bluuebunny/crossref_metadata_2025_split huggingface dataset:
Myeloma with AL Amyloidosis: 10 Years Experience with Stem Cell Transplant.
Authors: De Padua Silva Leandro , Weber Donna , Wang Michael , Han Eric , Mendoza Floralyn , de Lima Marcos , Qazilbash Muzaffar , Champlin Richard E. , Giralt Sergio 
Published: October 2019

Abstract
Background: (AL) Amyloidosis is a rare and potentially fatal disease that involves deposition of light chain protein by a clonal plasma cell population. Treatment with high dose melphalan and autologous stem cell transplant (ASCT) can improve survival and reverse organ damage, but this treatment is associated with toxicity and mortality. We reviewed the outcome of patients with amyloidosis, who received hematopoietic stem cell transplant (HSCT) at our institution.
Patients and Methods: The retrospective study analyzed 32 patients with AL Amyloidosis who underwent transplant between 1997–2006. There were 17 men and 15 women, with a medium age of 53 years (range 35–73 years). All patients had diagnostic criteria for multiple myeloma (MM) and had biopsy confirmed amyloidosis of at least one organ site. Six patients had at least 1 organ involved with AL amyloidosis (3 bone marrows, 2 subcutaneous and 1 kidney), 20 patients had 2 organs involved, while 6 patients had 3 or more organs affected. All received a median of 3 cycles of chemotherapy (range 0–13) before transplant. Twenty-eight patients underwent autologous transplant, while 4 had an allogeneic transplants (2 syngeneic and 2 siblings). The preparative regimens received in 28 patients were high dose melphalan (26 autologous and 2 syngeneic), 2 patients received Busulfan and Melphalan, 2 allogeneic transplant Fludarabine and Melphalan.
Results: The median Charlson Comorbidities Index (CCI) was 3 (range 0–8). The median number of CD34+ cells infused was 4.85 x 106 cells/kg (range 1.43–7.82 cells/kg). The median time to neutrophil and platelet engraftment were 11 (range 2–21days) and 14 days (range2–41), respectively. 56% of patients developed moderate to severe gastrointestinal effects. Four patients underwent allogeneic transplant and 1 had acute and chronic graft vs host disease (GVHD). Twenty-one patients (65%) achieved a partial hematological remission (PR), and 5 patients (15%) achieved complete hematological remission (CR), with a total response hematological rate of 80%. The 100 day NRM was 3.2%. The median overall survival was (OS) 41 months (range 1–108), 7 patients died (4 relapsed and 3 infections) all 7 patients had CCI greater or equal to 4.
Conclusion: Selected patients with MM and amyloidosis can benefit from high dose therapy with stem cell support including allogeneic transplantation. The high mortality seen in patients with CCI >4 suggests that this index could be useful in selecting patients for high dose chemotherapy.
Mesenchymal Stromal Cells Gene-Enhanced To Express Human Factor IX and Their In Vivo Use for Correction of Hemophilia B Phenotype in R333Q Mice.
Authors: Coutu Daniel L. , Cuerquis Jessica , Griffith May , Blostein Mark D. , Galipeau Jacques 
Published: October 2019

Abstract
Hemophilia B is considered an appropriate disease target for gene therapy because it is a well characterized monogenic disease with a large therapeutic index. Despite promising preclinical and clinical trials in the last decade, safety and efficacy concerns associated with the in vivo administration of viral vectors still need to be addressed before gene therapy becomes part of the standard arsenal for clinicians. Our laboratory has developed a cell therapy approach using gene-enhanced autologous Mesenchymal Stromal Cells (MSCs) to deliver a therapeutic plasmatic protein which addresses these safety concerns. In this study, we tested whether MSCs engineered to express human Factor IX (hFIX) can be used to reverse the bleeding phenotype of R333Q hemophilia B mice developed by Stafford et al. We retrovirally engineered MSCs harvested from normal C57Bl/6 to express hFIX. A gene enhanced polyclonal population of MSCs was capable of producing carboxylated and fully active hFIX by in vitro clotting assays. By ELISA, the cells were shown to produce approximately 250ng of hFIX per million cells per 24h. Ten million of these cells were embedded in a collagen I gel matrix and implanted subcutaneously in R333Q hemophilia B mice (n=10). hFIX activity in mouse plasma (test and control groups) were followed weekly by aPTT assays. hFIX activity reached levels as high as 20% normal activity in some animals with an average +/− SEM of 11.2 +/− 2.1 (FIX activity in controls is <1%). The hFIX activity returned to baseline within 4 weeks. In conclusion, we demonstrate that gene-enhanced autologous MSCs can serve as an effective delivery of functional FIX for temporary correction of the hemophilia B phenotype. We hypothesize the presence of GFP co-expression by the implanted MSCs caused their immune rejection and we are currently testing this hypothesis.
Insertion of the 1.2kb Chicken β-Globin 5′HS4 Insulator Element in the LTR Inhibits HIV-1 Vector Mediated Transduction at the Early Infection Step.
Authors: Hanawa Hideki , Shimada Takashi 
Published: October 2019

Insertional oncogenesis is the major concern in gene therapy using integrating vectors such as gamma-retrovirus vectors and lentivirus vectors. One promising strategy to circumvent this unwanted event is the use of insulator elements which can block trans-activation of proto-oncogenes by enhancers of the integrated vectors. The 1.2kb fragment containing the chicken β-globin 5′ hypersensitivity site 4 (5′HS4) is the most well characterized insulator element. The insulator elements are usually inserted into the U3 region of the 3′ LTR of the vector construct, because the insulator element is duplicated and transferred to the 5′ LTR during viral replication. In the transduced cells, two insulator elements are located outside the retroviral transcriptional unit and this configuration is thought to be essential for efficient enhancer blocking. However, we have previously shown that insertion of the 1.2kb 5′HS4 element in the LTR of SIN-HIV-1 vector significantly reduced the transduction efficiency assayed by GFP expression in HeLa cells. In the present experiments, we examined the mechanism of reduced biological titer. We compared SIN-HIV-1 vectors with (INS) or without (PARENT) the 1.2 kb insulator element. The biological titer on HeLa cells (TU/ml), p24Ag concentration (ngp24/ml), and virus RNA genome content (vg copy/ngp24) of these vectors were shown in Table 1. As we previously reported, the biological titer of INS was significantly lower than that of PARENT, although p24 and RNA contents were not influenced by insertion of the 1.2 kb insulator element. In the next experiment, we performed reverse-transcription kinetic assay. Genomic DNA and low molecular weight DNA extracted by Hirt’s method were prepared from transduced HeLa cells and the amounts of reverse-transcribed vector sequence were determined by qPCR. The data showed that reverse-transcription of INS was remarkably reduced compared to PARENT. These results suggest that insertion of the 1.2 kb insulator does not inhibit production of virus particles and packaging of RNA genome. The responsible step for reduced titer of INS appears to be somewhere from the cellular entry step to the reverse-transcription step.
Characterization of HIV-1 vector with the 1.2 kb 5′HS4 insulator
x106 TU /ml ngp24 /ml x103 vg copy/ngp24 Values are mean ± sem from quadruplicate. a: p<0.005 PARENT 27 ± 4.3 361 ± 56 860 ± 16 INS 4.8 ± 0.4 a 348 ± 37 650 ± 81
Adenovirus-Mediated Gene Transfer to Mesenchymal Stem Cells: Comparison of the Cytomegalovirus- and Rous Sarcoma Virus-Promoter.
Authors: Deng Weiwen , Giambernardi Troy A. , Hay Rick V. , Pietryga Daniel W. , Abdel-Mageed Aly S. 
Published: October 2019

Abstract
We have compared the efficacy and toxicity of the cytomegalovirus (CMV) promoter and the Rous sarcoma virus (RSV) promoter for expressing genes for adenovirus-mediated gene transfer to mesenchymal stem cells (MSCs). Mouse MSCs (mMSCs) were isolated from the femurs and tibias of 6-week old, female BALB/c mice and ex vivo expanded in MEM-alpha containing 20% fetal bovine serum, 100 u/ml penicillin, 100 μg/ml streptomycin, 250 ng/ml amphotericin B, and 2 mM L-glutamine. To transduce mMSCs with adenovirus, the cells were plated at a density of 10,000 cells/cm2 in 6-well plates and cultured overnight. The cells were counted and then exposed to fresh culture medium containing adenovirus at 0 to 2,000 multiplicities of infection (MOI) for 48 h. Two adenoviral vectors with CMV promoter (Adv/CMV) and two adenoviral vectors with RSV promoter (Adv/RSV) were used in this study. Transduction efficiency and cell survival were then determined. The Adv/CMVs, i.e. AdvCMVlacZ and AdvCMVIL-10, were more potent in terms of transduction efficiency and transgene expression than their corresponding Adv/RSVs, i.e. AdvRSVlacZ and AdvRSVIL-10. However, both vectors exhibit a dose-dependent relationship between the adenoviral vector MOI and the percentage of transgene-expressing cells. Furthermore, no toxicity was observed in Adv/CMV or Adv/RSV transduced mMSCs.
Human Herpesvirus 8 K1 Blocks Fas in Cells and Mice by Binding and Blocking Fas.
Authors: Maeng Hoyoung , Wang Suizhao , Berkova Zuzuna , Wang Shu , Jung Jaehoon , Felipe Samaniego 
Published: October 2019

Abstract
Background: Kaposi’s sarcoma and primary effusion lymphoma are linked to human herpesvirus 8 (HHV-8 or Kaposi’s sarcoma-associated virus) infection. Key to the pathogenesis of these cancers is the HHV-8 transmembrane oncoprotein K1. This viral protein, which immortalizes cells, produces lymphoproliferation and lymphomas through unknown mechanisms when expressed in mice.
Results: We show here that K1 blocks Fas (CD95)-mediated apoptosis by binding the ectodomain of K1 to Fas in hematopoietic cancer cells (BJAB, U937 and THP-1). K1 bound to Fas in K1-transfected BJAB cells. To assess whether K1 and Fas formed complexes in cells, we chemically crosslinked proteins in transfected cells using 3,3′-dithiobissulfosuccinimidyl propionate in the presence of N-ethyl maleimide. Stabled K1-Fas complexes were present in intact cells despite blocking of free reactive cysteines. The selective ability of K1 to antagonize Fas-mediated apoptosis was strictly dependent on having its immunoreceptor tyrosine-based activation motif (ITAM). Using activating anti-Fas (CH-11) and nonactivating anti-Fas (B10) antibodies in a serial manner, we showed that K1 associated with Fas that was not activated, strongly suggesting that K1 prevents Fas from being activated. Mice transfected with K1 were protected from the apoptosis inducing effects of agonistic Jo2 anti-Fas antibody and tissues from these mice contained K1-Fas complexes. K1-transfected compared to vector-transfected mice had fewer terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling positive cells (13 ± 3% vs. 60 ± 7%, P=0.008), fewer activated caspase 3 positive cells (24 ± 5% vs. 88 ± 6%, P=0.006). Tissues had less hemorrhaging, and necrosis.
Conclusion: These data support a mechanism of K1 direct binding to and blockage by K1 to Fas in suppression of apoptosis in an ITAM-dependent manner, which may be a key step in lymphoma development.
A Cost-Utility Analysis of Nilotinib for CML Patients Who Have Become Resistant to Imatinib.
Authors: Pilgrim H. , Jewitt K. , Ward S. 
Published: October 2019

Abstract
Background: New generation BCR-ABL tyrosine kinase inhibitors are promising treatments for CML patients who are imatinib resistant or intolerant. The objective of this study was to consider the cost-utility of nilotinib in comparison to ‘high dose’ (800 mg) imatinib (the current standard comparator in the UK) for chronic-phase CML patients resistant to standard dose 400 mg imatinib using a lifetime horizon. The analysis is based on a UK NHS perspective.
Methods: A Markov model was developed using interim data from the phase II CAMN107A2101 registration study of nilotinib and data from the 39 patients who were dose-escalated, due to a sub-optimal response, to 800 mg imatinib within the IRIS phase III trial of imatinib versus interferon. Based on this evidence, the estimated 18-month survival has been extrapolated over a lifetime using parametric Weibull regression. Since the price of nilotinib in the UK was still to be determined at the time of preparation, the cost was assumed to be £106.94 per day, price parity with high dose imatinib. The costs and utilities associated with adverse events have been incorporated during the first six months following treatment. All costs and utilities were discounted at 3.5%.
Results: Nilotinib is estimated to cost an additional £49K and generate an additional 2.18 Quality-Adjusted Life Year’s (QALYs) in comparison to high dose imatinib over a patient’s lifetime. Therefore, the incremental cost-utility of nilotinib versus high dose imatinib is expected to be approximately £22K per QALY gained. This result is subject to uncertainty around data extrapolation. However availability of longer term data from the ongoing nilotinib trial will allow validation at a later date.
Conclusions: Our analysis suggests that nilotinib provides a clinically and cost-effective treatment (according to conventionally acceptable thresholds) for CML patients who have become imatinib resistant, in an indication where limited alternative treatment options exist.
Prognostic Significance of VEGF, VEGF Receptors, and Microvessel Density in Diffuse Large B Cell Lymphoma Treated with Anthracycline-Based Chemotherapy.
Authors: Gratzinger Dita , Zhao Shuchun , Tibshirani Robert J. , Hsi Eric D. , Hans Christine P. , Pohlman Brad , Bast Martin , Avigdor Abraham , Schiby Ginette , Nagler Arnon , Byrne Gerald E. , Lossos Izidore S. , Natkunam Yasodha 
Published: October 2019

Abstract
Vascular endothelial growth factor (VEGF)-mediated signaling has at least two potential roles in diffuse large B cell lymphoma: potentiation of angiogenesis, and potentiation of proliferation and/or survival induced by autocrine VEGF receptor-mediated signaling by lymphoma cells. We have recently shown that diffuse large B cell lymphomas expressing high levels of VEGF protein also express high levels of VEGF receptors 1 and 2. We have now assessed a larger multi-institutional cohort of patients with de novo diffuse large B cell lymphoma treated with anthracycline-based therapy to address whether either tumor vascularity as assessed by microvessel density counting, or expression of VEGF protein and its receptors as assessed by immunohistochemistry, contribute to patient outcomes. Increased tumor vascularity (Figure 1a, b, c -- increasing microvessel density) is associated with poor overall survival (p=0.047), and is independent of the international prognostic index (Figure 1d).
In contrast, high expression of VEGFR1 by lymphoma cells is associated with improved overall survival (p=0.044) and a trend toward improved progression-free survival (p=0.054). High expression of VEGF by lymphoma cells is also associated with a trend toward improved overall survival (p=0.089). The concordant trend toward improved survival with increased lymphoma cell expression of VEGF and its receptor VEGFR1, together with their coordinate overexpression in a subset of diffuse large B cell lymphomas (p=0.00025, chi value 21.5), suggests the presence of an autocrine VEGF-VEGFR1-mediated feedback loop in these lymphomas. Indeed the combination of high VEGF and VEGFR1 protein expression identifies a subgroup of patients with improved overall (p=0.003) and progression-free (p=0.026) survival; these findings are independent of the international prognostic index (Figure 2).
The presence of improved survival in a cohort of patients whose lymphomas potentially depend on autocrine signaling via VEGFR1 suggests that dependence on this pathway may render patients susceptible to the effects of anthracycline-based therapy; indeed, downregulation of VEGF expression at the mRNA level by anthracyclines and other chemotherapeutic agents has been demonstrated in a variety of non-hematolymphoid neoplasms. The relative importance of autocrine VEGF-mediated signaling versus vascularity should certainly be taken into account in clinical trials of anti-VEGF/VEGF receptor therapy in diffuse large B cell lymphoma.
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SB1518: A Potent and Orally Active JAK2 Inhibitor for the Treatment of Myeloproliferative Disorders.
Authors: Goh Kee C. , Ong Wai C. , Hu Changyong , Hentze Hannes , Liang Ai L. , Stunkel Walter , Tan Yong C. , Sangthongpitag Kanda , Goh Siok K. , Sieh Shirly , Bonday Zahid Q. , William Anthony D. , Lee Angeline , Blanchard Stephanie , Liu Te C. , Sattler Martin , Griffin James D. , Dymock Brian W. , Kantharaj Ethirajulu , Wood Jeanette M. 
Published: October 2019

Abstract
JAK2 is the most common mutated gene in bcr-abl-negative chronic myeloproliferative disorders (MPDs) making it an attractive target for drug discovery. Pre-clinical studies have shown that inhibition of JAK2 signaling pathways confer therapeutic benefits in animal models of MPD. We describe here SB1518, a potent, selective and orally active inhibitor of JAK2 with therapeutic potential for the treatment of MPDs. SB1518 is a potent ATP-competitive inhibitor of both JAK2 kinase (IC50 = 22 nM) and its JAK2V617F mutant (IC50 = 19 nM) without significant activity against 40 other representative kinases across the human kinome. SB1518 selectively inhibits the proliferation of cell lines driven by JAK2 or its mutants (e.g. IC50 = 81 nM for a murine BaF3 cell line stably transfected to express erythropoietin receptor and GFP-labeled JAK2V617F). It reduces basal phosphorylation of JAK2 and STAT5 in BaF3-JAK2V617F cells in a concentration-dependent manner. The excellent physicochemical, metabolic and pharmacokinetic properties of SB1518 render it amenable to oral dosing. SB1518 was investigated in a model of advanced MPD established by intravenous injection of BaF3-JAK2V617F cells in nude mice. In several studies using different initial tumor burden and/or drug regimen, we observed dose-dependent and statistically significant therapeutic effects including normalization of elevated white blood cell count and reduction of GFP-labeled BaF3 cells in the peripheral blood, resolution of hepatosplenomegaly, reduction of phospho-STAT5 in diseased organs, prolonged survival and alleviation of terminal-stage anemia and thrombocytopenia. These therapeutic effects were observed at doses well tolerated by the animals. In conclusion, our data demonstrate the therapeutic potential of SB1518 for the treatment of myeloproliferative disorders caused by aberrant JAK2 signaling.
Significant Role of Peptidyl-Prolyl cis/trans Isomerase, Pin1 in Acute Myeloid Leukemia with C/EBPα Mutations.
Authors: Pulikkan John Anto , Dengler Viola , Peer Zada Abdul A. , Gelutu Mulu , Tenen Daniel G. , Behre Gerhard 
Published: October 2019

Abstract
Transcription factor CCAAT enhancer binding protein α (C/EBPα) is crucial for the differentiation of granulocytes. Experimental data from animal models as well as patient samples suggest that loss of function or expression of C/EBPα provides a platform on which acute myeloid leukemia (AML) develops. C/EBPα is mutated in around 9% of acute myeloid leukemia. The mutations reported in C/EBPα are frame shift mutations at N-terminal domain and point mutations at basic region Leucine zipper. The mutant form of C/EBPα ie C/EBPα-p30 exhibits dominant negative function over the wild type protein. Peptidyl-prolyl cis/trans isomerase, Pin1 binds to and isomerizes the peptidyl-prolyl bond in specific phosphorylated Ser/Thr-Pro motifs. A growing number of studies show that Pin1 is overexpressed in many cancers and has significant role in tumorigenesis. In the present study we investigated the role of Pin1 in acute myeloid leukemia with C/EBPα mutation. Here we report C/EBPα-p30 could induce Pin1 transcription as assessed by quantitative Real-Time RT-PCR analysis. Affymetrix mRNA expression analysis show that Pin1 is upregulated in patients with acute myeloid leukemia. Silencing of Pin1 could overcome the dominant negative action of the C/EBPα-p30 over the C/EBPα-p42 transactivation capacity as analyzed by promoter assay. By silencing Pin1 with inhibitor against Pin1 (PiB), we could show myeloid differentiation in Kasumi-6 cells by FACS analysis with granulocytic specific markers. Western blot analysis shows that Pin1 inhibition by PiB could upregulate wild type C/EBPα protein level. Luciferase promoter assay for the Pin1 promoter shows that C/EBPα-p30 induces Pin1 promoter activity in association with E2F1. Mean while, wild type C/EBPα interferes with transactivation of the Pin1 promoter and downregulates Pin1 mRNA expression. We investigated the mechanism underlying the dominant negative action of C/EBPα-p30 over the wild type protein. We have previously shown that c-Jun expression is high in AML patients with C/EBPα mutation and c-Jun could block C/EBPα function by protein-protein interaction. Quantitative Real-Time RT-PCR analysis shows that overexpression of Pin1 induces c-Jun mRNA expression, while inhibition of Pin1 by the inhibitor PiB downregulates c-Jun mRNA expression. We show that c-Jun blocks the DNA binding and transactivation of wild type C/EBPα protein as assessed by gel shift assay and promoter assay respectively. In conclusion, inhibition of Pin1 leads to granulocytic differentiation of human myeloid cells. Our findings suggest inhibition of Pin1 as a novel strategy in treating AML patients with C/EBPα mutation.