bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Phase 1 Dose Escalation Study of a Fully Human, Antagonist Anti-CD40 Antibody, HCD122 (Formerly CHIR-12.12) in Patients with Relapsed and Refractory Multiple Myeloma.
Authors: Bensinger William , Jagannath Sundar , Becker Pamela S. , Anderson Kenneth C. , Stadtmauer Edward A. , Aukerman Lea , Fox Judith , Girish Sandhya , Bilic Sanela , Guzy Serge , Solinger Alan , Dort Sherri , Wang Yongyu , Hurst Deborah 
Published: October 2019

Abstract
HCD122 is a novel, fully human, IgG1 antagonistic monoclonal antibody targeting the CD40 receptor. This antibody blocks CD40-mediated signaling and is a potent mediator of antibody-dependent cellular cytotoxicity (ADCC). Previous preclinical investigation confirmed expression of CD40 on myeloma cells in the majority of patients and reported antitumor activity of HCD122 against multiple myeloma cells ex vivo (Tai, Y et al. Cancer Res2005; 65(13): 5898–5906). This ongoing phase 1 study will determine the maximum tolerated dose of CHIR-12.12 in multiple myeloma patients (pts) who are relapsed or refractory after at least one prior therapy. Planned dose levels are 1, 3 and 10 mg/kg administered IV once weekly for 4 weeks. Each dose group will enroll 3–6 pts to evaluate safety, pharmacokinetics (PK) and clinical response. To date, 9 pts have been treated at 2 dose levels: 3 pts at 1 mg/kg and 6 pts at 3 mg/kg. Median patient age is 65 yrs (46–81 yrs); median number of prior therapies is 3 (2–12). No dose limiting toxicity (DLT) occurred at the 1mg/kg dose level. At 3 mg/kg, 1 DLT of grade 4 thrombocytopenia occurred in 1 pt. No other grade 3 and 4 lab abnormalities and adverse events have been reported. In 7 pts with available data, infusions were well tolerated, with easily managed grade 1–2 toxicities, primarily chills (5 pts), nausea (3 pts), pyrexia (2 pts), and arthralgia (2 pts) mainly reported during the first infusion. Preliminary PK analysis showed more than dose proportional - increase in Cmax and AUC at the 3 mg/kg dose level compared to the 1 mg/kg dose level. At the 3 mg/kg dose, antibody accumulation occurred week-to-week; the mean Cmax after the fourth infusion on Day 22 was 126.1 mg/mL(range 52 – 195 ug/mL) and HCD122 levels were measurable up to Day 57 and in one patient up to Day 99. One week after the last 3 mg/kg dose, trough levels ranged from 28 to 109 mg/mL. Of the 3 pts at 1 mg/kg, one showed stable disease (SD) for >23 weeks and two had progressive disease (PD) by week 5. Of the 6 pts at 3 mg/kg, one had partial response (PR) at week 9 and was confirmed at week 15, one had SD for > 5 weeks, and 4 had PD at week 5. One pt with PD terminated the study before final safety evaluation, and must be replaced before assessment of the 3mg/kg dose level is complete. Thus, in preliminary studies, HCD122 appears to be safe and well tolerated to date at doses of 1 mg/kg and 3 mg/kg weekly for 4 doses and shows promising anti-myeloma activity. Enrollment is continuing to determine MTD.
Kit Signaling Regulates Mitf Expression in Mastocytosis.
Authors: Lee Youl-Nam , Noel Pierre , Shahlaee Amir , Carter Melody , Kapur Reuben , Wayne Alan , Metcalfe Dean D. , Takemoto Clifford 
Published: October 2019

Abstract
Mastocytosis is a heterogeneous disease arising from abnormal proliferation of mast cells. Activating mutations in codon D816 of the tyrosine kinase receptor, c-kit, are found in the majority of adult patients with systemic mastocytosis, an aggressive form of the disease. Constitutive activation of the Kit signaling pathway is critical to the transformed phenotype, and thus understanding how this pathway regulates downstream events is of great importance. A number of transcription factors are also essential to mast cell development, including the Microphthalmia-associated transcription factor (Mitf). We examined Mitf expression in bone marrow biopsies from nine patients with systemic mastocytosis by immunohistochemistry; we found that Mitf is highly expressed in all cases with the D816V mutation. In contrast, Mitf is not highly expressed in non-malignant mast cells in the bone marrow from patients with aplastic anemia and leukemia, suggesting thatMitf expression is regulated by Kit-dependent signalsMitf may play a role in the transformed phenotype of mastocytosis.We show that in normal mast cells, Kit signaling markedly upregulates Mitf expression. In both normal and malignant mast cells, pharmacologic inhibitors of Kit, and the downstream kinase, PI3K, block Mitf expression. To examine whether Mitf is required for transformed phenotype from constitutive Kit signaling in mast cells, we have used a shRNA-expressing lentivirus to knockdown Mitf expression in mastocytosis cell lines. We found that silencing of Mitf markedly impaired growth in proliferation and colony forming cell assays. This work demonstrates a link between two critical factors, Kit and Mitf, in the development of malignant mast cell disease.
Concomitant Myelofibrosis and Plasma Cell Disorders: The Mayo Clinic Experience.
Authors: Mesa Ruben A. , Rajkumar S. Vincent , Schwager Susan , McClure Rebecca , Powell Heather , Lasho Terra , Kyle Robert A. , Tefferi Ayalew 
Published: October 2019

Abstract
BACKGROUND: Bone marrow fibrosis might represent either a primary myeloid malignancy such as myelofibrosis with myeloid metaplasia (MMM) or a reactive phenomenon associated with other malignancies or inflammatory/infectious processes. The development of myelofibrosis in the context of a plasma cell dyscrasia is uncommon and incompletely understood from the standpoint of both pathogenesis and clinical implications. We sought to determine the origin and clinical ramifications of this phenomenon based on a consecutive series of patients with plasma cell disorders (PCD).
METHODS: A retrospective search of institutional databases was performed for clinical cases of PCD with the concomitant observation of myelofibrosis. Bone marrow aspirates and trephines from the time of diagnosis of both PCD and myelofibrosis were reviewed. Clinical history was abstracted for clinical manifestations as well as disease course. DNA from archived cytogenetic pellets, when available, from the time of diagnosis, was screened for the presence of JAK2V617F.
RESULTS: Among 7587 patients with a PCD seen over the last 30 years at our institution, 29 (0.3%) displayed concomitant myelofibrosis (median age 59 years, range 27–77; 52% males). The specific PCD was multiple myeloma in 25 patients, smoldering myeloma in 2, plasmacytoma in 1, and heavy chain disease in 1. Four patients (14%) had organomegaly and bone marrow histological features most consistent with MMM. Otherwise, myeloproliferative disorder (MPD)-characteristic features were infrequent and included leukocytosis and/or thrombocytosis in only 2 patients. None of the patients displayed thrombohemorrhagic complications. Archived DNA was available in 7 cases without either clinical or bone marrow histological evidence of a MPD and yet revealed the presence of JAK2V617F in 6 of the 7 cases (86%). In addition, one other patient without MPD phenotype displayed a monosomy 5 abnormality. To date, 2 patients, one with and one without a MPD phenotype underwent leukemic transformation. All other causes of death were related to PCD-associated complications. Median survival (Kaplan-Meier) for the patients with multiple myeloma was 50 months. Survival stratified by the myeloma international staging system (ISS) was similar or better than expected (published controls of 62, 44, and 29 months): median survivals for this series of 73, 51 and 37 months for stage I, II, and III disease, respectively.
CONCLUSIONS: Myelofibrosis that accompanies a PCD often represents an underlying MPD and does not negatively impact the natural history of the associated PCD.
The Jak2 V617F Oncogene Associated with Polycythemia Vera Requires a Functional FERM Domain for Transformation and for Expression of the Myc and Pim Proto-Oncogenes.
Authors: Sattler Martin , Crowley Brian J. , Gonneville Jeffrey R. , Ho Jeremy , Hudon Heidi H. , Podar Klaus , Royer Yohan , Constantinescu Stefan N. , Griffin James D. 
Published: October 2019

Abstract
The V617F activating point mutation in Jak2 has been shown to be associated with a proportion of patients with myeloproliferative disorders, including polycythemia vera, essential thrombocythemia, and idiopathic myelofibrosis. In normal hematopoietic cells, Jak2 signals only when associated with a growth factor receptor (such as the Epo receptor), but the role of receptors in transformation by Jak2V617F has been controversial. Constructs expressing wild type or Jak2V617F, with and without a point mutation in the FERM domain (Y114A) required for receptor binding, were introduced into BaF3 cells already expressing a human Epo receptor. These cells were then evaluated for growth factor independence, response to growth factors, and activation of several critical Jak2 signaling pathways. We found that, whereas BaF3-EpoR cells are readily transformed by Jak2V617F to Epo-independence, the addition of the FERM domain mutation blocked transformation to factor independence. Also, the FERM domain was required for induction of reactive oxygen species by Jak2V617F. Further, while cells expressing Jak2V617F had constitutive activation of STAT5, cells expressing Jak2 V617F/Y114A did not, suggesting that signaling is defective at a very proximal level. In addition, Jak2V617F induced expression of the Myc and Pim proto-oncogenes, which are known to cooperate in the transformation of hematopoietic cells. The expression of both proteins was dependent on a functional FERM domain. Finally, we evaluated the role of active STAT5 in transformation of BaF3 cells by introducing a STAT5 mutant that is constitutively activated, and found that activation of STAT5, by itself, was sufficient to induce Jak2V617F-dependent downstream targets believed to be critical for transformation, including Myc and Pim. Overall, our results suggest that constitutive activation of Jak2 requires an intact FERM domain for a transforming phenotype, and is necessary for activation of the major target of Jak2, STAT5. These results also suggest that small molecules that target Jak2, the FERM domain, or STAT5 would have activity in diseases associated with V617F mutations.
Responses to Thalidomide Combined with Prednisone Therapy for Myelofibrosis with Myeloid Metaplasia.
Authors: Qian Shenxian , Tan Junfeng , Shi Pengfei , Gao Gaqian 
Published: October 2019

Abstract
Myelofibrosis with myeloid metaplasia (MMM) is currently classified as a classic (i.e. not yet molecularly defined) myeloproliferative disorder (MPD), along with essential thrombocythemia (ET) and polycythemia vera (PV). All three MPDs represent stem-cell-derived clonal myeloproliferation that, in the case of MMM, is accompanied by an intense bone marrow stromal reaction that includes collagen fibrosis, osteosclerosis, and angiogenesis. To present the results of a long-term analysis of 2 sequential phase 2 trials of thalidomide (alone or in combination) for palliation of myelofibrosis with myeloid metaplasia (MMM). We analyzed (March 2003 to August 2005) initial and long-term outcomes from 33 patients with symptomatic MMM who had enrolled in either our thalidomide single-agent trial (n=12) or our trial of low-dose thalidomide (50 mg/d) combined with prednisone (n=21). Among the 33 study patients, 18 (54%) showed some improvement in their clinical course. Response rates for specific end points included improvements in anemia (12 of 33 [36%]), thrombocytopenia (8 of 12 [66%]), or splenomegaly (5 of 30 [17%]). The combination of low-dose thalidomide and prednisone, as opposed to single-agent thalidomide, was better tolerated and more efficacious. After a median follow-up of 17 months (range, 9–27 months), 10 of 33 patients (33%) showed an ongoing response, including 8 patients in whom protocol treatment has been discontinued for a median of 21 months (range, 16–27 months). Durable treatment responses were documented for only anemia and thrombocytopenia. Treatment response was not affected by the baseline status of bone marrow fibrosis, angiogenesis, osteosclerosis, cytogenetics. Unusual drug effects, all reversible, included leukocytosis (8 patients) and/or thrombocytosis (6 patients). Thalidomide (alone or combined with prednisone) is an effective first-line treatment of symptomatic anemia or thrombocytopenia in MMM. Thalidomide-based therapy has the potential to produce durable responses in MMM-associated cytopenias, even after discontinuation of the drug.
Cytogenetic Abnormalities in Essential Thrombocythemia: Prevalence and Prognostic Significance.
Authors: Wolanskyj Alexandra P. , Gangat Naseema , Schwager Susan M. , Ketterling Rhett P. , Tefferi Ayalew 
Published: October 2019

Abstract
Objectives: We conducted a study to describe the cytogenetic findings as well as clinical correlates and long-term prognostic relevance of abnormal cytogenetics at the time of diagnosis of Essential thrombocythemia (ET), in terms of clinical presentation, disease transformation into more aggressive myeloid disorders, and life expectancy.
Patients and Methods: The study cohort consisted of a consecutive group of patients with ET who fulfilled the World Health Organization (WHO) diagnostic criteria, in whom bone marrow biopsy was performed at diagnosis, with interpretable cytogenetic analysis obtained in all cases.
Results: A total of 403 patients were studied (median age, 56 years; median follow-up 64 months). The prevalence of abnormal cytogenetics at presentation was 6.7 % (27 of 403). The most common cytogenetic anomalies identified included trisomy 9 (4 patients), trisomy 8 (3 patients) and deletion 20q (3 patients). Parameters at diagnosis that were significantly associated with abnormal cytogenetics included palpable splenomegaly (p=0.03), current tobacco use (p=0.04); venous thrombosis (p= 0.019), extreme thrombocytosis (i.e., platelet count >1500 × 109/L, p = 0.03) and anemia with a hemoglobin of less than 10 g/dl (p=0.02); but did not include Jak2 mutation status, nor advanced age (≥ 60 years). During follow up, patients with abnormal cytogenetics were more likely to experience venous thrombosis (p=0.02) but not shorter survival [figure 1], transformation to AML, MDS or MMM, nor a greater requirement for cytotoxic therapy.
Conclusion: Cytogenetic anomalies at presentation are relatively uncommon in ET, and do not predict a greater predilection towards evolution into more aggressive myeloid disorders, nor inferior survival.
Fig. 1 Overall Survival According to Normal vs Abnormal Karyotype Fig. 1. Overall Survival According to Normal vs Abnormal Karyotype
Ex-Vivo Expansion and mRNA Transfection of Cord Blood Derived Natural Killer Cells with Preserved Cytotoxicity.
Authors: Tuncer Hande H. , Betancur Monica M. , Boissel Laurent H. , Friedman Robb S. , Klingemann Hans G. 
Published: October 2019

Abstract
Natural Killer (NK) cell-mediated cytotoxicity can control leukemia relapse while protecting patients from graft-versus-host disease (GVHD) after allogeneic stem cell transplant. Cord blood (CB) is rich in NK cells with similar properties of proliferation and cytotoxicity as adult blood NK cells. Hence these cells are attractive for developing strategies to eliminate residual disease after cord blood transplant. In this study, CB mononuclear cells were cryopreserved after CD3 depletion performed by immunomagnetic microbead selection (Miltenyi Biotec, Auburn, CA). After thawing, cells were plated for NK expansion, with or without a feeder layer of irradiated cord mesenchymal (UCM) cells either from the same (autologous) or from an unrelated (allogeneic) cord donor, with or without IL-2 (1000 IU/ml), IL-15 (10 ng/ml), IL-3 (10ng/ml) and Flt3 (10ng/ml). At a median of 19 days of culture (range 14–21), there was significantly higher expansion (range 3.5–72 fold) of CD56+/CD3− cells with the UCM feeder layer and cytokines compared to controls (mean 21.2 ± 20.8 fold increase vs 1.6 ± 0.9 fold increase with feeder layer only and 1.8 ± 0.89 fold increase with cytokines only, p=0.039 and p=0.041 respectively). There was no significant difference in NK expansion between autologous and allogeneic UCM feeder layers (29.6 ± 26.8 vs. 12.8 ± 8.9 fold, p=0.243). Expanded CB-NK cells were then tested for cytotoxicity against K562 cells using a calorimetric assay with PKH67-GL. CB-NK cells expanded either with autologous or allogeneic UCM feeder layers displayed enhanced cytotoxicity compared to controls plated with cytokines only (91.78±0.7% vs. 82.5±1.8%, p=0.003 and 89±2.3% vs. 83.7±0.18%, p=0.056, respectively). In order to test whether expanded CB-NK cells can be transfected for ultimately targeting malignant cells, we electroporated expanded CB-NK cells with mRNA produced from plasmid GFP-DNA by in vitro transcription. Flow cytometry was used to detect viability, which was 94%, 92% and 93% for non-transfected, GFP-DNA and GFP-mRNA samples respectively. GFP-mRNA expression at 24 hours was significantly higher (range 36.6–50.8%, mean 42.8±5.2%) compared GFP-cDNA controls (mean 4.2±0.35%, p<0.001). Mean GFP-mRNA expression was 35%, 31% and 16.5% at 48, 72 and 144 hours respectively. In summary, CB-NK cells can be effectively expanded with a feeder layer of UCM cells while preserving cytotoxicity and can also be genetically modified by mRNA transfection.
Alloreactivity of Natural Killer Cell Clones Against Childhood Precursor-B Lymphoblastic Leukemia Cells Is Determined by Differential KIR Expression.
Authors: Feuchtinger Tobias , Pfeiffer Matthias , Pfaffle Alexander , Telschik Heiko-Manuel , Handgretinger Rupert , Wernet Dorothee , Schumm Michael , Lotfi Ramin , Lang Peter 
Published: October 2019

Abstract
Relapse after allogeneic stem cell transplantation of patients with acute lymphoblastic leukemia (ALL) remains a major problem. A beneficial impact of alloreactive NK cells has been reported for myeloid malignancies, but has been questionable for B-lineage-ALL. Here we show that the majority of NK cell clones from healthy donors could effectively lyse primary blasts from a representative childhood precurser-B-ALL and demonstrate the relevance of differential surface expression of KIRs (killer cell immunglobulin like receptors CD158a, CD158b and CD158e) for antileukemic alloreactivity. More than 79% of clones exerted specific lysis >40% against primary ALL blasts. Mismatch of effector/target HLA-C type (KIR-ligand incompatibility) did not correlate with antileukemic alloreactivity, although non-malignant lymphoblastoid cell lines were lysed according to KIR-ligand incompatibility. In contrast differential surface expression of the three major KIRs showed significant impact on the antileukemic activity against precursor-B-ALL blasts. NK clones with none of the three KIRs or a single KIR that recognized no ligand, were not inhibited by the targets and exerted higher lysis (p = <0.03) in comparison to NK clones with expression of one or more KIRs with a ligand on the ALL blasts. In conclusion the differential surface expression of KIRs is relevant to predict NK cell activity against childhood lymphoid leukemia.
Functional Characterization of a Continuous Natural Killer Cell Line, KIL, In Vitro and in Adoptive Immunotherapy.
Authors: Tsai Schickwann , Leukel H. Christopher , Morita Chikako , Heikens Marc J. , Cao Thai M. 
Published: October 2019

Abstract
We have recently established a continuous natural killer (NK) cell line, KIL (for Killer Lymphocyte), from a coculture of normal murine whole bone marrow (WBM) and the OP-9 stromal cell line expressing the Notch ligand, Jagged2, in the presence of stem cell factor and IL-7 (Dehart et al. Blood2005;105:3521). A clonal prototype, KIL C.2, was derived from a C57BL/6 mouse and has the CD3−NK1.1+CD122+ phenotype, expresses the NKG2D activating receptor, and proliferates extensively upon in vitro stimulation with IL-2 (20ng/ml). We now show that KIL C.2 stimulated with IL-2 for 48 hours produce large quantities of interferon-gamma (10,300 ± 1850 pg/ml) and modest amounts of tumor necrosis factor-alpha (200 ± 60 pg/ml). IL-4 and IL-10 secretion was below background levels. We then evaluated the cytolytic potential of KIL C.2 using a FACS-based flourolysis assay using GFP-labeled target cells. We observed that KIL C.2 potently lyse allogeneic A20 (H2d) lymphoma tumor targets without prior activation. In addition, KIL C.2 is moderately lytic against two syngeneic (H2b) targets: a myeloma cell line, 5T33MM, and BLL C.2, a tumorigenic leukemia/lymphoma cell line that was established from a WBM culture and has the CD19+CD25+CD43+sIg− pre-B-cell phenotype (S.T. and T.M.C., unpublished data). To study the mechanism of cytolysis we performed killing assays in the presence of anti-NKG2D (MI-6) or anti-NK1.1 (PK136) blocking antibodies. Blockade of either NK1.1 or NKG2D reduced cytotoxicity against BLL C.2 targets by 50%. BLL C.2 has the C57BL/6 background, thus does not express H60 but expresses the other NKG2D ligands Rae1 at high levels and Mult1 at moderate levels as determined by quantitative RT-PCR. In contrast, blocking NK1.1 and NKG2D had no effect on cytolysis of A20 suggesting the presence of alternative activation pathways such as those mediated by MHC class I alloantigen receptors. To examine the in vivo tumoricidal potential of KIL C.2, we used a C57BL/6 → BALB/c T-cell depleted (TCD)-WBM transplantation and A20-luciferase tumor model and monitored tumor progression using the Xenogen bioluminescence imaging system. BALB/c mice lethally irradiated and rescued with 2.5 × 107 TCD-WBM along with A20-luciferase inoculation uniformly died from tumor. Mice given TCD-WBM and 1 × 106 donor splenocytes did not develop tumor but died from lethal graft-versus-host disease (GVHD). In contrast, mice given TCD-WBM and two injections of KIL C.2 at doses of 1 × 107 per mouse were protected from tumor progression without developing signs of GVHD. These results show that KIL C.2 is a highly functional NK cell line with the capacity for proliferation, cytokine secretion, and cytotoxicity against syngeneic as well as allogeneic tumors. As such, KIL C.2 provides a useful cell line model for studies of NK cell biology and suggests a potentially effective strategy for NK cell-based immunotherapy.