bluuebunny/crossref_metadata_2025 huggingface dataset:
Outcome of Diffuse Large B Cell Lymphoma in the VA Health System: Do We Need To Do Better?.
Authors: Komrokji Rami S. , Maraboyina Sanjay , Safa Malek M. , Nahleh Zeina A. , Haddad Rami Y.
Published: April 2020

Background: Diffuse Large B cell lymphoma (DLBCL) is the commonest subtype of non-Hodgkin Lymphoma (NHL). Data on outcome of DLBCL in large cohorts of patients or community practices outside clinical trials are lacking. Treatment and outcomes of DLBCL among the VA patients have not been reported previously. We utilized the VA Central Cancer Registry to study cases of DLBCL in the VA system.

Methods: This was a retrospective analysis. The VA Central Cancer Registry (VACCR) database was used to identify patients with DLBCL diagnosed between 1995 and 2005. There are approximately 120 VA medical centers diagnosing and/or treating patients with cancer. The VACCR aggregates the data collected by the medical centers’ cancer registries. Data were extrapolated and analyzed using bio-statistical software SPSS. Variables included age, sex, stage of disease, histology subtype, date of diagnosis, date of last contact, date of relapse, vital status, whether patients received chemotherapy and or radiation.

Results: There were 2792 patients with DLBCL at the VACCR between 1995 and 2005, 2402 were white, 323 black patients, and 67 patients from other racial groups. The mean age at diagnosis was to 65. Majority were males (2721 patients). More patients were diagnosed at advanced stage (stage III and IV) (40%), in 34% of cases stage was not known or missing. IPI score data were not available. Most patients received multi-agent chemotherapy 70%, and 20% of patients received radiation (31 % in early stage and 12% in late stage). The 5-year overall survival was 28%. 5 year OS was 41% for stage I, 32% for stage II, 25% for stage III and 17% for stage IV. Using Cox regression multivariable analysis age and use of mutli-agent chemotherapy were the only statistically significant independent variables affecting survival.

Conclusions: Outcome of DLBCL in VA patients seems inferior to what is reported in clinical studies. The age of patients treated in the VA as well as sex could be factors affecting the outcome. There are no data available on outcome of DLBCL in such large cohort of patients outside context of clinical trials to make meaningful comparisons.
Fludarabine and Mitoxantrone for the Refractory Relapse Treatment of B-Cell Low-Grade Non-Hodgkin Lymphoma (NHL): LACOHG First Interim Report (Latin American Cooperative Oncology Hematology Group).
Authors: Baltazar-Arellano Severiano , Pimentel Patricia , Vera Luis , Bezares Fernando , Málaga Jose , Huamani-Z Julia , Montante Antonio , Rivas-Vera Silvia , Carrasco-Yalan Antonio A. , Beltran-Garate Brady E.
Published: April 2020

Background: fludarabine (F) is licensed for the management of indolent non Hodgkin’s lymphoma in countries such as Canada and Switzerland. Clinical evidence suggests that fludarabine monotherapy is as least as effective, than conventional therapies such as cyclophosphamide, vincristine, prednisone (CVP) for the first and second line treatment of B-cell low grade NHL achieving objective response rates. Better response rates can be achieved combining F with Mitoxantrone (M) in low grade NHL even in refractory relapsed (RR) patients (pts). The Latin American Cooperative Oncology Hematology Group (LACOHG) proposed a multicenter study in Latin American countries in 2002 to use FM in RR B-cell low grade NHL.

Aims: to assess the response rate, safety, disease free survival (DFS) and overall survival (OS) of FM in RR B-cell low grade NHL during (2003–2006).

Methods: Forty-eight patients in the period of January 2003 to February 2006 were evaluated. Forty-four pts. had follicular lymphoma and 4 small lymphocytic lymphoma. Median age 63.5 years old (range: 24–83). Gender: female 56% and male 44%. Inclusion criteria for low grade NHL-LG was: any previous treatment excluding autologous transplantation, Ann Arbor stage II to IV, age > 18 years old, ECOG performance status 0–2 and written informed consent. ECOG performance status 0: 2%, 1: 71% and 2: 27%. Ann Arbor staging: II: 2%, III: 29% and IV: 69%. International Prognostic Index (IPI): 0–1: 19%, 2–3: 71% and 4–5: 10%. Median previous treatment was 1 (range: 1–3). FM treatment consisted of F 25 mg/m2 i.v. (day 1–3) and M 10 mg/m m2 i.v. (day 1) each 28 days for 6–8 cycles.

Results: on this low grade NHL cohort the overall response rate (ORR was 81%; progressive disease and non-response 19%. With a median follow up of 17 months, OS at 24 months was 86% (DE 5.2%) and DFS at 24 months 57.1% (DE 11.3%). LDH in serum was not an adverse prognostic factor for DFS and OS.

Safety: on the 286 cycles in 48 pts, the toxicity was: 18 episodes of grade 3-4 neutropenias, 15 episodes of grade 3-4 thrombocytopenia, 7 episodes of grade 1–2 nausea/vomiting, grade 1–2 diarrhea in 4 pts, 8 pts were admitted to the clinic, 11 fever episodes, 2 allopecia, 4 pts developed grade 1–2 peripheral neuropathy and infections 7%: one case herpes zoster.

Mortality rate: 12,5% (6/48 patients), 5 of them because progressive disease. No cardiac toxicity was reported.

Conclusions: FM is an effective and safe treatment for RR low grade NHL. A longer follow up and a larger trial, might be needed to confirm these results in a multicenter, randomized study.

DFS with FM in RR low grade NHL : LACOHG DFS with FM in RR low grade NHL : LACOHG
High Expression and Persistent Activation of STAT1, 3 and 5 Predicts Vorinostat (Suberoylanilide Hydroxamic Acid, SAHA) Resistance in Cutaneous T-Cell Lymphoma Cells.
Authors: Fantin Valeria R. , Loboda Andrey , Roth Jennifer A. , Paweletz Cloud , Randolph Sophia , Pierce Jacqueline W. , Harrington Elizabeth A. , Hendrickson Ronald C. , Reilly John F. , Ware Christopher , Kadin Marshall E. , Duvic Madeleine , Frankel Stanley R. , Richon Victoria M.
Published: April 2020

The hydroxamic acid vorinostat (SAHA) is an HDAC inhibitor that induces differentiation, growth arrest and/or apoptosis of malignant cells both in vitro and in vivo, and is currently being tested in the clinic for a variety of indications. In particular, vorinostat has demonstrated an overall response rate of approximately 30% in advanced cutaneous T-cell lymphoma (CTCL). The purpose of this study was to identify biomarkers that predict response to vorinostat in CTCL using preclinical model systems based upon gene expression profiling and pathogenesis of lymphoma. Herein we report the results of our evaluation of the STAT (signal transducer and activator of transcription) signaling pathway. The relative level of mRNA and protein expression as well as activation status of several members of this family have been evaluated in lymphoid cell lines with diverse vorinostat sensitivity (proliferation/viability IC50 range: 0.3–14 mM). Among them, the more responsive CTCL cells lines HH and HUT102 exhibited an IC50 ≃ 0.8 mM. These cells underwent apoptosis in response to vorinostat as assessed by TUNEL assay, in contrast to Hut78 and MJ cells (IC50 > 2 mM) that did not show signs of DNA fragmentation upon incubation in vorinostat-containing media for 48 h. Our results indicate that in lymphoma cell lines elevated protein levels and persistent activation of STAT1, 3 and 5 correlate with resistance to vorinostat. Immunofluorescence microscopy studies revealed that STAT proteins preferentially localize to the nuclear compartment in cells with impaired vorinostat response, consistent with the expected distribution for this group of transcription factors in the active state. Simultaneous treatment with a pan-JAK inhibitor (JAK inhibitor I; Calbiochem) resulted in an additive antiproliferative effect consistent with a survival, antiapoptotic role for STAT proteins in the response to vorinostat treatment. Immunohistochemical analysis of STAT1 in skin biopsies isolated from CTCL patients (N = 49) enrolled in the vorinostat Phase IIb clinical trial showed that malignant T cells were positively stained in approximately half of the samples (21 positively stained; 25 negatively stained; 3 poor quality specimens), and in those cases a relationship between nuclear accumulation of STAT1 and lack of response to treatment exists (p=0.007<0.05; Fisher’s exact test). In conclusion, our results suggest that deregulation of STAT signaling plays a role in resistance to vorinostat in CTCL and strategies that block this pathway may improve response to vorinostat.
Activity of Bortezomib in Mantle Cell Lymphoma, Diffuse Large B Cell, and Small Lymphocytic Lymphoma.
Authors: Fauble Veena S. , Lobocki Catherine , Eapen Amy , Terebelo Howard R.
Published: April 2020

Bortezomib (PS341, Velcade) is a selective, reversible proteosome inhibitor. It is a unique inhibitor of the ubiquitin-proteosome pathway leading to arrest in tumor growth, induction of apoptosis, inhibition of tumor metastasis and angiogenesis. It plays an important role in the activation of NF Kappa B, which is responsible for transcription of inhibitors in apoptosis. It is a novel therapeutic agent that has demonstrated in vitro and in vivo activity in mantle cell lymphoma (MCL). In one phase II study, O’Conner et al reported that bortezomib demonstrated a response rate of 50% in refractory cases of mantle cell lymphoma, where as in small lymphocytic lymphoma there was a 0% response rate. Why certain types of non-Hodgkins lymphoma (NHL) are sensitive to this agent and others resistant is currently not well understood. It is believed that disregulation has been shown to play a role in the development of drug resistance in NHL Our objective was to determine the efficacy of bortezomib in vitro using three subtypes of NHL. Three lymphocytic cell lines were used: Granta-519 (mantle cell lymphoma), Toledo (diffuse large cell lymphoma) and EHEB (chronic B cell leukemia). The cytotoxic effects were determined by the colorimetric MTT assay after 24 hours of treatment with bortezomib (1 – 20 nM). Apoptosis was assessed by measuring Caspase-3 activity after 6 and 16 hours of treatment with bortezomib (4 or 20 nM). Bortezomib demonstrated a dose-dependent cytotoxic effect for all three cell lines, with an IC50 of 3.5 nM, 4.1 nM, and 18.5 nM for Toledo, Granta-519 and EHEB, respectively. Apoptotic studies showed that bortezomib (20 nM) induced a 1.4 to 5.6-fold increase (after 6 hrs) and 5.8 to 16.6-fold increase (after 16 hrs) in apoptosis. The EHEB cell line was most resistant, showing no increase in Caspase-3 activity at the lower dose (4 nM) at either time-point. Granta-519 was most sensitive showing a 5.2-fold increase in apoptosis with the lower dose after 16 hours of treatment. This in vitro study demonstrated a dose and time dependent response of bortezomib in both the mantle cell lymphoma (Granta-519) and the diffuse large cell lymphoma (Toledo) cell lines. The resistance to bortezomib in the chronic B cell leukemia (EHEB) cell line was confirmed in this study as evidenced by the lack of Caspase-3 activity at the 4 nM dose at both 6 and 16 hours. Our future studies will attempt to unmask the reason for this resistance. Perhaps manipulation of the apoptotic pathway through the use of combinations of various biologic response modifiers may be an attractive option in overcoming resistance in certain subtypes of NHL.
First Line Treatment with the R-VAD+C Regimen Followed by ASCT for Patients under 65 with a Mantle Cell Lymphoma (MCL). From the GOELAMS Group.
Authors: Gressin Remy , Maugendre Sylvie Caulet , Damaj Ghandi , Milpied Noel-Jean , Senecal Delphine , Berger Mathilde Hunault , Yamani Abdrrazak El , Maisonneuve Herve , Tournilhac Olivier , Celigny Philippe Solal , Quitet Philippe , Feugier Pierre , Choufy Cachera , Fontan Jean , Barbe Martine Escoffre , Cornillon Jerome , Lemevel Annie , Martin Claude , Moreau Philippe , Colombat Philippe
Published: April 2020

This protocol followed the first GOELAMS protocol (also presented at ASH 2006) for MCL pts testing the addition of Rituximab with the VAD+C regimen followed by ASCT. Thirty nine pts were included in this phase II prospective study opened between January 2003 and December 2005.

Inclusion criteria: MCL by the WHO (common or blastoïd variants), under 65 years with PS under 3, who never received chemotherapy.

Treatment: the first step consist of 4 courses of R-VAD+C regimen (J1, Rituximab 375 mg/sqm; J1 to J4, Vincristin 0,4 mg/D, Adriblastin 9 mg/D and Dexamethasone 40 mg/D; J20 to J29 Chlorambucil 12 mg/D) every 35 days. Evaluation after 4 cycles with the Cheson criterias. Responders (at least 50%) have been collected after a 4 g/sqm cyclophosphamide stimulation and an in vivo purge with rituximab (375). Then, the second step consisted on one R-VAD+C and one VAD+C regimens followed by the transplantation prepared by the myeloablative regimen including Alkeran 140 mg/sqm and 8 grays/4 fr TBI.

Results: 38/39 pts are evaluable. 34 biopsies were reviewed and consisted of 26 common forms and 8 blastoïd variants. 4 other diagnosis were possible only on blood (n=3) or bone marrow (n=1) analysis.

Treatment: 28/38 (74%) have had a response to the first 4 R-VAD+C, 6 progressed (16%) and 4 (10%) have had only a minor response under 50%. 17/38pts (44,5%) were in CR or CRu after 4 R-VAD+C courses and 11 (29%) in PR>50%. Among the 23 pts evaluated after the transplantation, 20 were in CR, two progressed and one have a PR>50%.

Survival: With a median follow-up of 19 months, OS and PFS at 2 years are 80% and 50 % respectively. The PFS is significantly affected by the Goelams Index for MCL described recently from a cohort of patients studied in our previous protocol (see abstract ASH 2006).

Conclusion The R-VAD+C regimen followed by PBSCT is a good sequence for the treatment in first line of MCL patients, especially for those with a good prognosis profile defined by the new Goelams prognostic index which would worth being tested on a larger cohort.
Rituximab Maintenenance Therapy in CD20+ B-Cell Non-Hodgkin-Lymphoma - Results of a Multicenter Prospective Randomised Phase II Study.
Authors: Witzens-Harig Mathias , Hensel Manfred , Schmier Johann W. , Neben Kai , Benner Axel , Dreger Peter , Kuhn Christian , Schmidt-Wolf Ingo , Krämer Alwin , Ho Anthony D.
Published: April 2020

Clinical and pharmacokinetic data suggest that the effect of rituximab could be improved by prolonged exposure to the drug. To test for this hypothesis we performed a prospective randomized trial of rituximab maintenance therapy in patients with CD20+ B-cell Non-Hodgkins-Lymphoma. After completion of standard treatment patients were randomized to either observation or maintenance therapy with rituximab (375 mg/m2) every 3 months for 2 years. Patients after first line therapy as well as relapse patients were included in the study. Patients with aggressive lymphoma were enrolled if they had achieved a complete response (CR) after initial treatment. Patients with aggressive lymphoma with residual tumor mass were examined with positrone emission tomography (PET) and qualified for randomization if PET showed no signs of tumor activity. Patients with indolent lymphoma qualified for the study if at least a partial response (PR) was achieved. So far 162 patients (pts) with CD20+ B-cell Non-Hodgkins-Lymphoma were enrolled in this trial. Histological subtypes included diffuse large cell lymphoma (69 pts), follicular lymphoma (41 pts), mantle cell lymphoma (18 pts), primary mediastinal lymphoma (15 pts), marginal zone lymphoma (9 pts), Burkitt’s lymphoma (3 pts), immunocytoma (2 pts), primary intestinal lymphoma (1 pt), hairy cell leukemia (1 pt), chronic lymphocytic leukemia (1 pt) and unclassified B-cell lymphoma (2 pts). No severe adverse events were observed during rituximab maintenance therapy. We conclude that rituximab maintenance therapy is feasable, safe and well tolerated in patients with CD20+ B-cell Non-Hodgkins-Lymphoma. Results including event free survival and overall survival for the observation group and for the maintenance therapy group will be presented.
Rituximab-CHOP Versus CHOP Alone in Patients with Diffuse Large B Cell Lymphoma.
Authors: Cetiner Mustafa , Salepci Taflan , Atesoglu Elif Birtas , Gumus Mahmut , Guven Aslihan , Undar Levent , Tuglular Tulin Firatli , Bayik Mahmut
Published: April 2020

Recently, the cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP) regimen plus rituximab (R-CHOP) have been used widely to treat patients with newly diagnosed diffuse large B cell lymphoma (DLBCL) and it has also been reported to improve the outcome of DLBCL.

We represent a retrospective analysis of newly diagnosed DLBCL patients between the years of 2003–2005 to evaluate the impact of R-CHOP therapy on response rates.

Patients with DLBCL between 20–80 years of age (median: 46.0 and mean 56.2 ± 14.92) received 6 cycles of R-CHOP (n=28). For comparison, DLBCL patients between 15–76 years of age (median: 60.5 and mean 47.3 ± 16.6) who received 6 courses of CHOP therapy (n=30) were used as the control group. All patients received classical CHOP (cyclophosphamide 750 mg/m m2, doxorubicin 50 mg/m2, vincristine 1.4 mg/m m2 on day 1 and prednisone 40 mg/m m2 for 5 days) every 4 weeks. In R-CHOP group, rituximab 375 mg/m m2 was administered one day before CHOP chemotherapy.

The median follow-up for R-CHOP and CHOP groups were 15.66 ± 5.90 (7–29) and 21.79 ± 9.20 (8–46) months, respectively. The International Prognostic Index (IPI) scores were not significantly different between these groups (median IPI of R-CHOP: 2.0 and mean IPI 2.01.27 ± 1.16 versus median IPI of CHOP: 1.0 and mean IPI 1.88 ± 1.26).

Complete response (CR) and complete undetermined response (CuR) rate for R-CHOP was 92% (26 of 28 patients) which was statistically significantly higher than CHOP (24 of 30 patients, 80%) (p=0.004). Partial response (PR) rates for R-CHOP and CHOP groups were 7% (2 of 28 patients) and 10% (3 of 30 patients), respectively. While there were no unresponsive patients in the R-CHOP group, refractory disease rate was 10% (3 of 30 patients) in the CHOP group. Relapse rates during the follow up period were 13% (4 of 30 patients) for CHOP and 4% (1 of 28 patients) for R-CHOP group (p<.0001). No long-term toxicity appeared to be associated with the addition of rituximab to the CHOP combination.

These results also confirmed the benefit of the addition of rituximab to standard CHOP chemotherapy in DLBCL even in young patients with low IPI scores.
Safety of Intrathecal Rituximab as Prophylaxis or Treatment in CD20+ Acute Lymphoblastic and Aggressive Lymphoma: Report from a Mexican Pilot Study.
Authors: Villela Luis , Caballero-Caballero Rocio , Piedras-Ross Josefa , Garcia-Herrera Hetor , Mejia Dolores , Escoboza Jose R. Borbolla
Published: April 2020

CNS infiltration of neoplastic B-cell lymphocytes both in acute leukemia as well as in lymphoma, carry very bad prognosis. There are no universal guidelines for the prophylaxis of such CNS relapse in these patients. Current prophylactic regimes have early and late side effects, sometimes with severe toxicities. Although rituximab has improved treatment results in lymphoproliferative diseases, it cannot reach the CNS when administered systemically (Feugier, Ann Oncol 2004, Rubenstein, Blood 2004). Schulz (Haematologica 2004) used this drug in the treatment of relapse of primary brain lymphomas with positive results using a dose of 10 to 40mg, and estimated 25mg to be a good standardized dose.

After approval from our ethics committee and obtaining the informed consent signature, patients received 25 mg of intrathecal rituximab (IT-R) according to the standard intrathecal regimen, plus the standard systemic chemotherapy protocol in our department. Side effects were evaluated according to the NIC toxicity scale.

Nine patients have been enrolled in the protocol so far: 8 were male and 1 female, median of age was 49 years (range 14–72). A total of 57 IT-R were administered to our patients (median: 6, range 3–15). They had the following diagnosis: Two with large diffuse B cell lymphoma (LDBCL); two with mantle cell lymphoma (one of them blastic type), three with acute lymphoblastic leukemia; one with Burkitt’s lymphoma (BL) and one with anaplastic lymphoma. Two patents had initial CNS infiltration; the blastic type Mantle Cell Lymphoma (patient 2) and the anaplastic Lymphoma (patient 9); these patients received IT-R three times per week, plus two extra doses after complete remission in CNS. Median follow-up was 9 months (range: 3–15 months). All patients had risk factors that warranted prophylaxis (see table). Toxicity included headache in 35% of administrations (Grade I/II), temporal paresthesias in 24.5% of administrations (lasting a median of 5 minutes, range 2–15), neuropathic pain and vomit in 12.3% of administrations (grade I/II), fever and chills in 7% of administrations (Grade I/II). All side effects were temporary and no patient has shown neurological or other late toxicities. The patient with mantle cell lymphoma infiltrated to CNS achieved complete remission after 5 doses of IT-R which continues after autologous stem cell transplantation.

The use of rituximab as IT-R is safe and effective. We are starting a phase II study using IT rituximab as prophylaxis and treatment, in a larger group of patients.
Efficacy of Rituximab in Hairy Cell Leukemia.
Authors: Kokoris Styliani I. , Angelopoulou Maria K. , Galani Zacharoula I. , Anargyrou Konstantinos , Sachanas Sotirios , Vassilakopoulos Theodoros P. , Siakantaris Marina P. , Kontopidou Flora N. , Dimitriadou Evangelia M. , Dimopoulou Maria N. , Tsaftaridis Panayiotis , Plata Eleni , Kalpadakis Christina , Kyrtsonis Marie-Christine , Panayiotidis Panayiotis , Pangalis Gerassimos A.
Published: April 2020

BACKGROUND: Hairy Cell Leukemia (HCL) is a rare B-chronic lymphoproliferative disorder (BCLD) with an indolent course. First-line treatment modalities include 2-chlorodeoxyadenosine (2-CDA), 2-deoxycoformycin (2-DCF) and interferon-alpha. The efficacy of anti-CD20-Rituximab (R) in other BCLDs, as well as strong CD20 expression by HCL cells, indicate that R could be an alternative treatment of HCL.

AIMS: The experience of a single Hematology Unit in the treatment of relapsed HCL with R.

PATIENTS AND METHODS: We retrospectively analyzed all HCL patients (pts) who received R as salvage therapy in 1st or subsequent relapse.

RESULTS: 13 patients treated with R were located among 110 patients diagnosed with HCL between 1980 and 2005. 11 were males and their median age before R treatment was 46 years (range: 42–88). 5 pts had splenomegaly with a median spleen size of 7cm below left costal margin (range:5–20cm). 3 pts has an absolute neutrophil count<1.5×109/L, 4 a hemoglobin <10g/dL and 5pts a platelet count<100×109/L. All patients displayed a typical immunophenotype from blood and/or bone marrow (CD20 strongly +, CD19+, CD22+, FMC-7+, CD11c+, CD25+, CD103+). Four of them were CD23+ and two CD10+. 8 pts received Rituximab at 1st relapse. Among them, one had received 2-DCF as first-line treatment, one 2-CDA and 6 interferon-alpha as induction and maintenance. 3 pts had received more than one prior treatments. Two pts received R at diagnosis, due to older age. The median time from diagnosis to R initiation was 61 months (range: 4–275). R was administered at 375mg/m2 weekly for 6 cycles. Overall response rate was 67%, with 4 pts showing a negative immunophenotype. One pt discontinued treatment after the first cycle due to the development of thrombocytopenia that was attributed to the drug. 7/8 responders showed a complete restoration of their cytopenias. No other complications were recorded, except of mild infusion-related symptoms. Among the responding pts, none has relapsed so far with a median follow-up of 14 months (range: 4–40+). Among partial responders, one achieved a complete response including a negative bone marrow and immunophenotype after R retreatment.

CONCLUSIONS: R is a highly effective and tolerable treatment for HCL in relapse with a response rate of 67%. Retreatment or maintenance with R may be important, since ongoing responses are seen.
Rituxan-HCVAD in Mantle Cell Lymphoma: To Transplant or Not To Transplant: The University of Pennsylvania Experience.
Authors: Nasta Sunita Dwivedy , Xavier Marin F. , Andreadis Charalambos , Tsai Donald E. , Porter David , Luger Selina , Schuster Stephen J.
Published: April 2020

Mantle cell lymphoma is a difficult disease entity with no standard therapy. The best reported results for treatment of de novo patients has been from the MD Anderson Cancer Center with long term remission rates reported of 77%, using the HCVAD regimen in combination with autologous stem cell transplantation. (Khouri, ASH 1998) In later reports, the addition of Rituximab to the HCVAD regimen appears to obviate the need for additional consolidative auto-PBSCT. (Romaguera, ASH 2005) In a retrospective analysis of patients treated at the University of Pennsylvania between 1999 and 2006, we seek to compare patients receiving the R-HCVAD regimen without transplant to those who received auto-PBSCT in the same time period. Patients were stratified by their transplant status. Survival data, disease status, and secondary malignancy information were assessed. Twenty-five patients were identified, with an age range of 41–76 years at diagnosis. All but one patient had advanced disease at diagnosis (stage III or IV) There were 18 men and 7 women. 17 patients received R-HCVAD alone, and 8 patients went on to Cyclophosphamide/TBI autologous stem cell transplant. Survival in the patients treated with R-HCVAD alone was 82.3% with a median follow-up of 3 years, with all deaths due to progressive disease. Of surviving patients in this group, 1 out of 14 has had additional therapy but remains in second complete remission. Survival of patients post-transplant was 75%, with one death due to disease and one death due to secondary malignancy (lung cancer). One additional case of secondary MDS was also reported in the transplant group. In summary, our series reveals that the R-HCVAD therapy can provide excellent long-term remission rates with no additional survival advantage gained from autologous transplant. An increased risk for secondary malignancies was observed in patients undergoing autologous PBSCT. Further prospective studies of consolidative therapy are warranted in this population.
The Anticancer Effect of Cytochrome P450 2E1 Gene Which Was Mediated by Recombination Adenovirus in Mesenchymal Stem Cells.
Authors: Wang Jishi , Hu Xiaoyan , Liu Xinmei , Yang Yuan , Wu Yingya , Fang Qin
Published: April 2020

Dacarbazine (DTIC) can inhibit purine nucleotide synthesis after activated by P4502E1(CYP2E1) in liver and can be used for the treatment of malignant melanoma and Hodgkins lymphoma.

Objective: To explore the anticancer effect of the combination of CYP2E1 and DTIC.

Methods:

A full length cDNA fragment encoding the human CYP2E1 was cloned and was transfected into bone marrow mesenchymal stem cells (BMSCs) through recombinant adenovirus vector. The recombinant adenovirus vectors pAd5CMV-CYP2E1 and pAd5CMV-EGFP were constructed and transfected into the packing cell lines HEK293 cells by liposome encapsulation method. The expression of CYP2E1 gene was detected by Real-time PCR and Western Blot and contrasted to the cells which were transfected into the recombinant adenovirus including EGFP. In vitro, gene transfected BMSCs were co-cultured with DTIC, and then the supernatant was added into the medium of human malignant melanoma M14 cells. Fluorescence microscopy, DNA Ladder, flowcytometry, electrophoresis were employed to tests the apoptosis, and MTT method was applied to detecting the IC50. In the animal experiment, the CYP2E1 gene tranfected BMSCs were injected in combination with DTIC directly into the malignant melanomas on nude mice or through caudal vein, and the injected BMSCs transfected pAd5CMV-EGFP and DTIC in the same manner into nude control mice. The inhibiting effect of new planted tumors of experimental group was monitored through Immuno-histochemistry analysis and the shape and size of tumors were measured. CYP2E1-EGFP gene tranfected BMSCs were injected into nude mice through caudal vein and tracked by fluorescence.

Results:

CYP2E1 gene(1482bp) was cloned successfully and it was conformed by DNA sequencing; Endonuclease and PCR analyses showed that recombinant adenovirus vector had been constructed and high titer recombinant adenovirosome(1×1012pfu/ml) had been packaged successfully. BMSCs had been isolated and flowcytometry showed their phenotype were CD44(+),CD105(+), CD34(−)and CD45(−). The mRNA and protein quantities of the group of CYP2E1 gene tranfected BMSCs were significantly higher than the control group (P<0.05, n=24). In vitro, immunity fluorescence microscope, DNA Ladder, and flowcytometry all showed the supernatant of co-culture of CYP2E1 gene tranfected BMSCs and DTIC) significantly decreased the mortality of M14 cells compared with pAd5CMV-EGFP-BMSCs control group (P < 0. 05, n=24). In vivo, the sizes of the tumor in the nude mice treated with transfected BMSCs and DTIC were significantly smaller than control case and changed along with concentration. (P< 0.05, n=20). No new tumor was found in the nude mouse inoculated with M14 cells after the injection of CYP2E1 transfected BMSCs and DTIC.

Conclusion: Recombination adenovirus vector was able to express CYP2E1 in BMSCs. Gene CYP2E1 transfected BMSCs and DTIC showed anticancer activity both in vitro and vivo.
Evaluation of Novel Cell Cycle Inhibitors in Mantle Cell Lymphoma.
Authors: Park In-Woo , Reddy M.V. Ramana , Reddy E. Premkumar , Groopman Jerome E.
Published: April 2020

Signature abnormalities in the cell cycle and apoptotic pathway have been identified in mantle cell lymphoma (MCL), affording the opportunity to develop targeted therapies. Here, we report on a novel class of kinase inhibitors, styryl sulfones, that differ from prior cell cycle inhibitors in that they are not related to purines or pyrimidines. We observed that two closely related compounds, ON013100 and ON01370, altered the growth and cell cycle status of MCL lines and potently inhibited the expression of several important molecules, including CDK4, p53, MDM2, cyclin D, and cyclin B. Using both TUNEL and PARP assays, we found that these compounds caused apoptosis in MCL cells. In addition, using molecular analyses, we observed the modulation of caspase-3 activity but not the expression of Bcl family molecules in these cells. Next, we investigated the cytotoxicity of the MCL lines upon treatment with styryl sulfone compounds in combination with other currently used chemotherapeutic agents, such as doxorubicin (DOX) or vincristine (VCR). We found that the combination of DOX plus styryl sulfone or VCR plus styryl sulfone increased cytotoxicity by one log scale, compared with the single styryl sulfone compound. Thus styryl sulfones alone, or in combination with chemotherapeutic agents, present attractive opportunities for new drug development in MCL.
Abnormalities in Lipoprotein Metabolism Provide Insight into Novel Therapeutic Approaches for Waldenstrom’s Macroglobulinemia (WM).
Authors: Patterson Christopher J. , Moreau Anne Sophie , Leleu Xavier , Soumerai Jacob , Hunter Zachary , Hatjiharissi Evdoxia , Xu Lian , Treon Steven
Published: April 2020

WM is an incurable B-cell disorder characterized by the presence of lymphoplasmacytic cells in the bone marrow and an IgM monoclonal protein. The presence of hypocholesterolemia has often been observed in WM patients, though the incidence and significance has not been investigated. As part of these studies, we evaluated lipoprotein levels in 110 patients with WM. The median age for this cohort was 62, and median number of prior therapies was 1. Twenty-one (19.1%) patients were on a lipid-lowering agent (lovastatin, simvastatin, pravastatin). We observed decreased total cholesterol (<160 mg/dL), LDL (<100 mg/dL) and VLDL (<40 mg/dL) levels in 41 (37.3%), 68 (61.8%) and 90 (81.8%) patients, respectively. Among the 41 patients with a cholesterol level of <160 mg/dL, only 7 patients were on a lipid lowering agent. Importantly, median serum IgM levels which serve as a surrogate marker for WM disease burden were higher among patients with hypocholesterolemia (3440 vs. 1587 mg/dL; p=0.0004). Moreover, significantly lower levels of serum IgM (885 vs. 1960 mg/dL; p=0.004) were observed among patients on a lipid-lowering agent. Given these results, which suggested a possible disease dependency on cholesterol metabolism, as well as possible anti-tumor effect of statins, we explored the impact of two statins (lovastatin, simvastatin) on the growth and survival of BCWM.1 WM cells. The studies demonstrated that both lovastatin and simvastatin inhibited proliferation (by MTT assay), and induced apoptosis (by Annexin-PI assay) in a dose dependent manner (0.5–50 um) at concentrations that overlapped pharmacologically achievable levels. Taken together, these studies suggest that lipoprotein metabolism may be essential for WM growth and survival, and that statin agents may represent clinically useful agents for the treatment of WM.
In Vitro and In Vivo Anti Lymphoma Effect of GX15-070 in Mantle Cell Lymphoma.
Authors: Bebb Gwyn , Muzik Huong , Nguyen Sophia , Morris Don , Stewart Douglas A.
Published: April 2020

Introduction

Mantle cell lymphoma (MCL), an incurable B cell lymphoma, consistently over expresses bcl-2 despite not carrying the t(14;18). The attenuation of apoptosis by bcl-2 is thought to contribute to the malignant process and increase resistance to some cytotoxic agents. We recently demonstrated that GX15-070, a small molecular inhibitor of the BH3 binding groove of bcl-2, has activity against MCL cell lines in vitro. We set out to assess the effect of GX15-070 alone and in combination with Vincristine on the viability of MCL cells in vitro and in vivo.

Methods

3 previously characterized bcl-2 over expressing MCL cell lines (JVM-2, Hbl-2, granta) were used. Cells were grown in standard media and exposed to a range of concentrations of GX15-070 with and without Vincristine. Dose-response was assessed by measuring viability at 48 hours using the WST-1 assay. In vivo experiments were conducted on immune deficient mice in which 5×106 cells were injected in the flank then treated IV with GX15-070 (q 2days × 5 doses), Vincristine (q4 days × 3 doses) or both starting 5 days later. Tumours were measured three times weekly.

Results

All three MCL cell lines over-expressed bcl-2 by western blot. Each MCL cell line showed sensitivity to GX15-070 at a range of concentrations. The addition of GX15-070 to low dose Vincristine (10−6) caused significant growth inhibition of each MCL cell line (see table 1).

Discussion

Our results demonstrate that using GX15-070 to target bcl-2 is an effective anti neoplastic approach against MCL cell lines in vitro. In addition, our results suggest that combining Vincristine and GX15-070 is a promising strategy in treating MCL. In vivo experiments to confirm this additive activity are still ongoing and will be presented in full. Initial impressions suggest that there is a rationale for the addition of GX15-070 to current cytotoxic regimens used to treat MCL in the setting of clinical trials.

Table 1: Effect of Vincristine and GX15-070 on in vitro growth of 3 MCL cell lines Growth as % age of Control Cell Line JVM-2 HBL-2 Granta Vincristine alone (10-6 mg/ml) 92% 48% 89% GX15-070 alone (0.08 uM) 75% 76% 60% Vincristine 10-6 mg/ml and GX15-070 0.08 uM 52% 24% 52%
Treatment Outcomes for Imatinib Mesylate and Allogeneic Stem Cell Transplantation in Patients with CML Based on RQ-PCR, and the Efficacy of Dose Escalation of Imatinib Mesylate in Patients with Cytogenetic or Hematologic Resistance.
Authors: Sohn Sang Kyun , Cho Yoon Young , Ahn Byung Min , Chae Yee Soo , Kim Jong Gwang , Moon Joon Ho
Published: April 2020

This study examines the treatment outcome for imatinib mesylate (IM) and allogeneic stem cell transplantation (SCT) in patients with chronic myelogenous leukemia (CML) based on quantitative PCR data and the efficacy of a dose increment of IM in patients with cytogenetic or hematologic resistance.

A total of 60 Ph+ CML patients that received IM and 17 patients that received allogeneic SCT were enrolled in the current study. The cumulative incidence of a complete hematologic response (CHR) was estimated as 71% after 1 month and 93% after 3 months, while the cumulative incidence of a major molecular response (MMR) was estimated as 53% after 6 months and 73% after 1 year. Five (22.7%) out of 22 evaluable patients achieved MMR after 3 months of IM treatment. At 24 months, 10 patients (45.5%) achieved MMR, while 7 patients (31.8%) remained as non-responders. Thrirteen patients that exhibited a cytogenetic or hematologic resistance to the standard dose of IM were treated with an increased dose of IM. Four (80%) out of 5 patients who had low levels (10–55%) of Ph+ cells in their bone marrow at the time of cytogenetic resistance achieved a complete cytogenetic response (CCR) or MMR again with the increased dose of IM, while all the patients (n=8) who failed to respond cytogenetically or hmatologically to the increased dose of IM had 100% Ph+ cells in their bone marrow at the time of cytogenetic resistance. The Philadelphia positivity in the bone marrow indicated a cytogenetic or molecular response to an increased dose of IM (P=0.009). The overall survival rate at 3 years was 88% for the IM group at the median follow-up duration of 851 days (range, 13–3346 days) and 47% for the SCT group at the median follow-up duration of 456 days (range, 34–2455 days), respectively (P<0.001). Plus, the progression-free survival rate at 3 years was 61% for the IM and 47% for the SCT group, respectively (P=0.037). In conclusion, IM dose escalation can be regarded as an important treatment option in terms of reserving other treatment options, especially in patients with low levels (≤55%) of Ph+ cells in their marrow who are resistant cytogenetically or hematologically to the standard dose of IM.
Chronic Myeloid Leukemia in Two Patients with Gastrointestinal Stromal Tumor.
Authors: Sakamoto Erina , Nakamae Hirohisa , Koh Ki Ryang , Yamane Takahisa , Hino Masayuki
Published: April 2020

Imatinib competitively binds to the ATP binding site of Bcr-Abl kinase and inhibits Abl tyrosine kinase activity. Most patients with chronic myeloid leukemia (CML) achieve clinically relevant hematologic and cytogenetic responses to imatinib. It is also a potent inhibitor of a receptor-type c-Kit tyrosine kinase. Therefore, it was examined for therapeutic efficacy against malignant Gastrointestinal Stromal Tumor (GIST), which is a rare tumor of the gastrointestinal tract and is mainly caused by aberrant expression of a mutated c-Kit that is constitutively active without binding of a ligand, stem cell factor. We report two rare cases of double cancer involving CML and GIST. The first case is a 66-year-old Japanese male patient who was presented to our hospital with leukothrombocytosis on February 27, 2006. The patient was diagnosed to be suffering from CML by the presence of Philadelphia chromosome. Several days later, he was admitted with acute peritonitis. A ruptured tumor on 80 cm oral side of ileum end was found by an emergency partial ilectomy. Microscopically, spindle-shaped and round epithelioid cells were growing externally from the intestinal muscular propia. Immunohistochemical analysis showed that the tumor cells were positive for c-kit and CD34 and negative for S-100 protein and desmin. The tumors were identified as GIST. Imatinib was administered to treat CML on the 8th postoperative day. The second case is a 57-year-old Japanese female patient who was presented to our hospital with marked thrombocytosis on March 6, 2006. The patient was diagnosed to be suffering from CML by the presence of Philadelphia chromosome. One year ago, she underwent distal gastrectomy for submucosal tumor. Pathological examination revealed proliferation of submucosal spindle-shaped cells with elongated cigar-shaped nuclei arranged in fascicles. Immunohistochemical stains confirmed the diagnosis of GIST.

These two cases provided a valuable opportunity for treating GIST with CML. We look forward to active effect of imatinib, not only in CML treatment but also in preventing recurrence of GIST. The occurrence mechanism of either CML or GIST at the biomolecular level has been established. However, we can show no relation between these malignancies. Therefore, it is natural for us to understand accidental coincidence. We should follow-up these patients carefully. Accumulation of such cases may contribute to clarify more precise mechanism of tumorgenesis.
The Recent Thymic Output Function Might Relate with BCR-ABL mRNA Level in Patients with CML.
Authors: Geng Suxia , Du Xin , Weng Jianyu , Yang Lijian , Chen Shaohua , Li Yangqiu
Published: April 2020

Objective In order to analyze the relationship between the content of T-cell receptor excision DNA circles (TRECs) and BCR-ABL mRNA levels, evaluating the prognostic significance of thymic recent output function monitoring in patients with chronic myelogenous leukemia (CML).

Methods Quantitative detection of TRECs and BCR-ABL fusion gene transcripts in peripheral blood from 15 CML patients were preformed using real-time PCR technique. And the TREC-number was related to the number of T-cells by determination of the number of CD3-positive cells. The change of BCR-ABL level in 6 CML patients was followed-up for two years.

Results There was no significant correlation between TRECs and BCR-ABL mRNA in peripheral blood from CML patients at the first diagnose. Patients who had higher TRECs at diagnosis had a larger reduction of BCR-ABL level after 2 years of follow-up. While in 2 patients who underwent haemopoietic stem cell transplantation(HSCT), BCR-ABL in one patient with higher pre-transplantation TRECs value became undetectable with three consecutive detections during the first year post transplantation, but low level of BCR-ABL could be identified in the other patient.

Conclusion High thymic output function in CML patients could be helpful for anti-residual CML cells.
Single and Multiple Oral Doses of AKR-501 (YM477) Increase the Platelet Count in Healthy Volunteers.
Authors: Desjardins Robert E. , Tempel Donna L. , Lucek Rudolph , Kuter David J.
Published: April 2020

AKR-501 is an orally active thrombopoietin (TPO) receptor agonist that increases platelet production through stimulation of megakaryocyte proliferation and differentiation. The effect of AKR-501 in in vitro and in vivo models is additive with that of rhTPO. AKR-501 is a potential new therapy for use in the prevention or treatment of thrombocytopenia of various etiologies. We evaluated the safety, pharmacokinetics and pharmacodynamic activity of AKR-501 when administered as single and multiple oral doses to healthy volunteers. In the single dose study, 7 dose cohorts of 9 subjects each were randomized in a ratio of 2:1 to receive increasing doses (1, 3, 10, 20, 50, 75 and 100 mg) of AKR-501 or placebo. In the multiple dose study, 5 dose cohorts of 9 subjects each were to be randomized in a ratio of 2:1 to receive increasing doses (3, 10, 20, 50 and 100 mg) of AKR-501 or placebo administered daily for 14 days. All subjects were evaluated for safety and pharmacodynamic activity as assessed by peripheral blood platelet counts (PBPC). Dose escalation was stopped on Day 10 of dosing in the 3rd dose cohort (20 mg/day) when 6 of the volunteers exhibited a platelet count above 500,000/mL. The protocol defined endpoint of >50% increase over baseline platelet count was achieved in 5 of 6 volunteers given a single dose of 100 mg, and in all 6 volunteers given daily doses of 10 mg for 14 days and 20 mg for 10 days. The drug was well tolerated in both the single and multiple dose studies with no serious drug-related adverse experiences reported at any dose. AKR-501 was consistently well absorbed exhibiting dose linear pharmacokinetics following single and multiple dose administration, and a serum half-life of approximately 16 hours. The mean observed Cmax following administration of single doses increased from 5.67 ng/mL at 1 mg to 388 ng/mL at 100 mg. The mean observed AUC increased from 174 ng·hr/mL at 1 mg to 11052 ng·hr/mL at 100 mg. Accumulation to steady state with multiple dosing was consistent with predictions based on a linear model and the single dose PK parameter estimates. There was a linear correlation of individual observed peak activity with corresponding Cmax (single-dose r2=0.70 p<.0001, multiple-dose r2=0.67 p<0.0001). The mean change from baseline PBPC for each dose cohort in the single-dose (Figure 1) and multiple-dose (Figure 2) studies are shown in the following graphs.

Figure 1 Figure 1. Figure 2 Figure 2.
Study on the Mechanisms of CML Blast Crisis by Comparative Proteome Analysis.
Authors: Liu Xiaoli , Du Qingfeng , Zhang Song , Li Rong , Yao Feng , Fan Wei , Wang Yu
Published: April 2020

The clinical course of chronic myeloid leukemia(CML)is characteristically triphasic, comprising chronic and accelerated phases and blast crisis. Chronic phase(CP) is characterized by the Ph chromosome as the sole genetic abnormality and blast crisis(BC), which is the terminal phase of CML, often associated with additional chromosomal and molecular secondary changes. Although CML is probably the most extensively studied human malignancy, the mechanisms of CML blast crisis are still poorly understood. In current study, we analyzed the changes of protein expression between CML-CP(25 cases) and CML-BC(20 cases) by Two-dimensional polyacrylamide gel electrophoresis(2-D PAGE). Compared with that of CML-CP, 33 proteins’ intensities of CML-BC were found to have significant difference including 23 increasing and 13 decreasing. 15 proteins were identified by peptide mass fingerprint in combination with database searching including proteasome activator complex, hnRNP, annexin A4, serine proteinase inhibitor, annexin A1, GAPDH, RhoGDI, enolase, proteasome subunit 6a, GTP binding protein, leukotriene A4 hydrolase, Rac-RhoGDI, thioredoxin, proteasome subunit 4β and DJ-1 protein, and the functions of these proteins involve cell signal transduction, apoptosis, proliferation and transcription. In conclusion, our study provided a profile of protein expression difference between CML-CP and CML-BC and contributed to understand the mechanisms of CML blast crisis.
Imatinib (Glivec) and Nilotinib (AMN107), Two Selective Inhibitors of Bcr-Abl Tyrosine Kinase, Induce Hemoglobin Production in Human K-562 CML Erythroleukemia Cells in Addition To Promoting Apoptosis.
Authors: Tsiftsoglou Asterios S. , Manley Paul W. , Bonovolias Ioannis D.
Published: April 2020

Imatinib (Glivec) and Nilotinib (AMN107) have been developed as selective targeted inhibitors of Bcr-Abl chimaeric tyrosine kinase activated in CML and other malignancies characterized by the t(9;22) translocation (Manley P.W., et al. Biochim Biophys Acta 1754(1–2):3–13, 2005). Both agents are considered innovative targeted cell death promoting agents designed for treatment of human leukemias. During the course of their evaluation as apoptosis-promoting agents in human K-562 leukemia cells, we observed that both agents in addition to blocking Bcr-Abl tyrosine kinase, promoted production of hemoglobin but to different extent. Imatinib caused hemoglobin production in 6–7% of cells at concentration of 0.01 μM to 0.5 μM. This proportion reached 54% in the presence of 90 μM of Hemin. Nilotinib alone, on the other hand, caused hemoglobin production in 66% of cells at a concentration of 0.01 μM. This proportion increased to 75% in the presence of 30 μM of Hemin. These findings indicate that:

Imatinib and Nilotinib are weak and potent inducers of hemoglobin biosynthesis, respectively. Treatment of K-562 cells with either Imatinib (0.5 μM) and Nilotinib (0.01 μM) enhanced the ability of Hemin to promote erythroid differentiation. This ability of Nilotinib to induce hemoglobin accumulation at concentrations lower than those causing apoptosis, suggest that this agent may be clinically useful by promoting hemoglobin biosynthesis in diseases characterized by hemoglobin deficiency (hemoglobinopathies). We are currently investigating the mechanism whereby Nilotinib can selective activate the globin gene expression in K-562 leukemia cells.