bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunotherapy with Cytokine Induced Killer Cells in Solid and Hematopoietic Tumors: Preliminary Data.
Authors: Olioso Paola , Giancola Raffaella , Di Riti Maria , Accorsi Patrizia , Spadano Antonio , Natale Dino , Rossetti Roberto , Di Bartolomeo Paolo , Iacone Antonio 
Published: April 2020

Abstract
CIK cells are a unique population of CD3+CD56+ immune effector cells capable of lysing a broad variety of tumor cells targeted through a perforin based mechanism mediated by NKG2D.
We started a pilot clinical trial in patients with refractory lymphoma and solid tumors according to GMP guidelines that is currently ongoing.
Methods: CIK cells were generated from PBMNC and incubated in LifeCell culture bags in the presence of IFN-γ followed by IL-1β, OKT3 and IL-2. Expansion was assessed between day 21 and 28 and flow cytometric analysis was performed every week. Patients were monitored before and after treatment. Unpaired t-test was used to analyze for statistical significance. A p-value &lt;0.05 was considered statistically significant.
Results: At present 10 patients are enrolled: 6 advanced lymphomas, 3 metastatic kidney carcinoma and 1 hepatocellular carcinoma. The median number of transferred cells per patient was 19 x109 (6–37 x109) and the absolute number of CD3+CD56+ cells infused ranged from 1 to 16 x109 (median value 5 x107/Kg). Patients affected by solid tumors received in association low doses of rhIL-2 or α-interferon. Protocol adherence was excellent and the toxicity profile was favourable. Only 2 patients developed low-grade fever during the first cycle of infusion (5%) but promptly resolved without antibiotic treatment. After CIK cell infusion, in patient’s peripheral blood the absolute median count of PBLs, CD3+, CD8+ and CD3+CD56+ cells significantly increased with a p-value of 0.034, 0.025, 0.034 and 0.038, respectively. Clinical outcome appeared promising: 2 patients had complete response (1 metastatic kidney carcinoma and 1 hepatocellular carcinoma) and 2 patients had stabilization of disease (1 metastatic kidney carcinoma and 1 NHL) with a median follow-up of 12 months (range 4–14).
Conclusions: These preliminary data showed that adoptive immunotherapy with CIK cells is a safe therapy with some suggestion of efficacy that significantly enhances immune functions increasing absolute numbers of effector cells without side effects. If confirmed in larger scale studies, these promising results may have a favourable impact on conventional treatment strategy of malignancies.
A Better Strategy To Extend the Proliferative Lifespan of Human T-Cells In Vitro.
Authors: Roeth Alexander , Schneider Lisa , Baerlocher Gabriela , Duehrsen Ulrich 
Published: April 2020

Abstract
Telomerase maintains telomere length by adding TTAGGG repeats to the 3′-ends of chromosomes. In human T-cells telomerase is transiently expressed upon activation and stimulation and, as shown previously, telomerase levels are able to control their lifespan. To understand the effects of culture parameters on telomerase activity and cell lifespan is of critical importance to improve T-cell expansion. Here, we investigated the influence of culture conditions and of stimulation on lifespan, clonogenicity (number of positive wells), cell cycle and telomerase activity in T-cells in vitro. Isolated peripheral blood mononuclear cells (PBMCs) from healthy individuals were cultured in RPMI 1640 medium containing IL-2, different sera (10%) (fetal calf serum (FCS), AB serum and autologous serum) and stimulated with phytohaemagglutinin (PHA) and irradiated allogeneic mononuclear feeder cells or anti-CD3/CD28 beads. T-cells were counted and passaged weekly until senescence. Expanded T-cells were analyzed for cell cycle by DNA staining with propidium iodide (PI) at various time points after stimulation and telomerase activity was measured using the telomerase PCR ELISA. Clonogenicity was determined by limiting dilution using different culture conditions. The proliferative lifespan of T-cells expanded with PHA/feeder cells and autologous serum from two different donors was significantly increased compared to T-cells stimulated with PHA/feeder cells and AB serum (44.7 vs. 14.2 PDs; 42.4 vs. 27.4 PDs). In contrast, no or only little difference was found for T-cells expanded with anti-CD3/CD28 beads and autologous or AB serum (8.7 vs. 8.1 PDs; 15.5 vs. 9.6 PDs). Most likely the latter findings reflect the loss of CD28 expression on T-cells. Furthermore, the use of autologous serum also increased the clonogenicity to about 4-fold compared to the use of AB serum or FCS without any signs for differences in the fractions of cycling cells. Interestingly, T-cells cultured with autologous serum exhibited a higher telomerase activity at day 3 after stimulation and a reduced decline of telomerase activity at day 6 and 9 compared to cultures with AB serum. Our results demonstrate that the use of autologous serum combined with PHA stimulation and feeder cells remarkably extends the proliferative lifespan and clonogenicity of human T-cells in vitro. In addition, T-cells cultured in medium containing autologous serum exhibit higher and prolonged telomerase activity compared to cultures with AB serum. Further studies are ongoing to elucidate the underlying factors involved in those observations. Nevertheless, we suggest that autologous serum together with PHA stimulation and feeder cells is superior to expand human T-cells especially for clinical applications where large numbers of specific T-cells are required for adoptive transfer strategies in cancer patients with for example severe viral infections.
B-Aggressive Lymphoma (BAL) Family Proteins Have Unique Domains Which Modulate Transcription and Exhibit PARP Activity.
Authors: Aguiar Ricardo C.T. , Takeyama Kunihiko , He Chunyan , Kreinbrink Katherine , Shipp Margaret A. 
Published: April 2020

Abstract
BAL1 was originally identified in a genome-wide search for risk-related genes in diffuse large B-cell lymphoma (DLBCL). In pilot and subsequent independent series of primary DLBCLs, BAL1 expression was significantly higher in chemoresistant tumors. BAL1 encodes a nuclear protein with a duplicated N-terminal domain homologous to the non-histone region of histone macroH2A. Several lines of evidence suggest that macro domains may modulate transcription, including: high concentration of histone macroH2A in the inactive X chromosome, direct interference with transcription factor binding in a positioned nucleosome and structural similarity to DNA binding domains. In addition to N-terminal macro domains, the BAL1 protein includes a C-terminal region with homology to the catalytic domain of poly(ADP-ribose)polymerase (PARP) proteins. PARPs catalyze the transfer of ADP-ribose onto acceptor proteins, using NAD+ as a substrate. Poly(ADP-ribosyl)ation is a critical post-translational modification that regulates chromatin configuration, transcription and DNA repair. After characterizing BAL1, we recently identified two additional BAL family members, BAL2 and BAL3, with multiple N-terminal macro domains and putative C-terminal PARP catalytic domains. Given the emerging evidence that PARP family proteins represent possible cancer treatment targets and the reported association between BAL1 expression and chemoresistance in DLBCL, we performed detailed structure/function analyses of BAL family members. We demonstrate that BAL macro domains repress transcription when tethered via a Gal4 DNA binding domain to a TK promoter in classic transcription repression assays. In addition, we show that BAL2 and BAL3, but not BAL1, exhibit PARP activity. In agreement with these data, BAL1 lacks several critical donor and acceptor residues that are conserved in the BAL2 and 3 PARP active sites. Of interest, BAL family members with inactive or functional PARP domains differed in their ability to repress transcription. BAL family members are the only described proteins with both PARP and macro domains, underscoring the potential functional significance of this unique combination.
Active Combination Therapy of Bortezomib (Velcade) and Ibritumomab Tiuxetan (Zevalin) in an In Vivo Diffuse Large B-Cell Lymphoma Model.
Authors: Pervan Milena , Calimlim Jeffrey , Matso Dan , Said Jonathan W. , McBride William H. , de Vos Sven 
Published: April 2020

Abstract
Chemotherapy-resistant diffuse large B-cell lymphomas (DLBCL) pose an unresolved clinical problem and most patients die of their disease within months. The goal of this study was to test a novel, non-chemotherapy containing regimen for Non Hodgkin’s lymphomas, including DLBCL, by combining the proteasome inhibitor bortezomib (PS-341, VELCADE®) with radio-immunotherapy using ibritumomab tiuxetan (Zevalin™). Previously, we have shown that the combination of bortezomib and rituximab is an active and at least additive regimen in an in vitro and in vivo DLBCL model. These data supported an ongoing clinical trial using this combination in patients with low grade NHL. In addition, Pervan et al. identified the proteasome as a novel, sensitive target for ionizing radiation (Mol Cancer Res. 2005 (7):381–90), providing further rationale for this combination study. The combination treatment was evaluated in vivo in a SCID diffuse large B-cell lymphoma model. Bortezomib treatment did not alter the CD20 expression levels of SUDHL-16 cells in vitro. MMT assays showed an additive effect of bortezomib and gamma radiation on SUDHL-16 cell proliferation. In preliminary experiments, we identified the maximal tolerated dose of ibritumomab tiuxetan in our mouse model as 50 μCi/mouse. 2x106 SUDHL-16 cells injected s.q. in the flank resulted in local tumor growth within 4 weeks. When tumors became palpable, animals were stratified in 4 different treatment groups of 12 mice each. Mice received i.v. injections of either diluent, single agent bortezomib, single agent ibritumomab tiuxetan, or the combination of both. Ibritumomab tiuxetan was injected once i.v. at a dose of 20 μCi /mouse. Bortezomib was given i.v. as single dose of 0.01 mg/mouse. The combined treatment group received i.v. bortezomib as a radio-sensitizer 3 hrs prior to the injection of ibritumomab tiuxetan. All treatment schedules delayed tumor growth compared with controls. After 16 days of treatment, bortezomib reduced tumor progression by 52%, ibritumomab tiuxetan by 69%, and the combination treatment by 91%. Selected mice were sacrificed at day two and their tumors studied for potential immediate therapeutic effects. The Y90 content of tumors and normal tissues was measured and demonstrated targeting of Y90 to the xenograft. The tumors of the control and treatment groups did not differ by microscopic morphological appearance. In situ Tunel assays did not reveal increased apoptosis at treatment day two. In summary, bortezomib as a radio-sensitizer in combination with ibritumomab tiuxetan is an active and at least additive regimen in an in vivo DLBCL model.
Inhibition of Protein Kinase C beta by Enzastaurin (LY 317615) Induces Alterations of Key Regulators of Cell Cycle and Apoptosis in Mantle Cell Lymphoma and Synergizes with Chemotherapeutic Agents in a Sequence Dependent Manner.
Authors: Rieken Malte , Weigert Oliver , Pastore Alessandro , Hutter Grit , Zimmermann Yvonne , Weinkauf Marc , Hiddemann Wolfgang , Dreyling Martin 
Published: April 2020

Abstract
Mantle Cell Lymphoma (MCL) is a subtype of malignant lymphoma, characterized by a poor clinical outcome, a median survival time of three years and almost no long-term survivors. In previous studies it could be shown that constitutive overexpression of NFκB pathway plays an important role in the pathogenesis of MCL. The protein kinase C (PKC) family is essential in cell signal regulation effecting cellular growth, proliferation and apoptosis. Previous studies in diffuse large B-cell lymphoma suggest that PKCβ overexpression enhances B-cell proliferation and is associated with poor clinical prognosis. The pivotal role of PKCβ in activation of NFκB renders it a promising therapeutic target in the therapy of MCL. Five MCL cell lines (GRANTA 519, HBL-2, Jeko-1, NCEB-1, Rec-1) and two haematological control cell lines (Jurkat, Karpas 422) were treated with Enzastaurin in a proliferation inhibiting dose of 10μM, defined in initial pilot experiments. Real-time PT-PCR and protein expression (Western blot) levels of various regulators of cell cycle and apoptosis were determined at various time points of enzastaurin treatment (0.5 to 12 hours). In addition analysis of cell cycle and apoptosis were performed by flow-cytometry. All, cell lines were also exposed to different doses of enzastaurin in combination with cytarabine, fludarabine or mitoxantrone and analysed for efficacy by evaluating the combination index. Inhibition of proliferation was achieved in all cell lines in a dose and time-dependent manner, however susceptibility varied strongly within the cell lines characterized by IC 50 (24 hours) ranging from 8,0 μM in Jeko-1 to &gt; 50μM in GRANTA 519, Rec-1 and Karpas 422. Induction of apoptosis was detected in all cell lines according to susceptibility, and cell cycle analysis exhibits a moderate accumulation of and cells in the G2/M phase. Significant alterations of RNA expression was detected for BCL-2, cyclin D1 and p14 applying the Kruskal-Wallis-Test. Preliminary Western blot data confirm the altered levels of protein expressions. Synergistic effects of Enzastaurin combinations were sequence dependent, and resulted in a super-additive effect when the exposure of cells to cytostatic agents was followed by Enzastaurin treatment. No synergy effect was observed in combination with fludarabine. Our study revealed alteration of gene expression, critical for the regulation of cell cycle and apoptosis, after Enzastaurin treatment. Even more important, combination with specific cytostatic drugs demonstrated a synergistic efficacy in vitro. Thus, our data suggest that inhibition of PKCβ can overcome chemotherapy resistance by cellular survival mechanisms supporting the evaluation of such combinations in clinical trials.
Antileukemic Activity of Shepherdin, a Novel Targeted Inhibitor of the Survivin-Hsp90 Complex.
Authors: Gyurkocza Boglarka , Plescia Janet , Toke Madhavi , Greiner Dale , Colombo Giorgio , Lowry Philip A. , Altieri Dario C. 
Published: April 2020

Abstract
Cancer therapy now aims at disabling critical signaling pathways essential for tumor cell maintenance without affecting normal cellular functions. The molecular interaction between heat shock protein 90 (Hsp90) and survivin functions as a crossroad to preserve cell viability and cell proliferation in malignant cells, and offers tangible therapeutic prospects. We recently used structure-based rational design to identify Shepherdin, a cell permeable peptidomimetic from the survivin sequence Lys79-Leu87, which inhibits the complex between survivin and Hsp90 in tumor cells (Kd~80 nM). Here, we tested the activity of Shepherdin against a panel of acute leukemia cell lines (AML). We identified a shorter cell-permeable Shepherdin variant Lys79-Gly83 that docked in the Hsp90 ATP pocket by molecular dynamics simulations, bound recombinant Hsp90 in vitro, and competitively inhibited the survivin-Hsp90 complex. When administered in cell culture, Shepherdin rapidly (&lt;30 min) accumulated intracellularly in all AML cells tested, as well as in quiescent or phytohemagglutinin (PHA)-stimulated normal mononuclear cells. A brief, 30-min exposure of AML cells to Shepherdin was sufficient to induce complete cell killing by apoptotic and non-apoptotic mechanisms, with associated mitochondrial dysfunction and effector caspase activity. Conversely, Shepherdin did not affect colony formation of isolated CD34+ hematopoietic progenitors or viability of resting or proliferating normal mononuclear cells. When compared with known Hsp90 antagonists currently in the clinic, i.e. 17-AAG, Shepherdin was superior for rapidity and efficacy of antileukemic activity. Shepherdin induced collapse of Hsp90 chaperone functions in solid tumor cell types with destabilization of multiple Hsp90 client proteins, but did not affect the levels of pro-survival Hsp70 in AML cells, which is typically upregulated following Hsp90 inhibition by 17-AAG. In conclusion, Shepherdin provides a novel approach for rational therapy of acute leukemia.
The Novel Triterpenoid 2-cyano-3,12-dioxooleana-1,9-dien-28-oic Acid (CDDO) Induces Apoptosis of Human Diffuse Large B-Cell Lymphoma Cells (DLCL) through a Peroxisome Proliferator-Activated Receptor γ (PPARγ) Independent Pathway Which Is Enhanced by NFκB Inhibition.
Authors: Ray Denise , Morse Kimberly , Hilchey Shannon , Garcia Tatiana , Felgar Raymond , Maggirwar Sanjay , Phipps Richard , Bernstein Steven H. 
Published: April 2020

Abstract
Ligands for the transcription factor PPARγ are emerging as a new class of anti-tumor agents. Herein we report that the synthetic triterpenoid CDDO, a PPARγ ligand that induces PPARγ transcriptional activity in human DLCL OCI-Ly-19 cells, also induces cell death in human DLCL of both germinal center (OCI-Ly19) and activated B-cell phenotype (OCI-Ly10), cells which express the PPARγ protein. This effect of CDDO appears to be independent of PPARγ stimulated pathways since the functional antagonist of PPARγ, GW9662, which completely inhibits CDDO induced PPARγ transcriptional activity was unable to prevent CDDO induced cell death. Similar findings were seen using the additional PPARγ antagonists T0070907 and BADGE. CDDO induces cell death by inhibiting cell proliferation and inducing apoptosis as shown by Annexin-V and propidium iodide staining. As we have previously shown that PPARγ ligands inhibit NF-κB activity in B lymphocytes (J. Immunol2005; 174(7): 4060–9), we next examined the effect of CDDO on NF-κB in DLCL cells. Surprisingly, exposure of Ly19 cells to CDDO resulted in a dose-, and time-dependent increase in the activity of both the p50 and p65 subunits of NF-κB as determined by ELISA, by direct visualization of the nuclear translocation of p65 using indirect immunofluorescence assays, and by EMSA. The nuclear translocation of both the p50 and p65 NF-κB subunits was also confirmed by performing immunoblot analyses using nuclear fractions of CDDO-treated Ly19 cells. NF-κB activation was also observed in Ly10 cells exposed to CDDO. Follow-up experiments revealed that the activation of NF-κB in Ly19 cells by CDDO was due to proteolysis of inhibitory IκBα molecules. To determine whether the CDDO-induced NF-κB activation was a pro- survival mechanism, Ly19 and Ly10 cells were pre-treated with the NF-κB inhibitors SN50, helenalin or BAY 11-7082 and then exposed to CDDO for 24 hrs. In all cases, the NF-κB inhibitors significantly enhanced CDDO induced cell death suggesting that NF-κB activation is an anti-apoptotic mechanism elicited to protect the cell against CDDO cytotoxicity. Collectively, these studies suggest that; (a) CDDO (which will shortly be entering clinical trials for patients with acute myeloid leukemia) as a single agent may have significant clinical activity in patients with DLCL and; (b) the combination of CDDO with pharmacological inhibitors of NF-κB would be a rationale combination of novel agents to test in the context of clinical trials for patients with DLCL.
Updated Results from a Phase II Study of Galiximab (Anti-CD80) in Combination with Rituximab for Relapsed or Refractory, Follicular NHL.
Authors: Friedberg Jonathan W. , Leonard John P. , Younes Anas , Fisher David C. , Gordon Leo I. , Moore Joseph O. , Czuczman Myron S. , Miller Thomas P. , Stiff Patrick J. , Cheson Bruce D. , Forero-Torres Andres , Finucane Deborah M. , Leigh Bryan R. , Molina Arturo 
Published: April 2020

Abstract
Galiximab is a PRIMATIZED®, anti-CD80, monoclonal antibody with human IgG1 constant regions and macaque variable regions. CD80 is an immune costimulatory molecule that is constitutively expressed on the surface of follicular lymphomas. Modest single-agent clinical activity was demonstrated in a Phase I study in relapsed or refractory, follicular NHL (ORR=11%). Preclinical data show that galiximab + rituximab may be more effective than either antibody alone. Here we report updated results from the Phase II part of a multicenter study (Study 114–21) evaluating galiximab + rituximab for relapsed or refractory, follicular NHL. Patients (pts) were administered galiximab (500 mg/m2 qwk x 4) concurrently with a standard course of rituximab (375 mg/m2 qwk x 4). Rituximab-refractory pts (no response or a response with TTP&lt;6 months) were excluded. Study objectives were to evaluate safety, PK, and efficacy. Sixty-four pts received treatment. The median follow up is 14.5 months. Mean age at study entry was 59 yrs. The majority of pts (88%) were Stage III/IV, and FLIPI risk groups were distributed as good (27%), intermediate (39%), or poor (34%). All pts had received at least 1 prior lymphoma therapy; 42% were rituximab naïve. Galiximab infusions were delivered over 1 hr and were well tolerated. No DLTs were reported. Sixty-one (95%) pts experienced study-related AEs; the most common were lymphopenia (44%), leucopenia (38%), fatigue (38%), neutropenia (23%), and chills (23%). IWRC was used to evaluate response. An ORR of 64% was demonstrated: 17% CR, 14% CRu, and 33% PR. The median PFS was 12.1 months. Univariate analysis showed no correlation between response and baseline characteristics, although there was a trend for fewer responses in pts with elevated LDH, poor FLIPI score, and Grade III follicular histology. Cmax and AUC values were dose-proportional, with a mean serum half-life of 13 to 24 days. These results were retrospectively compared with 3 historical Biogen Idec studies of follicular NHL pts treated with a standard course of rituximab monotherapy. Baseline characteristics were similar; however, there was a higher incidence of rituximab-naïve pts in the rituximab monotherapy group (77%) compared with galiximab + rituximab (42%). The toxicity profiles for the 2 groups were similar. The median PFS was longer in the galiximab + rituximab group (12.1 mo.) than in the historical rituximab monotherapy group (9.4 mo.). These results suggest that galiximab can be safely combined with a standard course of rituximab and produce promising response rates and PFS. The toxicity profile and PFS with this combination compared favorably with historical rituximab monotherapy studies. A Phase III, randomized study is planned.