bluuebunny/crossref_metadata_2025_split huggingface dataset:
Anti-Tumor Effect of Direct Intratumor Injection of Recombinant Fowlpox Virus Encoding CD40 Ligand in B Cell Lymphoma Model.
Authors: Liu Aichun , Guardino Alice , Panicadi Dennis , Chinsangaram Jarasvech , Levy Ronald 
Published: April 2020

Abstract
The interaction between CD40 and CD40 ligand (CD40L) on antigen presenting cells (APC) is critical in promoting cellular immune responses. In vitro experiments have shown that lymphoma B cells are efficiently infected by virus vectors encoding CD40L and subsequently up-regulate co-stimulatory molecules. By infection with CD40L, lymphoma B cells could function as APC. Tumor cells modified ex vivo to express CD40L have been shown to be able to induce protective immunity against subsequent tumor challenge. In order to develop an immunotherapy approach against established tumor, we tested the effect of intratumor injection of the recombinant fowlpox virus encoding murine CD40L (rF-mCD40L) together with systemic administration of cyclophosphamide (CTX). Methods: BALB/C mice were first inoculated s.c. with 1×107 syngeneic A20 lymphoma cells. On day 15, when tumor reached approximately 1.0 cm in diameter, mice were separated into 6 groups of ten mice which received: 1) rF-mCD40L alone, 2) empty fowlpox virus (FP-virus) alone, 3) PBS, 4) CTX (100mg/kg, i.p), 5) rF-mCD40L plus CTX, 6) FP-virus plus CTX. Virus was administrated twice by intratumoral injection at a dose of 1×109 pfu at a 7 days interval. CTX was delivered i.p at a dose of 100mg/kg on two successive days between the two virus injections. Tumor growth and animal survival were monitored every three days. In addition, tumor cell transgene expression was measured by flow cytometry 24 hours following virus injection. Results: After 24 hours of intratumor injection, a small percentage of tumor cells shown to be positive for the transgene on analysis. As shown in Figure 1, Chemotherapy alone caused only temporary regression of tumor, but the combination of rF-mCD40L with CTX resulted in tumor free mice 80 days after tumor inoculation. By contrast, mice treated with rF-mCD40L alone or with empty virus, died 4 to 7 weeks after tumor inoculation. 40 to 50% of mice treated with CTX alone or with FP virus combined with CTX exhibited re-occurrence of tumor growth and 3 mice in group of CTX alone, 2 mice in the group of FP virus combined with CTX died of tumor progression. Conclusion: rF-mCD40L alone had no anti-tumor effect against established lymphoma. Intratumor injection of rF-mCD40L in combination with systemic chemotherapy was effective for the therapy of advanced lymphoma. Intratumor administration of CD40L in combination with chemotherapy may be a promising strategy for the treatment of advanced lymphoma.
Figure Figure
The PKCβ Selective Inhibitor, Enzastaurin (LY317615), Inhibits Growth of Human Lymphoma Cells.
Authors: Rossi Randall M. , Henn Alicia D. , Conkling Rebecca , Guzman Monica L. , Bushnell Timothy , Harvey Jennifer , Fisher Richard I. , Jordan Craig T. 
Published: April 2020

Abstract
Activation of PKC contributes to tumor cell survival and proliferation and has been implicated in the pathogenesis of human B cell lymphomas. Specifically, PKCβ activation is increased in tumor cells from patients with poor prognosis diffuse large cell lymphoma (DLCL), suggesting that PKCβ may be a target for therapeutic intervention. In this study we tested the PKCβ inhibitor, Enzastaurin (LY317615), using both cell culture and xenograft assays, on a panel of eight DLCL lines (OCI-Ly3, OCI-Ly7, OCI-Ly10, OCI-Ly19, SUDHL-4, SUDHL-6, Farage, and Toledo). In suspension culture, Enzastaurin was cytotoxic to all lines tested, with an average IC50 at 48 hours of 2.0 micromolar (range 0.5–4.0 micromolar). Molecular analyses showed that Enzastaurin treatment inhibited phosphorylation of GSK3β, a potential pharmacodynamic marker of Enzastaurin activity. Subsequent studies were performed using two subclones of the DLCL lines OCI-Ly10 and OCI-ly19, which respectively represent the ABC (activated B cell) and GC (germinal center) molecular subclasses of DLCL. Each line was engineered to express luciferase, which permits real time in vivo imaging. Upon subcutaneous transplantation into immune deficient NOD/SCID mice, both lines formed reproducible tumors. Treatment with Enzastaurin (75mg/kg, b.i.d. oral gavage) was initiated during the first 1–2 weeks post-transplant, before palpable tumors were evident, and was continued for 14 consecutive days. The Enzastaurin treatment was well tolerated, with the only overt side effect being moderate weight loss. At the end of therapy, vehicle control treated tumors had become palpable and progressed to volumes of 40–60 cubic millimeters. In contrast, Enzastaurin treated tumors (both OCI-Ly10 and OCI-Ly19) showed no observable progression, remaining detectable by luciferase imaging, but non-palpable. Collectively, these data indicate the Enzastaurin can induce death of human DLCL lines in vitro and is at least cytostatic to human DLCL in vivo.
Targeting Lymphoid Malignancies with Small Molecule Inhibitors of Pro-Survival Bcl-2 Proteins.
Authors: Roberts Andrew W. , Wei Andrew , van Delft Mark F. , Mason Kylie D. , Adams Jerry M. , Huang David C.S. 
Published: April 2020

Abstract
As overactivity of pro-survival Bcl-2 proteins promotes neoplasia and enhances resistance of malignant cells to cytotoxic therapies, a promising approach for incurable lymphoid tumors is to directly target the pro-survival proteins. One approach is to mimic their physiological antagonists, the BH3-only proteins. We initially determined which of the diverse BH3-only proteins would be optimal to mimic. The interactions of the BH3 domains of this family with a groove on the Bcl-2-like proteins have been considered promiscuous. However, we found that the interactions between eight BH3 peptides and five Bcl-2-like proteins varied over 10,000 fold in affinity, and that only certain protein pairs associate inside cells (Chen et al Mol Cell;17:393–403, 2005). Bim and Puma potently engaged all the pro-survival proteins comparably. Bad, however, bound preferentially to Bcl-2, Bcl-xL and Bcl-w. Strikingly, Noxa bound only Mcl-1 and A1. In accord with their complementary binding, Bad and Noxa cooperated to induce potent killing.
We next determined the mechanism of action of putative BH3 mimetic compounds. Because killing by the BH3-only proteins require the action of the essential cell death mediators, Bax and Bak, we initially screened putative BH3 mimetic compounds on cells genetically engineered to lack both Bax and Bak. Only those compounds that mimicked the BH3-only proteins (i.e, inert on BaxBak double null cells, but active in Bax or Bak expressing cells) were evaluated further. Surprisingly, among seven putative BH3 mimetics tested, we found that only the recently described ABT-737 (Abbott Laboratories; Oltersdorf et al, Nature435:677–81, 2005), required the pro-apoptotic protein Bax or Bak to induce apoptosis. The cytotoxicity of ABT-737 alone was modest in a range of hematopoietic and non-hematopoietic cell types. Further investigations revealed that, like Bad, ABT-737 only targeted Bcl-2, Bcl-xL and Bcl-w. As hematopoietic cells typically express Mcl-1, we attempted to augment the activity of ABT-737 by concomitantly neutralizing Mcl-1. Targeting of Mcl-1 by overexpressing the BH3-only protein Noxa, or Mcl-1 down-regulation by RNAi, cytokine deprivation or cytotoxic agents, allowed ABT-737 to efficiently kill diverse cell types, even when Bcl-2 was over-expressed. We conclude that ABT-737 is a highly selective and specific BH3 mimetic compound that should prove highly efficacious in tumors where Mcl-1 is low, or when combined with agents that down-regulate Mcl-1. This hypothesis is currently being tested in vitro in primary human lymphoid malignancies, and in vivo using immunocompetent murine models of lymphoma. ABT-737 also provides strong proof-of-principle that targeting pro-survival Bcl-2 proteins is feasible, but the optimal utility of such BH3 mimetics depends on a thorough understanding of the pathways to apoptosis.
Combination of Farnesyl Transferase Inhibitor (Tipifarnib) with Perifosine Induces Apoptosis through phos-PDK1 in Human Lymphoma and Leukemia Cell Lines.
Authors: David Ebenezer , Sinha Rajni , Torre Claire , Kaufman Jonathan L. , Lonial Sagar 
Published: April 2020

Abstract
Introduction: Novel agents as anti-cancer therapy are used in the setting of specific molecular abnormalities that provide a survival advantage for malignant cells. One such agent, tipifarnib, is theoretically targeted at Ras mutations which are present in a number of different human cancers. Our previous experience with the FTIs (David et al, in press Blood) has demonstrated that they are ideal agents to combine with other targeted agents. We have investigated the combination of the AKT inhibitor perifosine with tipifarnib in human leukemia and lymphoma cell lines with the hypothesis that the combination of 2 targeted agents will disrupt separate survival pathways and ultimately result in synergistic tumor cell death.
Methods: In this study we used the human leukemia cell lines HL-60, Jurkat, and the lymphoma cell line HT. Western blot analysis was used to assess for the effect of either single agent perifosine, tipifarnib, or the combination on AKT, p-AKT, PDK-1, and caspase cleavage. Flow cytometry was utilized to assess for Annexin V staining following combination therapy.
Results:Dose escalation studies demonstrated that doses of tipifarnib up to 5μm demonstrated a significant cell death in HL-60 and HT cells. Perifosine doses of 1–5uM also induced cell death in both HL-60 and HT cells. When apoptosis was assessed using western blot analysis of caspase 3 activity and cleavage, the combination of perifosine and tipifarnib demonstrated significant apoptosis using low doses of both agents. The apoptosis was associated with downregulation of phos-PDK1, with a resultant downregulation in p-AKT. The level of phos-PDK1 was completely inhibited in less than 24 hrs in both the HL-60 and HT cell lines in combination than when either agent was given alone.
Conclusion: The combination of perifosine, and AKT targeted agent, with tipifarnib, a Ras targeted agent, appear to induce significant cell death in lymphoma and leukemia cell lines with rapid downregulation of p-AKT via the PDK-1 pathway. This apoptosis occurs in vitro using concentrations well below those that have been achieved in current clinical trials using these agents. Additional studies are being carried out to further delineate the mechanism of synergy as well as to further explore the impact of sequence of administration using this combination. Further studies are also planned to xplore the impact of the combination on primary human leukemia and lymphoma cells from the blood and bone marrow.
The Small Molecule Pan-Bcl-2 Inhibitor GX15-070 Induces Apoptosis In Vitro in Mantle Cell Lymphoma (MCL) Cells and Exhibits a Synergistic Effect in Combination with the Proteasome Inhibitor Bortezomib.
Authors: Galan Patricia Perez , Roue Gael , Villamor Neus , Campo Elias , Colomer Dolors 
Published: April 2020

Abstract
Mantle cell lymphoma (MCL) is a lymphoid malignancy derived from a subset of mature B-cells with coexpression of CD5 and the chromosomal translocation t(11;14)(q13;q32). MCL patients present an aggressive clinical course and poor response to conventional chemotherapy due to either rapid relapse after an initial response or primary resistance to drugs. Thus, there is currently a strong effort to develop compounds that target novel biological pathways. In addition, MCL cells overexpress the antiapoptotic proteins Bcl-2, Bcl-XL and Mcl-1. The dysregulated Bcl-2 pathway represents an important target for a new-mechanism therapeutic approach. In this context, the pan-Bcl-2 inhibitor GX15-070 that belongs to a new family of compounds which mimic BH3-only proteins by binding to multiple antiapoptotic Bcl-2 members and consequently eliciting Bax and Bak dependent cytotoxicity. GX15-070 is currently being evaluated in Phase I clinical trials. GX15-070 induced apoptosis in vitro in primary cells from MCL patients as well in MCL cells lines, at doses ranging from 0,5μM to 5μM. GX15-070 induced phosphatidylserine exposure, mitochondrial depolarization, ROS generation, Bax and Bak conformational changes and caspase activation. As expected, pan-caspase inhibitors did not block apoptosis in accordance with the mitochondria-targeted nature of this new compound. The susceptibility of MCL cells correlates with the expression of Bcl-2, Mcl-1 and Bcl-XL, both at protein and mRNA levels. Although GX15-070 exhibited significant in vitro cytotoxicity in MCL as a single agent, we tested also this compound in combination with proteasome inhibitors. Bortezomib (PS-341, Velcade®) has shown promising results in MCL treatment. One of the drawbacks of proteasome inhibitors is the undesirable accumulation, due to absence of degradation, of antiapoptotic factors which could impede or slow apoptosis, such as Mcl-1 protein. With this rationale we evaluated a combination of bortezomib with the pan-Bcl-2 inhibitor GX15-070 in vitro in MCL cells. Preincubation of MCL cells with GX15-070 at doses ranging from 0,1μM to 1μM before bortezomib (1–10nM) addition induced a synergistic cytotoxic effect (Chou Talalay CI =[0,26–0,575]). In vitro, this new combination allows to reduce bortezomib dose 5 to 20 times, depending on the individuals. Western blot analysis revealed that this combination reduced Mcl-1 levels and displaced Bak from Mcl-1 sequestration. Moreover, upregulation of the proapoptotic BH3-only protein Noxa by bortezomib, was further increased when combined with GX15-070. All these findings suggest that GX15-070 and bortezomib combination could be a new therapeutic approach for MCL treatment.
Dose Escalated CHOP + Etoposide Followed by Repetitive Autologous Stem Cell Transplantation (MegaCHOEP) with or without Rituximab for Primary Treatment of Aggressive NHL.
Authors: Glass Bertram , Kloess Marita , Reiser Marcel , Metzner Bernd , Truemper Lorenz H. , Loeffler Markus , Pfreundschuh Michael , Schmitz Norbert 
Published: April 2020

Abstract
Whilst safety and efficacy of adding rituximab to standard therapy has been reported, less information is available on the combination of rituximab (R) with HDT and autologous SCT. The German Study Group for High-Grade NHL (DSHNHL) developed a repetitive HDT program consisting of 4 courses dose escalated CHOP + Etoposide +/− R. Patients aged 18 to 60 years and elevated LDH at diagnosis were included. Mega-CHOEP consisted of four cycles of chemotherapy. Cycles 2 to 4 were followed by autologous SCT. At dose level 3 (DL3) cumulative doses of drugs were as follows: C 19.5 g/m2, H 280 mg/m2, O 8 mg, E 5.04 g/m2, P 2000 mg. When feasibility and safety of MegaCHOEP at DL3 had been demonstrated, rituximab was added (DL3R). Six cycles of rituximab (375 mg/m2) were given: one dose of rituximab prior to each cycle of chemotherapy and two additional doses 3 and 6 weeks after the last cycle of chemotherapy, respectively. From September 1999 to may 2001 35 pts with validated diagnosis of aggressive B-NHL received MegaCHOEP DL3, from June 2001 to February 2003, 72 pts received DL3R. There was a higher percentage of B-cell NHL patients with ECOG performance status &gt; 1 at DL3 (42.9 %) than at DL3R (19.7 %, p = 0.012) and as consequence more patients with age-adjusted IPI high risk at DL3 (34.3 %) than at DL3R (12.7 %, p = 0.029). We observed significant differences in the incidence of WHO grade III/IV infections (DL3 9 of 123 cycles (7.3%), DL3R 48 of 236 cycles (20.3%); p=0.001) and in the percentage of patients receiving the complete program of chemotherapy (DL3:91 %, DL3R 71%; p=0.017). There were no relevant differences in other types of non-hematologic toxicity or hematopoietic recovery between treatment groups. Median observation times of patients at DL3 and DL3R were 2.6 years and 4.5 years, respectively. In univariate analysis, there was a trend to better overall survival with rituximab (75%) than without rituximab (57%) at three years (p=0.168) and a significantly better freedom from treatment failure in patients receiving DL3R as compared to DL3 with 70 % vs 50 % at 3 years (p = 0.040). In a Cox regression model adjusted for IPI, there was a trend to a higher relative risk of treatment failure of MegaCHOEP without rituximab (RR 1.8); this, however, was not statistically significant (p = 0.087). Rituximab may have beneficial influence on lymphoma control in this setting; its use, however, is accompanied by higher incidence of serious infections which subsequently compromise the dose intensity of this aggressive treatment program. The DSHNHL is currently performing a randomized study comparing MegaCHOEP+R with CHOEP−14+R.
Rituximab in Relapsed Lymphocyte Predominant Hodgkin’s Disease (LPHD). Long-Term Results of a Phase-II Study from the German Hodgkin Lymphoma Study Group (GHSG).
Authors: Schulz Holger , Trelle Sven , Reiser Marcel , Sieber Marcus , Diehl Volker , Engert Andreas 
Published: April 2020

Abstract
Introduction: LPHD is a rare disease which accounts for 3–8% of all Hodgkin cases. Patients with LPHD relapse frequently and freedom from treatment failure is not significantly improved by intensification of polychemotherapy or radiotherapy. The malignant cells of LPHD are CD20+ and therefore the anti-CD20 antibody rituximab (R) may have activity with fewer adverse late effects.
Methods: This phase-II trial was initiated by the German Hodgkin Lymphoma Study Group (GHSG) to evaluate rituximab in patients with LPHD at first or higher relapse or progressive disease after at least one standard treatment. Histological slides were reviewed by a reference panel. Pts received 375mg/m2 of the anti-CD20 antibody rituximab once weekly for four weeks given as intravenous infusion in saline solution.
Results: Between 1999 and 2004 we treated twenty-one pts. with CD20-positive Hodgkin’s lymphoma according to the study protocol. Fourteen patients had stage I/II disease at the time of study entry and all patients were in their first to third relapse (median 2). The initial diagnosis of LPHD was confirmed in 17/21 cases. The remaining cases were reclassified as HD transformed to T-cell rich B-cell lymphoma (2) or CD20-positive classical HD (2). The overall response rate was 90%. Time to progression was 31 months (ms). The median follow-up 58 ms. Both T-cell rich B-cell lymphoma are in continous remission (PR 51ms+, CR 61ms+).
Conclusion: Single-agent therapy with rituximab is safe and showed high efficacy in relapsed LPHD. Therefore rituximab might be a non-toxic and efficient alternative treatment strategy compared to intensified chemo-and/or radiotherapeutic protocols in this young group of patients.
Adoptive Immunotherapy with Autologous CD25-Depleted and CD3/CD28-Costimulated T-Cells (ACTC) Enhances Numeric and Functional Lymphocyte Recovery after Cyclophosphamide - Fludarabine Chemotherapy in Patients with Low-Grade Follicular Lymphoma.
Authors: Andreadis Charalambos , Levine Bruce L. , Cotte Julio , Zheng Zhaohui , Mair Rebecca , Cannon Andrea , Harrow Jason , Traum Marcy , Chong Elise A. , Downs Lisa H. , Vogl Dan T. , Svoboda Jakub , June Carl H. , Schuster Stephen J. 
Published: April 2020

Abstract
Fludarabine-based chemotherapy combinations are highly effective in patients (pts) with low-grade follicular lymphoma (FL), but cause severe and long-lasting immunosuppression due to depletion of normal CD4 T-cells. Aside from increasing the risk of serious infections, this toxicity may limit the ability of the immune system to eliminate minimal residual disease. Adoptive immunotherapy using autologous CD25-depleted, CD3/CD28-costimulated T-cells expanded ex vivo (ACTC) may enhance immune reconstitution and improve disease control. We initiated a phase I study in pts with purine analog-naive relapsed/refractory FL (grades 1 and 2). After leukapheresis, pts receive 4 cycles of fludarabine (25 mg/m2) days 1–3 and cyclophosphamide (250 mg/m2) days 1–3. Four weeks after last chemotherapy, responding patients (CR, CRu, PR) receive escalating doses of ACTC prepared ex vivo from autologous T-cells collected prior to chemotherapy and depleted of regulatory CD4+/CD25+ cells, then expanded and activated using anti-CD3 and anti-CD28. Eight pts have been enrolled to date. Median age is 40.5 y (range: 32–64). Median number of prior therapies is 2 (range: 1–3). Two pts were withdrawn from the study due to hematologic toxicity related to the chemotherapy; one patient has not completed chemotherapy. Of the 5 pts completing chemotherapy, 3 pts achieved a CR and 2 pts achieved a PR; 4 pts received 5 x 109 and 1 patient received 1 x 1010 CD3+ ACTC. There have been no adverse events related to T-cell infusions. Median follow-up after ACTC infusion is 9 months (range: 2–15 months). Median time to CD4 count &gt;200 /uL was 29 days following T-cell infusion (range 28–127 days). CD4 counts increased in all patients by 1 month after T-cell infusion, with a median increase of 126% from baseline (range 50 to 484%). CD8 counts also increased, with a median increase of 82% (range −4 to 976%). All 5 pts receiving ACTC were anergic to candida antigen by Delayed Type Hypersensitivity (DTH) skin testing before chemotherapy. Three pts developed a positive DTH response to candida antigen 60 days after ACTC infusion. From the start of therapy for patients receiving T-cells, median follow-up is 14 months (range 7–20 months) with median progression-free survival not reached; 4 pts remain in remission and 1 patient had progression of disease. ACTC results in significant CD4+ lymphocyte recovery in previously treated pts receiving cyclophosphamide-fludarabine chemotherapy and compares very favorably with historical controls. T-cell dose escalation is ongoing.