bluuebunny/crossref_metadata_2025_split huggingface dataset:
In Vitro and In Vivo Targeting and Therapy of an Antibody-Drug Conjugate (IMMU-110) in B-Cell Malignancies.
Authors: Sapra Puja , Stein Rhona , Pickett Jennifer , Govindan Serengulam V. , Cardillo Thomas M. , Damoci Christopher , Sheerin Agatha , Hansen Hans J. , Horak Ivan D. , Griffiths Gary L. , Goldenberg David M. 
Published: October 2019

Abstract
IMMU-110 is a drug immunoconjugate comprised of doxorubicin (DOX) conjugated to the humanized anti-CD74 monoclonal antibody (mAb), hLL1, at a DOX:mAb (mol/mol) ratio of 8:1. CD74 is a rapidly internalizing type-II transmembrane chaperone molecule associated with HLA-DR, and has high expression on human non-Hodgkin’s lymphoma (NHL) and multiple myeloma (MM) clinical specimens and cell lines. Here, we investigated the in vitro and in vivo efficacy of IMMU-110 in xenograft models of human NHL (Raji, Daudi) and MM (MC/CAR). In vitro cell binding of IMMU-110 with the CD74-positive cells was significantly higher than that of a non-specific isotype-matched mAb-DOX conjugate (DOX conjugated to a mAb against epithelial glycoprotein-1; DOX-hRS7), and was similar to that of naked hLL1. Both IMMU-110 and naked hLL1 bound CD74 with subnanomolar affinity. The in vitro cytotoxicity of IMMU-110 was significantly higher than non-specific antibody-DOX conjugate, DOX-hRS7, and was similar to free DOX in MC/CAR, Raji or Daudi human Burkitt’s lymphoma cells. In CD74-negative cell lines, IMMU-110 was significantly less toxic than free DOX, having similar cytotoxicity to DOX-hRS7. In vivo, IMMU-110 displayed a pharmacokinetic and biodistribution profile almost identical to that of hLL1 mAb. Both hLL1 mAb and IMMU-110 had a biphasic clearance from the circulation; the α and β half-life (t1/2) of IMMU-110 were 4.6 h and 157.9 h, respectively, and those of hLL1 were 5.4 h and 151.5 h, respectively. In biodistribution studies, no significant difference was observed between IMMU-110 and naked hLL1 with regards to normal tissue uptake. Neither IMMU-110 nor naked hLL1 mAb had a significant association with any normal body tissue. In therapy experiments, a single i.v. protein dose of 350 μg IMMU-110, injected 5 days after implantation of MC/CAR cells in SCID mice, resulted in curing 70% of the animals. Similar cure rates were observed when treatment with IMMU-110 was given 10 days after transplantation of MC/CAR cells. In the Raji xenograft model, 100% of animals were cured with a single protein dose of 120 μg IMMU-110, injected 5 days after implantation of cells. In survival studies, the efficacy of IMMU-110 was significantly better than naked hLL1, the combination of naked hLL1 and free DOX, or of a non-specific antibody-DOX conjugate, DOX-hRS7. In a tolerability study in SCID mice, no toxic effect of IMMU-110 was observed even at the highest dose tested (2.5 mg /mouse). In conclusion, treatment of B-cell lymphoma and myeloma xenograft models with single injections of IMMU-110 resulted in high levels of response and long-term survivors. IMMU-110 is being further developed as a potential therapeutic for the treatment of CD74-positive tumors.
Antibodies to TRAIL Receptors R1 and R2 Induce Apoptosis in Non-Hodgkin’s Lymphoma Cells.
Authors: Smith Mitchell Reed , Jin Fang , Joshi Indira 
Published: October 2019

Abstract
Monoclonal antibodies and chemotherapy can be effective, but not curative, therapy for non-Hodgkin s lymphoma (NHL). Rituximab acts by several mechanisms, including directly signaling apoptosis of CD20+ cells via the mitochondrial pathway involving caspase 9. Chemotherapy also activates this apoptotic pathway. TRAIL-R1 and -R2 signaling induces apoptosis via a pathway that activates caspase 8. Thus, we investigated the effects of agonistic monoclonal antibodies to TRAIL-R1 and -R2 on NHL cell lines (DoHH2, WSU-FSCCL and FC-TxFL2, each t(14;18)+, EBV-). TRAIL-R1 (HGS-ETR1) and -R2 (HGS-ETR2) antibodies were from Human Genome Sciences (Rockville, MD). FC-TxFL2 expressed the highest cell surface levels of TRAIL-R1 (DR4, target of HGS-ETR1) and TRAIL-R2 (DR5, target of HGS-ETR2). WSU-FSCCL expressed lower, but significant, levels of each. DoHH2 expressed dim TRAIL-R1 and the lowest levels of TRAIL-R2 of the 3 cell lines. IC50 (MTT assay after 72 hr incubation with antibody) was 0.25mcg/ml for WSU-FSCCL and FC-TxFL2 with either HGS-ETR1 or HGS-ETR2, while DoHH2 was minimally inhibited by either antibody. Both HGS-ETR1 and HGS-ETR2 antibodies induced dose dependent increases in apoptosis (annexin V assay) of WSU-FSCCL and FC-TxFL-2, but not DoHH2. Caspase activation (flow cytometry using fluorescent substrates; Western analysis) was seen 4–6 hr after antibody addition. As expected, caspases 3 and 8 were activated by both antibodies in the two sensitive cell lines. Caspase 9, however, was also activated. Thus, TRAIL-R1 and TRAIL-R2 antibody binding inhibits growth, induces apoptosis and activates caspases 3, 8 and 9 in NHL cells expressing the targets. The induction of caspase 9 suggests cross-talk between the extrinsic and intrinsic pathways, in which activation of caspase 8 leads to cleavage and translocation of bid, which in turn leads to activation of caspase 9, and ultimately caspase 3. We have confirmed that bid cleavage does occur in these NHL cells after HGS-ETR1 and HGS-ETR2 binding. We have further shown that both the pan-caspase inhibitor ZVAD and a specific caspase 8 inhibitor block HGS-ETR1 and HGS-ETR2 induced apoptosis. As predicted, a specific inhibitor of caspase 9 only partially blocks apoptosis. This suggests that combination of the agonist HGS-ETR1 and HGS-ETR2 antibodies with agents that act via the caspase 9 pathway would be rational combinations to test for therapeutic potential in NHL.
Oral Melphalan, Cyclophosfamide and Prednisone (MCP) in 72 Newly Diagnosed Waldenström’s Macroglobulinemia Patients: Results and Costs Analysis.
Authors: Petrucci Maria Teresa , Annibali Ombretta , Levi Anna , Tirindelli Maria C. , Fossati Carolina , Gallucci Cristiano , Del Bianco Patrizia , Mandelli Franco , Avvisati Giuseppe 
Published: October 2019

Abstract
Treatment of Waldenström’s macroglobulinemia (WM) is based on the use of oral alkylating agents, even though recently, more expensive drugs have been proposed for its treatment.The aim of this study was to report the results obtained combining oral melphalan, cyclophosphamide and prednisone (MCP) in 72 WM patients, and to compare the results and related costs with those observed using more aggressive and expensive protocols. Between July 1973 and April 2002, we have observed 100 newly diagnosed WM, of whom 72 were symptomatic and therefore requiring a treatment. The antineoplastic treatment consisted of melphalan (6 mg/m2, days 1–7), cyclofosfamide (125 mg/m2, days 1–7) and prednisone (40 mg/m2, days 1–7) administered orally for a total of 12 courses. Thereafter, responding and stable patients were maintained with chlorambucil (3 mg/m2) and prednisone ( 6 mg/m2) until progression. Levels ≥1.5 g/dL of IgM immunoglobulin associated to 30% of lymphoplasmocytic cells in the bone marrow aspirate were required for diagnosing WM. Among these patients, 46 (63%) were men and 26 (47%) were women. Median age, IgM Immunoglobulin and Hb levels were at diagnosis: 61 years (range 42–87 years), 3.8 gr/dl ( range 1.5–6.5) and 12 gr/dl ( range 6.5 – 16) respectively.
Of these patients, we evaluated response rate, overall survival (OS), response duration (RD), freedom from progression (FFP), event free survival (EFS) duration, toxicities and costs/course in € of the drugs utilized.Of the 72 treated patients,71 (98.6%) were evaluable and 55 (77.5%) achieved a response, 7 (9.9%) showed a disease stabilization and 9 (12.6%) disease progression. After a median follow-up of 72 months (range 3–195 months), median OS, RD, FFP and EFS were: 66, 64, 55 and 47 months, respectively. No grade III or IV WHO toxicities were observed and toxicity was limited to transient nausea, vomiting and mild neutropenia. The cost/course for MCP was € 13, similar to that of standard protocols based on chlorambucil, and much less expensive compared to the costs of more aggressive protocols proposed for the treatment WM, ranging from € 74 to € 3260.90.In conclusion,the MCP is, as chlorambucil based protocols, a cost effective and safe option for treating patients with WM. Moreover, the obtained results do not appear inferior to those so far reported with more expensive, aggressive and toxic intravenous protocols.
Role of Anemia in Survival of Elderly Aggressive Non-Hodgkin’s Lymphoma Patients Following Chemotherapy.
Authors: Zinzani Pier Luigi , Tani Monica , Molinari Anna Lia , Visani Giuseppe , Gentilini Patrizia , Guardigni Luciano , Stefoni Vitorio , Alinari Lapo , Marchi Enrica , Gabriele Annalisa , Musuraca Gerardo , Fina Maria Paola , Baccarani Michele 
Published: October 2019

Abstract
Recent literature data suggest that baseline anemia is an important poor prognostic factor for non-Hodgkin’s lymphoma (NHL) outcome. We evaluated the prevalence, risk factors, and prognostic value of anemia following chemotherapy in elderly patients with aggressive NHL treated with conventional front-line chemotherapy. Patients and Methods. The retyrospective sample included 168 patients with diagnosis of aggressive NHL treated between March 1993 and February 2001 with VNCOP-B regimen in five italian institutions. The model included sex, age, hemoglobin level following chemotherapy, and response to chemotherapy. Tumor remission at the end of chemotherapy was associated to a highly significant reduction in risk (RR, 0.215; p=0.0001); advanced staging before chemotherapy independently predicted a poorer survival (II-IV vs. I-II; RR, 1.55; p=0.0023). The prognostic value of hemoglobin level at chemotherapy end, adjusted for all above factors, was statistically significant (RR for 1-g/dL increment, 0.76; p= 0.0041). Our preliminary results suggest the prognostic role of hemoglobin value after chemotherapy in terms of overall survival. This concept introduces the potential role of anemia correction in improving the survival rate in elderly aggressive NHL.
Acceptable Outcomes for Patients Receiving Allogeneic Hematopoietic Stem Cell Transplantation (AlloSCT) for Relapsed Aggressive Non-Hodgkin’s Lymphoma (NHL).
Authors: Doocey Richard T. , Song Kevin W. , Toze Cynthia L. , Connors Joseph M. , Nevill Thomas J. , Gascoyne Randy D. , Barnett Michael J. , Forrest Donna L. , Hogge Donna E. , Lavoie Julye C. , Nantel Stephen H. , Shepherd John D. , Sutherland Heather J. , Voss Nicholas , Smith Clayton A. 
Published: October 2019

Abstract
Background: AlloSCT for the treatment of aggressive histology NHL is appealing on the grounds of providing both a tumour free graft and a graft versus lymphoma mechanism for disease clearance. Currently only modest experience has been acquired concerning this treatment modality because of the fear of increased treatment related mortality (TRM).
Methods: Retrospective review of 47 consecutive patients with aggressive histology NHL who proceeded to AlloSCT between 1987–2003 in Vancouver. Patients with Burkitt’s lymphoma and lymphoblastic lymphoma were excluded.
Characteristics: WHO classification histology: 24, diffuse large B cell (including 5 T cell Rich B cell and 3 primary mediastinal); 16, transformed indolent to diffuse large B cell; 7, peripheral T cell unspecified. Gender M:F 27:20. Median age at AlloSCT 40 years (range 19–55 years). Disease status at AlloSCT: 33 (71%) chemosensitive relapse; 12 (25%) primary resistant /resistant relapse; 2 (4%) first complete remission. 39 (83%) had advanced stage disease at SCT. Stem cell source: 33 (71%) HLA-identical sibling bone marrow; 3 (6%) HLA-identical sibling peripheral blood; 8 (17%) unrelated donor bone marrow; 3 (6%) unrelated donor peripheral blood. Cyclophosphamide /TBI conditioning was used in 81% of patients.
Results: 22 patients remain alive post AlloSCT with a median follow-up of 40 months (3–135 months). The 5 year overall survival and event free survival (EFS) for the 47 patients was 47% (95% CI 32% – 63%) and 43% (95% CI 27% – 59%) respectively. The 5 year EFS for the individual disease groups was diffuse large B Cell 37%; transformed indolent B Cell 12%; peripheral T cell - unspecified 86%. Chemosensitivity at SCT was associated with a 5 year EFS of 41% (95% CI 23% – 60%) as compared to 37% (95% CI 8% – 67%) for those with chemorefractory disease (p = 0.61). The cumulative incidence of TRM at 3 months and relapse at 3 years were 17% (95% CI 8% – 29%) and 32% (95% CI 18% – 47%) respectively for the entire 47 patients. Cumulative incidence of relapse at 3 years was 54% for patients with chemoresistant disease versus 25% for those with chemosensitive disease (p=0.049). The incidence of acute and chronic graft versus host disease (GVHD) was 57% and 51% respectively.
Conclusion: AlloSCT for aggressive NHL results in an acceptable TRM and favourable long term EFS and OS for patients with advanced stage disease having relapsed after primary treatment. Patient with chemoresistant disease can have a durable remission post AlloSCT although they are at increased risk of relapse.
A Defined Donor Activating Natural Killer Cell Receptor Phenotype Protects Against Leukemic Relapse after HLA-Identical Hematopoietic Stem Cell Transplantation.
Authors: Demanet Christian J. , Verheyden Sonja J. , Schots Rik J. 
Published: October 2019

Abstract
Objective: Killer immunoglobulin-like receptors (KIRs) on Natural Killer (NK) cells recognize groups of HLA class I alleles. Failure to recognize the appropriate KIR ligand on a HLA class I-mismatched cell triggers NK cytotoxicity. Because KIR and HLA genes are located on different chromosomes matching for HLA does mostly not result in matched KIR genes in related HLA-identical hematopoietic stem cell transplantation (HSCT). Moreover, KIR genotypes are very diverse in the population and therefore we explored whether this KIR diversity could have any Graft-versus-Leukemia (GvL) effect in related, HLA-identical HSCT.
Methods: A total of 65 consecutive patients with leukemia (AML, CML, ALL) who received a first transplant at our institution, were included in the analysis. The median age of the patients was 34 years. The conditioning regimen included TBI + cytosine arabinoside + cyclophosphamide in the majority of the patients (85%). The source of stem cells was bone marrow (72%) or peripheral blood (28%). The NK KIR genotype (6 inhibiting and 5 activating KIRs) and molecular HLA compatibility (locus A, B, C, DRB1, DQB1, DPB1) were studied in all patients and their sibling donors.
Results and conclusion: Our analysis showed that the combined presence of the activating KIR receptors 2DS1 and 2DS2 on the donor NK cells is of major importance in protection against leukemic relapse in HLA-identical HSCT. In the patient group with a KIR2DS1(+)2DS2(+) donor, relapse was significantly lower (2/15; 13%) compared to all other donors (23/50; 46%) (P=0.01). Additionally, we found a direct relationship between the number of HLA class I - activating KIR interactions and relapse. The more activating KIR receptor-HLA-C ligand interactions possible, the less relapse observed (P=0.05). However, we cannot exclude at this moment that a hitherto unknown ligand (such as a viral protein or a tumor associated or -specific protein) could substitute for a HLA class I antigen. HLA class I down-regulation is frequently observed on leukemic cells and decreases the interaction with inhibitory NK receptors. The presence of sufficient donor activating NK receptors to overcome the remaining inhibition is likely the mechanism that mediates the elimination of residual leukemic cells.
Tumor Antigen Immunization of Sibling Stem Cell Transplant Donors in Multiple Myeloma.
Authors: Neelapu Sattva S. , Munshi Nikhil C. , Jagannath Sundar , Watson Thelma M. , Pennington Robin , Barlogie Bart , Kwak Larry W. 
Published: October 2019

Abstract
Strategies to enhance the specific anti-tumor effect of the graft without increasing the risk of graft-versus-host disease (GVHD) are needed to improve the outcome in allotransplant recipients. In this pilot study, we vaccinated sibling donors against a defined, patient tumor-derived antigen prior to bone marrow harvest. The objective of this strategy was to induce tumor antigen-specific immunity in healthy donors and to transfer this immunity to the recipients by bone marrow transplantation (BMT). For this purpose, the unique antigenic determinants (Idiotype [Id]) of the immunoglobulin secreted by the myeloma tumor can serve as a safe, tumor-specific antigen for active immunotherapy. The vaccine formulation consisted of Id isolated from the plasma of myeloma patients chemically conjugated to a carrier molecule keyhole limpet-hemocyanin (KLH) and administered together with GM-CSF as an adjuvant. Immunization of the HLA-matched sibling donors (n = 5) prior to bone marrow harvest, induced specific cellular and/or humoral immune responses against Id and KLH in all five donors. No short-term or long-term toxicities were observed in any of the donors following vaccination. The recipients received priming vaccination pretransplant and booster immunizations starting approximately 3 months after the myeloablative transplantation. Following BMT, two patients died within 30 days due to transplant related complications. Id-specific and KLH-specific T cell responses were detected in all three surviving patients post-, but not pre-BMT. In at least one patient, Id-specific T cell immunity persisted beyond 18 months after completion of the vaccination schedule. All three surviving patients converted from a partial response to a complete response following BMT. Two of the three patients (both recipients of immune BMT from syngeneic twins) remain in complete remission 8 years and 7 years after the transplant, and the third patient developed chronic steroid-dependent GVHD and died of renal failure after 5.5 years while in complete remission from the disease. Our results suggest that Id vaccination induces specific T cell immunity in the donors that is transferable by BMT, is associated with a prolonged disease-free survival in recipients and may represent a novel strategy to enhance the graft versus myeloma effect. These encouraging results have prompted us to initiate a clinical trial to evaluate this approach of donor immunization in the context of non-myeloablative allogeneic peripheral blood stem cell transplantation in myeloma patients.
CD40 Ligand Stimulation of Multiple Myeloma Cells Results in Upregulation of Surface Expressed Heat Shock Proteins and Increased Antigenicity.
Authors: Gullo Charles A. , Hwang William , Au Melvin , Greenfield Edward A. , Anderson Kenneth C. , Teoh Gerrard 
Published: October 2019

Abstract
Effective immune-based therapies against the plasma cell malignancy, multiple myeloma (MM), are currently lacking. Identification of novel antigens (Ag) on the surface of MM cells to use as cellular targets for the destruction of cancer cells by the body’s immune system has been of great interest. We and others have demonstrated that CD40 stimulation of MM cells results in marked upregulation of membrane bound proteins such as Ku86. Using CD40 triggered MM cells as immunogens and hybridoma technology; we generated a monoclonal antibody (mAb), 6D11, that recognizes a CD40 induced cell membrane Ag on MM cells. This Ag is detectable on the surface of MM cells using indirect immunofluorescence flow cytometric analysis. Moreover, in Western immunoblotting assays, 6D11 mAb reacts with a 94 kDa protein, which is strongly associated with a 78 kDa protein. Using high performance liquid chromatography and protein microsequencing, we confirm that these proteins are the heat shock proteins (HSP), glucose-regulated peptide 94 (GRP94) and GRP78, respectively. These data were confirmed using co-immunprecipitation experiments. Furthermore, we demonstrate through indirect immunofluorescence flow cytometric analysis and quantitative real time reverse transcription polymerase chain reaction (RQ-PCR) that CD40 ligand (CD40L) stimulation of MM cells results in rapid upregulation of both GRP94 and GRP78. Since HSPs have been shown to play a role in both Ag presentation, as well as the intracellular transport of cellular Ags, it is tempting to speculate that cell membrane expression of tumor-specific peptides could also be induced via CD40 triggering. Accordingly, CD40 induced cell membrane HSP expression resulted in increased antigenicity as determined by increased co-stimulatory molecule expression on Ag presenting cells (APC) and by increased immunoreactivity in mixed lymphocyte reactions (MLR). This suggests that CD40 induced HSP expression may indeed result in increased recognition of MM cancer by the immune system. Our study therefore supports the development of CD40-based targeted cell therapies against MM.
In Vitro Activity of a Novel Small Molecule Cyclin Dependent Kinase Inhibitor, CYC202 (seliciclib or R-Roscovitine), in Multiple Myeloma (MM).
Authors: Raje Noopur , Kumar Shaji , Hideshima Teru , Ishitsuka Kenji , Roccaro Aldo , Yasui Hiroshi , Hamasaki Makoto , Shiraishi Norihiko , Chauhan Dharminder , Munshi Nikhil C. , Green Simon R. , Anderson Kenneth C. 
Published: October 2019

Abstract
Perturbation of cell cycle is central to tumorigenesis, resulting in unrestricted cell proliferation. Cyclin dependent kinase (CDK) inhibitors have the potential to induce cell cycle arrest, followed by apoptosis, in cancer cells. CYC202 (seliciclib or R-roscovitine; Cyclacel, Dundee, UK) is a potent inhibitor of CDKs currently undergoing phase II clinical testing. In vitro data has shown this tri-substituted purine to have greatest potency against CDK2/cyclin E, although it also inhibits CDK2/cyclin A, CDK7/ cyclin H, and CDK9/cyclin T. It has been tested against a broad range of tumor cell lines, ultimately inducing cell cycle changes and apoptosis. This orally bio-available compound has also induced regression of human tumor xenografts in nude mice, prompting its clinical testing. Phase I studies have demonstrated favorable pharmacokinetics and toxicity profiles, and phase II trials are currently ongoing in combination with gemcitabine /cisplatin for non-small cell lung cancer and as a monotherapy in B-cell malignancies. We have previously shown that MM cell lines demonstrate relatively low levels of p21WAF1, suggesting that CDKs are constitutively activated, thereby promoting uncontrolled cell cycle regulation, growth, and proliferation. Conversely, inhibiting CDK activity may therefore trigger MM cell growth inhibition. Here we studied the in vitro activity of CYC202 in MM cells. Our data demonstrates that CYC202 has potent cytotoxic effects against MM cells that are both sensitive (MM1.S, RPMI 8226, U266, H929) and resistant (MM1.R, Dox-40, LR5, MR 20) to conventional chemotherapy. MM cell line cytotoxicity, as evidenced by MTT assays, is noted at 24 hours, with IC50 ranging from 25–50 mmol. In contrast, this dose was not toxic to normal peripheral blood mononuclear cells. Cell cycle analysis demonstrated an increase (35–50%) in MM cells in sub-G1 phase at 24 hrs induced by CYC202 (25 mmol), suggesting that CYC202 triggers apoptosis. Caspase-8 and poly ADP-ribose polymerase (PARP) cleavage, evidenced by western blot analysis further confirmed apoptosis of MM cells. Importantly, CYC202 triggered a rapid down-regulation of MCL-1, a known anti-apoptotic protein in MM. Treatment of MM cells with CYC202 also resulted in decreased phosphorylation of retinoblastoma protein. Protein expression of certain CDKs, specifically CDC2, CDK4, and CDK6, were also down-regulated after treatment. Ongoing studies are delineating the specific signaling cascades affected by CYC202 treatment. These studies provide the pre-clinical basis for a clinical trial of CYC202, either alone or in combination with other agents to improve patient outcome in MM.