gil-Fernandez Juan J. , Alaez Concepcion , Escudero Antonio , Tomas Jose F. , Fernandez-Rañada Jose M.
Published: April 2020

Flow cytometry is a widely used method to study most of the hematologic malignancies. The utility of bone marrow immunophenotyping for the evaluation of patients with myelodisplastic syndrome (MDS) is currently being evaluated but most of these studies are based on the analysis of a large number of monoclonal antibodies.

OBJECTIVE: To explore the potential contribution of flow cytometry to the diagnosis of MDS using a reduced panel of conjugated monoclonal antibodies (MoAb).

PATIENTS AND METHODS: 45 bone marrow specimens from patients with a diagnosis of MDS based on morphologic and cytogenetic parameters (17 RA, 12 RARS, 5 MMCL, 9 RAEB, 2 RAEB-t), were analyzed by flow cytometry. In addition, 25 samples of bone marrow obtained from patients with cytopenias, but no diagnosis of MDS, were also studied. The panel of MoAb used was designed to identify abnormalities in the differentiation pathways of erithroid (CD71 FITC/Gly A PE/CD45 PC5) and myeloid lineage (CD 16 FITC/CD11b PE/CD13 PC5) as well as the presence of specific aberrant features in the CD34+ myeloid cell population (TdT FITC/CD7 PE/CD34 PC5). All samples were analized by two independet observers. To establish the diagnosis of MDS by flow cytometry, it was necessary to describe either immunophenotypic abnormalities in both myeloid and erithroid lineages or abnormalities in only one lineage plus description of more than 5% of CD34+ cells or abnormalities in one lineage plus description of aberrant features in the CD34+ population, regardless of its percentage.

RESULTS: In 43 of the 45 samples analyzed, flow cytometric criteria of MDS were described. Only two cases (1 RARS with normal karyotype and 1 RA with complex cytogenetics) were considered normal according to the immunophenotypic criteria (95% sensivity). Regarding the cohort control, 4 samples had flow cytometric criteria of MDS; 11 samples showed isolated antigenic aberrancies in the erithroid differentiation and the 11 samples left were considered as normal.

CONCLUSIONS: Flow cytometry may be a useful tool in the diagnosis of MDS, even analysing only two hemopoietic cell lineages. The reduced panel of MoAb described here can be widely used, is easy to be applied and shows a high sensitivity and a low cost. However, the finding of isolated immunophenotypic abnormalities in some patients without cytologic data of MDS seems to negatively influence the specificity of the technique. Nevertheless, those cases presenting an abnormal immunophenotype and normal morphology should be carefully monitored during their ulterior follow-up.
Phase II Trial of Darbepoetin Alfa in Myelodysplastic Syndrome (MDS): Preliminary Efficacy, Safety, and In Vitro Results.
Authors: Gotlib Jason , Quesada Sylvia , Bhamidipati Jyoti , Chaudry Kamran , Sridhar Kunju , Dugan Kathleen , Stein Richard , Tupper Germaine , Greenberg Peter L.
Published: April 2020

Recombinant human erythropoietin (EPO) has been evaluated in MDS using a spectrum of doses and schedules, usually daily or thrice weekly, with an average erythroid response rate of 20% in published trials. Darbepoetin alfa (DA) is a hyperglycosylated analog of EPO with a longer serum half-life and greater in vivo potency. We evaluated the efficacy and safety profile of weekly subcutaneous DA dosing in MDS patients (pts) who had not received prior EPO treatment, with a hemoglobin (Hb) ≤ 10g/dL and/or red blood cell transfusion-dependence. During the induction phase, DA was begun at 4.5 mcg/kg/week (wk). For pts who did not achieve a major erythroid response by 6 wks (International Working Group response criteria), the DA dose was doubled to 9 mcg/kg/wk for 6 wks. In patients without a major erythroid response, G-CSF 2.5 mcg/kg twice weekly was added to DA 9 mcg/kg/wk for an additional 6 wks. Patients achieving an erythroid response underwent a ≥ 3-month maintenance phase. Dose reduction occurred for ≥ grade 3 non-hematologic toxicity or excess Hb response (Hb ≥ 13 g/dL). To date, 12 pts have been enrolled. FAB diagnoses were: RA (n=3), RARS (n=5), CMML (n=1), RAEB (n=3); divided into IPSS Low (n=6), Intermediate-1 (n=4) and Intermediate-2 (n=2). Of the 10 pts evaluable for erythroid responses (2 were too early), 7 responded: 5 major (3 RA, 2 RARS), 2 minor (RARS, RAEB), with 3 non-responders (2 RAEB, 1 RARS). The median durability of the major erythroid responses was 12.5 months. Doses for the 5 major erythroid responders were: DA 4.5 mcg/kg/wk (1), DA 9 mcg/kg/wk (1), and DA 9 mcg/kg/wk + G-CSF (3). DA was dose-reduced in 2 patients for excess Hb response. DA was generally well tolerated with no unexpected non-hematologic toxicity. One pt with baseline diabetic chronic renal insufficiency developed grade 2 proteinuria and increased creatinine levels, and grade 3 hypertension. Of 3 patients hospitalized for pneumonia, 1 (RAEB) expired. Progressive thrombocytopenia (grade 2 to 3 or worsening grade 3) occurred in 3 pts (2 RAEB, 1 RARS). Evaluation of the stimulatory effects of DA and EPO on in vitro marrow erythroid colony formation indicated that for those MDS pts with BFU-E growth (4 of 7 pts), similar BFU-E efficacy occurred compared with normal marrow (n=7), with DA being less stimulatory than EPO. These findings are consistent with known differences between in vitro responses (a function of receptor binding affinities) and in vivo efficacy (related to the extended serum half life of DA compared with EPO). Plateau (peak)/2 stimulatory concentrations were: DA 1nM (37ng/ml), EPO 0.2nM (0.78U/ml). In vivo/in vitro correlations are being assessed. These initial data indicate good tolerability and encouraging erythroid responses with once weekly DA dosing with or without G-CSF in MDS pts. Updated results of ongoing accrual will be presented.
Clinical Course and Prognosis in Mast Cell Proliferative Disorders: A 17 Year Experience in a Single Center.
Authors: Sperr Wolfgang R. , Wimazal Friedrich , Boehm Alexandra , Hauswirth Alexander W. , Florian Stefan , Fritsche-Polanz Robert , Foedinger Manuela , Valent Peter
Published: April 2020

In patients (pts) with mast cell (MC) proliferative disorders, the clinical course, prognosis, and outcome vary depending on age, organ-involvement, and the disease-variant. We have retrospectively analyzed the clinical course and outcome in 56 pts with MC disorders refered to the University of Vienna between 1987 and 2004. Using WHO criteria, pts were found to have cutaneous mastocytosis (CM, n=6), indolent systemic mastocytosis (ISM, n=35), aggressive SM (ASM, n=3), mast cell leukaemia (MCL, n=1), SM with associated clonal hematologic non-MC-lineage disease (SM-AHNMD, n=6), and myelomastocytic leukemia (MML, n=5). These groups differed from each other in serum tryptase levels, hemoglobin, platelet counts, and lactate dehydrogenase (p<0.05). By contrast, no significant differences were found in white blood cells, eosinophils, monocytes, or presence of the c-kit mutation D816V. Most pts with ISM showed a stable clinical course without signs of progression or increase in serum tryptase even when monitored for more than a decade. By contrast, in all pts with ASM, MCL, SM-AHNMD, and MML, the neoplasm progressed after a latency period of several months. The median event-free survival was 3.7 months in pts with ASM/MCL, 22.8 months in SM-AHNMD, and 5.0 months in MML. In CM and ISM, the median survival was not reached. The differences in event-free survival among WHO groups were found to be significant (p<0.05). Apart from high grade WHO categories, adverse prognostic variables concerning survival were i. absence of skin-lesions, ii. high percentage (>5%) of MC in bone marrow smears, and iii. splenomegaly. Polychemotherapy resulted in complete remission in MML, but not in ASM or MCL. Interferon-alpha and 2CdA showed cytoreductive effects in some pts with ASM, but did not work in MCL. For the latter group of pts new therapies need to be developed.