bluuebunny/crossref_metadata_2025_split huggingface dataset:
Osteosarcoma Derived from Cultured Mesenchymal Stem Cells.
Authors: Tolar Jakub , Nauta Alma J. , Osborn Mark J. , Panoskaltsis-Mortari Angela , McElmurry Ron T. , Bell Scott , Xia Lily , Zhou Ning , Riddle Megan , Schroeder Tania M. , Westendorf Jennifer J. , McIvor R. Scott , Hogendoom Pancras C.W. , Szuhai Karoly , Oseth LeAnn , Hirsch Betsy , Yant Stephen R. , Kay Mark A. , Peister Alexandra , Prockop Darwin J. , Fibbe Willem E. , Blazar Bruce R. 
Published: October 2019

Abstract
The beneficial effects of Mesenchymal Stem Cells (MSC) are being tested clinically in attempts to improve hematopoietic engraftment, to treat osteogenesis imperfecta, graft-versus-host disease and autoimmune diseases, and to deliver therapy for malignancies. In early reports, phase I clinical studies have not been associated with toxicities. To study the biodistribution of MSC, we labeled adult murine C57BL/6 MSC with firefly luciferase and DsRed2 fluorescent protein using non-viral Sleeping Beauty transposons, and co-infused labeled MSC with bone marrow into irradiated allogeneic recipients. Using in vivo whole body imaging, luciferase signals were shown to be increased between weeks 3 and 12. Unexpectedly, some mice with the highest luciferase signals died and all surviving mice developed foci of sarcoma in lungs. Two mice also developed sarcomas in their extremities. Infusion of MSC-derived sarcoma cells resulted in malignant lesions in secondary recipients. Common cytogenetic abnormalities were identified in tumor cells isolated from different animals. Mapping of the Sleeping Beauty transposition insertion sites did not identify an obvious transposon-related genetic abnormality. Importantly, the original MSC cultures not labeled with transposons, as well as cultured MSC independently isolated from the bone marrow of both BALB/c and C57BL/6 mice showed cytogenetic aberrations after several passages in vitro. Even though not all MSC cultures formed tumors upon in vivo injection, these data indicate that MSC transformation was neither strain-specific nor a rare event following ex-vivo expansion. Karyotype analyses using fluorescence in situ hybridization with spectral karyotyping (SKY) as well as combined binary ratio labeling of nucleic acid probes (COBRA) showed clonal evolution of transformed MSC suggesting that the critical transformation event(s) occurred before MSC infusion. Collectively, we describe cytogenetic instability of murine MSC isolated in two independent laboratories, their cellular transformation, and potential for sarcoma formation. While the growth characteristics of human and murine MSC are not identical and murine cells are more prone to undergo immortalization and transformation in culture than human cells, our study highlights the importance of quality control measures needed for ongoing and future clinical trials using human MSC.
Optimizing Mobilization, Augmentation, and Gene Transfer of Multipotent Mesenchymal Stromal Cells (MSCs).
Authors: Larsen Stephen R. , Chng Keefe , Zhou Shangzhen , Wright John , Armstrong Margaret , Thompson Sally , Hennessy Annemarie , Martiniello-Wilks Rosetta , Gibson John , Joshua Douglas E. , Rasko John E.J. 
Published: October 2019

Abstract
Aims: MSCs are cells being investigated for use in various therapies including facilitation of HSC transplantation and as gene therapy delivery vehicles. We have explored the potential to increase the number of bone marrow (BM) MSCs in vivo, induce mobilization using various cytokine regimens and improve gene transfer into these cells with adeno-associated virus (AAV) in a baboon model.
Method: Baboons received cytokines as follows: 1. G-CSF 100mcg/kg/day for 5 days; 2. pegylated G-CSF (pegG-CSF), single dose 300mcg/kg day −5; 3. G-CSF 100mcg/kg/day + stem cell factor (SCF) 50mcg/kg/day for 5 days; and 4. pegylated megakaryocyte growth and development factor (pegMGDF) 1mcg/kg second daily for 10 days + G-CSF 100mcg/kg/day for 5 days starting day −5. Animals underwent BM aspiration at baseline and on the final day of cytokines along with leukapheresis to isolate PBMNCs for detection of peripheral blood (PB) CFU-F. The immunophenotype and differentiation potential of CFU-F derived from animals before and after cytokines was compared. The ability of AAV vectors pseudotyped with capsids derived from AAV of serotypes 1, 2, 3, 4, 5, 6, and 8 to mediate transduction of baboon and human MSCs was assessed.
Results: Augmentation of bone marrow MSCs was observed with all cytokine regimens with the fold-increase compared to baseline as follows: 4.1, 2.1, 7.6 and 11.2 after G-CSF, pegG-CSF, G-CSF+SCF and G-CSF+pegMGDF respectively (see Figure 1). The immunophenotype of MSCs obtained after cytokines was identical to baseline cells as was their differentation potential. CFU-F were not detected in baseline PB however they were detected in 3/5 animals after G-CSF+SCF at a frequency of 0.8 to 1.5/mL, but no other cytokine regimen. A similar pattern of transduction efficiency using AAV was shared by human and baboon MSCs (see Figure 2) using control Ad293 cells. Specifically AAV vectors expressing capsids of serotypes 2, 3 and 5 were most efficient in transducing human and baboon MSCs. Those expressing capsids from serotypes 1, 4, 6, and 8 were much less efficient in transducing MSCs from either species. Baboon MSCs were able to be transduced by about 100-fold more than their human equivalent cells using AAV serotypes 2, 3, and 5.
Conclusion: This is the first report of mobilization of primate MSCs and, together with the demonstration of in vivo augmentation and AAV gene transfer, offers increased therapeutic opportunities for their safe application in a burgeoning number of diseases.
Figure 1: In vivo Bone Marrow CFU-F Augmentation following cytokines Figure 1:. In vivo Bone Marrow CFU-F Augmentation following cytokines Figure 2: Transduction profile of AAV vectors expressing capsids of various serotypes Figure 2:. Transduction profile of AAV vectors expressing capsids of various serotypes
The Novel HDAC Inhibitor UCL67022 Is Highly Potent in Multiple Myeloma and Non-Hodgkin’s Lymphoma and Is Enhanced by Bortezomib.
Authors: Maharaj Lenushka , Popat Rakesh , Chahwan Angela , Lister Andrew T. , Cavenagh James D. , Middleton Brian , Rioja Alf , Marson Charles , Joel Simon P. 
Published: October 2019

Abstract
Introduction: Histone deacetylases (HDACs) regulate the acetylation state of nucleosomal histones and represent a novel target in haematological malignancies. Inhibition of HDACs results in the induction of apoptosis in multiple myeloma (MM) and lymphoma (NHL) cells, associated with the down-regulation of signaling pathways involving IL-6 and IGF-1 and the inactivation of BCL-6. We have therefore investigated the activity of a novel hydroxamic acid HDAC inhibitor (UCL67022) in comparsion with SAHA, alone and in combination with the proteasome inhibitor bortezomib.
Methods: HDAC activity in partially purified rat liver homogenate and intact CEM cells was determined using a fluorescent HDAC substrate. The human MM cell lines RPMI 8226/S and U266, and the NHL cell lines SUD-4, CRL, DOHH2, DHL-4, 5, 6, 7, GRANTA-519 and JEKO-1 were used to investigate effects on viable cell number using an ATP-dependent bioluminescence method. Activity against primary malignant cells from patients with MM, DLBCL, FL or CLL was also studied. Western blot analysis was used to investigate changes in acetylated histone-H3 and a-tubulin.
Results: UCL67022 showed more potent HDAC inhibitory activity in liver extracts and whole CEM cells than SAHA (IC50 0.05 vs 0.39uM in liver and 0.11 vs 0.33uM in CEM cells). UCL67022 was more potent than SAHA in reducing viable cell number in U266 (EC50 0.14 vs 1.8 uM) and RPMI 8226/S (0.05 vs 0.78 uM) cells. Similarly NHL cell lines were 10–20 fold more sensitive to UCL67022, with median EC50 values of 0.05uM (range 0.03–0.10uM) vs 0.81uM, (range 0.68–1.28uM). In 2 primary MM samples UCL67022 showed increased activity with EC50 values of 0.7uM and 0.11uM vs 12.2uM and 1.1uM for SAHA. 3 FL patient samples and 1 CLL sample were 3.7-fold more sensitive to UCL67022 than SAHA (median EC50 4.7uM vs 17.5uM respectively). Western blot analysis showed a 10-fold difference in histone H3 acetylation between the two compounds, with acetylation returning to pre-treatment levels by 24hr with 3uM SAHA but remaining elevated out to 48hr with 0.3uM UCL67022. Increased acetylation of a-tubulin confirmed the inhibition of HDAC6. Combination with bortezomib at 1, 2 and 4nM in MM cells showed increased antiproliferative activity with both SAHA and UCL67022, with synergistic responses observed in the U226 cell line and in 1 primary sample and additive effects in the others.
Conclusions: These data demonstrate the activity of the highly potent, novel HDAC inhibitor UCL67022 in MM and NHL cell lines and primary patient cells. Activity was increased in co-exposures with bortezomib possibly due to inhibition of HDAC6 and subsequent aggresome formation, suggesting a therapeutic advantage with the combination.
FLT-PET in Haematological Disorders: A Novel Technique To Analyze the Proliferative Activity of the Bone Marrow Compartment.
Authors: Vellenga Edo , Agool Ali , Schot Bart , Jager Piet 
Published: October 2019

Abstract
Few diagnostic procedures are available to determine the degree of bone marrow cellularity and the numbers of cycling cells in patients with hematopoietic disorders. Therefore non-invasive imaging of the bone marrow compartment may be helpful. Recently the PET tracer [18F]-3′-fluoro-3′-deoxy-L-thymidine (FLT) has been developed. FLT uptake is related to the DNA synthesis rate and increases with higher proliferation rates and may therefore visualise the high cycling activity of haematopoietic cells in the bone marrow compartment. In the present study the feasibility of visualisation and quantification of the bone marrow compartment activity with FLT-PET was used to distinguish different haematological disorders.
Clinical and laboratory data of 18 patients with low-risk myelodysplasia (MDS), chronic myeloproliferative disorders, myelofibrosis, aplastic anemia or multiple myeloma were correlated with the results of FLT-PET imaging using visual analysis and SUV. Findings were compared with those in normal controls (n=14).
With SUV analysis of the femura, thoracic and lumbar vertebral body in conjunction with visual analysis a distinction could be made between MDS (n=9), chronic myeloproliferative disorders (n=3) and myelofibrosis (n=3) compared to normal controls (p<0.001). A significant increase in FLT uptake in the defined areas was observed in all the studied patients with MDS and myeloproliferative disorders. In addition a significant expansion of the bone marrow compartment was shown (p<0.001). No correlation was found between Ki-67 staining of bone marrow biopsies and FLT uptake. In contrast, patients with myelofibrosis demonstrated a decline in bone marrow FLT uptake compared to normal controls but a significant increase in liver and spleen uptake. Osteolytic lesions of multiple myeloma patients and patients with aplastic anemia showed a strong reduction in bone marrow FLT uptake.
The results indicate that FLT-PET can be used to visualize the proliferative activity of the bone marrow compartment and may be helpful to distinguish separate haematological disorders.
Hepcidin Expression Is Regulated by a Complex of Hemochromatosis-Associated Proteins.
Authors: Schmidt Paul J. , Huang Franklin W. , Wrighting Diedra M. , Toran Paul T. , Andrews Nancy C. 
Published: October 2019

Abstract
Hemochromatosis is a common genetic disease resulting from increased dietary iron absorption and tissue iron deposition. Mutations in five unrelated genes are known to cause hemochromatosis in humans and mice. These encode the classic hemochromatosis protein (HFE), transferrin receptor 2 (TFR2), the iron exporter ferroportin (FPN), hemojuvelin (HJV), and the circulating anti-microbial peptide hepcidin (HAMP). Hepcidin binds to FPN, causing its internalization and degradation, thus decreasing cellular iron release. A basic understanding of the pathophysiology of FPN and hepcidin mutations has recently been elucidated; however, it was still unclear how mutations in HFE, TFR2, and HJV cause hemochromatosis. All are associated with decreased hepcidin production and inappropriately high levels of ferroportin activity. HFE, TFR2 and HJV are normally expressed in the hepatic cells that produce hepcidin. With collaborators, we showed that HJV acts as a bone morphogenetic protein (BMP) co-receptor. HJV binds to the BMP ligands and forms a complex with Type I BMP receptors, resulting in signaling through a SMAD pathway and induction of hepcidin expression. Disease causing mutations in HJV abrogate BMP co-receptor activity, and hepatocytes from Hjv−/ − mice have a blunted response to BMP2. HFE was known to form a complex with the classical transferrin receptor, TFR1. Several models have been proposed implicating this complex in the regulation of normal iron homeostasis, but they have not taken the role of hepcidin into account. To examine the HFE/TFR1 interaction in vivo, we developed mice expressing a mutant form of TFR1 that should constitutively interact with HFE. We found that these transgenic animals have a phenotype similar to Hfe−/ − mice, suggesting that TFR1 serves to sequester HFE to silence its activity. We next asked whether HFE might also participate in BMP signaling. We found that forced expression of HFE in a hepatoma cell line induces transcription of a reporter gene linked to the hepcidin promoter. It also induces transcription from a heterologous promoter containing BMP-responsive elements, suggesting that HFE works through the BMP pathway. In contrast, forced expression of TFR2 did not amplify expression of either reporter, but it prevented cellular release of a soluble cleavage product of HJV. Furthermore, we showed that both HFE and TFR2 are associated with HJV in a stable protein complex that can be isolated by co-immunoprecipitation or Ni-affinity chromatography. TFR2 appears to aid in the recruitment of HFE to this complex. We conclude that HFE and TFR2 thus serve to amplify BMP signaling through an HJV/BMP receptor pathway. Our findings provide a compelling explanation for the similar clinical hemochromatosis phenotypes resulting from mutations in these genes.
Phase II Results of Bendamustine in Combination with Rituximab in Relapsed Indolent and Mantle-Cell Non-Hodgkin’s Lymphoma.
Authors: Van der Jagt Richard H. , Cohen Philip , Cheson Bruce D. , Tulpule Anil , Herst Jordan A. , Lemieux Bernard , Schwartzberg Lee S. , Robinson Katherine Sue , Oliver Jennifer W. , Williams Michael E. 
Published: October 2019

Abstract
Objective: The objective of this study was to evaluate the efficacy and safety of bendamustine HCl (TREANDA®) in combination with rituximab in patients with relapsed non-Hodgkin’s lymphoma (NHL).
Background: Bendamustine is a novel hybrid, alkylating agent with single-agent activity in multiple hematologic and solid tumors. It induces cell death via both apoptosis and the apoptosis-independent pathway of mitotic catastrophe. The combination of bendamustine and rituximab has been shown to exhibit a synergistic antitumor effect on NHL cells.
Methods: This Phase II, multicenter study enrolled adult patients with relapsed, indolent B-cell or mantle-cell NHL who were not refractory to rituximab (defined as progression ≤6 months of last rituximab dose). Patients received rituximab 375 mg/m2 intravenously (IV) on day 1 and bendamustine 90 mg/m2 IV on days 2 and 3 of a 28-day cycle for 4 to 6 cycles. An additional dose of rituximab 375 mg/m2 IV was given 1 week before the first cycle of bendamustine and 4 weeks after the last cycle.
Results: The intent-to-treat (ITT) population included 66 patients (59% men) with a median age of 60 years (range, 40–84). Indolent histologic phenotype was seen in 54 patients with the following histologic subtypes: follicular center cell (61%), small lymphocytic (15%), lymphoplasmacytic (3%), and marginal zone (3%); 18% had mantle-cell lymphoma (MCL). A total of 85% of patients had stage III/IV disease. These patients relapsed from a median of 1 prior chemotherapy (range: 0–5), with 56% having had prior treatment with rituximab. Patients with no prior chemotherapy relapsed following biologic therapy. In the ITT population, the overall objective response rate (ORR) was 94% (complete response [CR]/complete response unconfirmed [CRu], 41%; partial response [PR], 53%); 6% had stable disease. The ORR for the 12 MCL patients was 92% (CR/CRu, 42%; PR 50%). For all patients, the median duration of response and progression-free survival has not been reached after a median follow-up of 8.3 months (range, 0.14–31 months). Grade 3/4 neutropenia was seen in 41% of patients (7%, febrile neutropenia). Common nonhematologic toxicities (grade 1/2, grade 3, grade 4) were nausea (68%, 0%, 0%) and fatigue (53%, 5%, 0%); one patient had grade 3 sepsis. No alopecia was observed.
Conclusions: Bendamustine administered in combination with rituximab produced a high objective response rate and was generally well tolerated in patients with relapsed indolent and mantle-cell NHL who were not refractory to rituximab. These results suggest that the combination of bendamustine and rituximab may be comparable in activity to R-CHOP, and further studies of this combination are warranted.
Combination Immunotherapy with a CpG Oligonucleotide and Rituximab (R) Augments the Immunological Response and Favorably Alters the Malignant Microenvironment of Follicular Lymphoma (FL).
Authors: Friedberg Jonathan W. , Kelly Jennifer , Kutok Jeffrey L. , Salloum Rabih , Fisher David C. , Ronan Elizabeth , Dalton Virginia , Rich Lynn , Marquis Diana , Rothberg Paul G. , Liesveld Jane , Fisher Richard I. , Coffman Robert , Mosmann Tim , Freedman Arnold S. 
Published: October 2019

Abstract
The malignant microenvironment of FL, including T cells, macrophages and follicular dendritic cells, contributes to disease pathogenesis and represents a rational therapeutic target. Gene expression profiling and immunohistochemistry studies have confirmed that degree and type of immune response in the malignant FL lymph node impacts clinical outcome. CpG oligonucleotides have pleotropic effects on both the innate and adaptive immune systems, including increased antigen expression, enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) and shifting the immune response in a TH-1 direction. These agents therefore have significant potential to manipulate the immune response to FL, and alter the malignant microenvironment in a favorable way. We combined a CpG oligonucleotide (1018 ISS, 0.2 mg/kg sc weekly x 4 beginning day 8) with standard R in patients (pts) with relapsed/refractory, histologically confirmed FL, and comprehensively evaluated immunological changes following the combination. 22 pts (10 female; median age 55) with FL sensitive or naive to R were enrolled. FcγRIIIA polymorphism status was FF n=9, VF n=12, VV n=1. Treatment was well-tolerated with no significant adverse events attributable to therapy. Baseline and post therapy evaluations were performed on blood and the CpG injection sites in all pts. Compared to baseline, 7 pts demonstrated significant increases in ADCC after therapy using a conventional chromium-release assay. Circulating CD3-positive cells increased in 11 pts. Increased IFN-γrelease by Elispot occurred in 5 pts. 10 pts had significant increases in neutrophils, T cells, and macrophages in skin biopsies of CpG injection sites performed 24 hours post therapy compared with baseline. Pre- and post-biopsies of tumor tissue (including lymph nodes and bone marrow) were performed in 5 pts, and in all cases a significant infiltration of T cells (CD8 > CD4) and macrophages were observed following treatment. Clinical responses per Cheson criteria were observed in 45% of pts; median PFS was 18 months in responding pts, and 9 months for entire group; 12 pts have not yet required further therapy for FL. In conclusion, this group of pts with relapsed FL had favorable clinical outcome despite adverse prognostic factors. The addition of CpG to R had measurable and reproducible local and systemic immune effects in a substantial subset of pts. Unlike other cytokine/antibody combinations, this study is the first to histologically confirm perturbation of the local immune malignant microenvironment following systemic biological therapy of FL.
Prognostic Significance of Quantitative t(14;18) PCR Monitoring in Advanced Stage Follicular Lymphoma Patients.
Authors: Hirt Carsten , Schueler Frank , Kiefer Thomas , Haas Antje , Niederwieser Dietger , Neser Sabine , Assmann Michael , Dachselt Klaus , Leithaeuser Malte , Rabkin Charles S. , Herold Michael , Dolken Gottfried 
Published: October 2019

Abstract
In a prospective multicenter phase III trial (M 39023) advanced stage follicular lymphoma (FL) patients were randomized to receive eight cycles of MCP (mitoxantrone, chlorambucil, prednisolone) chemotherapy alone or in combination with rituximab (R-MCP). This study was carried out to determine the effects of this therapy on circulating lymphoma cells (CLC) and to assess the value of CLC quantitation as a molecular marker of disease activity and as a prognostic parameter. CLC numbers were determined by real-time quantitative PCR for the t(14;18)-MBR translocation or by allele-specific PCR for rearranged immunoglobulin heavy chain genes. Quantitative PCR of a reference gene (wild type K-ras) allowed the exact quantitation of cells tested per sample and to exclude samples with insufficient DNA content (< 500,000 cells). We analyzed serial blood samples from 43 patients obtained before, during and after completion of therapy. Baseline clinical characteristics and response to therapy of the 43 patients of this study were not significantly different to all 201 FL patients of the clinical trial except for a higher complete response (CR) rate in the R-MCP group (18/25 (72%) in this study versus 52/105 (50%) in the clinical trial, p=.04). Similar to the results of the clinical study, response rate and CR rate in the present study were significantly higher in the R-MCP arm than after therapy with MCP alone (25/25 (100%) versus 13/18 (72%), p= .009, and 18/25 (72%) versus 5/18 (28%), p= .006, respectively). Clearance of CLC at the end of therapy was achieved in 21/25 patients (84%) treated with R-MCP compared to 0/18 (0%) after MCP alone (p< .0001). R-MCP patients achieved a greater reduction of CLC after completion of therapy and a greater reduction of CLC per treatment cycle than patients treated with MCP alone, even if the comparison was restricted to patients with clinical response (3.88 log and 1.18 log reduction versus 2.21 log and 0.23 log reduction, p= .001 and p< .0001). A ≥ 2 log reduction of CLC after completion of therapy was associated with a favourable clinical response (p= 0.0007) and prolonged event-free survival (p= 0.02) regardless of treatment arm. Among patients with a ≥2 log CLC reduction there was no significant difference in EFS between patients with PCR negative samples and patients with PCR positive blood samples at completion of therapy (p= 0.091). In conclusion, R-MCP led to a rapid and sustained eradication of CLC in the majority of patients. The results of serial determinations of CLC numbers showed a good correlation with the quality and duration of the clinical response. Therefore, quantitative molecular disease monitoring could help to develop individualized treatment strategies for patients with advanced stage FL.