bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mobilization Studies in Immunodeficient Mice Support a Role of Complement in Modulating the Trafficking of Hematopoietic Stem Cells (HSC) - A Pivotal Role of C3 Cleavage Fragments in Retention and C5 Fragments in Mobilization/Egress of HSC.
Authors: Reca Ryan , Wysoczynski Marcin , Magda Kucia , Janowska-Wieczorek Anna , Ratajczak Janina , Ratajczak Mariusz Z. 
Published: October 2019

Abstract
We postulate that mobilization of hematopoietic stem cells (HSC) is part of a immune-response and to support this notion i) complement (C) cascade cleavage fragments modulate trafficking of HSC (Blood2003;101,3784; Blood2004;103,2071) and ii) patients suffering from severe combined immunodeficiency (SCID) are poor mobilizers (Blood1996;88, 1104). To shed more light on mechanisms involved in C-mediated trafficking of HSC we studied mobilization in several strains of immune-deficient mice mobilized by employing various mobilization protocols. We noticed that mobilization of HSC is always preceded by C activation - occuring by the classical (G-CSF, CY) or alternative (polysaccharides, chemokines) activation pathways. Accordingly, the classical pathway of C activation after G-CSF or CY administration is a result of proteolytic modification of the BM environment leading to the exposure of a neo-antigen in BM tissue, and binding of natural IgM antibodies to this neo-epitope triggers activation of the C cascade. In support of this, IgM-deficient mice (SCID, RAG2null and Jh) are poor mobilizers and G-CSF-induced mobilization in these mice was restored after supplementation with purified C-activating inmmunoglobulins. On the other hand the mobilization in these mice (SCID, RAG2null and Jh) was normal in response to polysaccharides or chemokines that activate complement by the alternative IgM-independent pathway. To further support an important role of C in HSC mobilization, we studied HSC mobilization in mice deficient for various C components (C3−/− and C5−/− mice). We noticed that C3−/− mice are easy mobilizers. In contrast mobilization was very poor in C5−/− mice. This suggests that C3 and C5 cleavage fragments differently control the mobilization of HSC. We will present evidence that C3 cleavage fragments directly interact with HSC and prevent their uncontrolled egress from BM during mobilization by increasing their SDF-1-dependent marrow retention. In contrast C5 cleavage fragments activate neutrophils and BM-endothelium and thus are crucial for the egress of HSC. Thus the mobilization of HSC is i) dependent on C activation by the classical or alternative pathway, ii) modified differently by C3 cleavage fragments which enhance retention and C5 cleavage fragments which promote egress of HSC. Thus modulation of C activation in BM may help to develop new more efficient strategies for both HSC mobilization and their homing/engraftment.
Differential Expression of Type I MAGE in New and Relapsed Multiple Myeloma: Evidence for Association with Proliferation and Progression of Disease.
Authors: Cho Hearn J. , Ely Scott , Austin Wayne R. , Niesvizky Ruben , Pearse Roger , Coleman Morton , Bhardwaj Nina , DiLiberto Maurizio , Chen-Kiang Selina , Old Lloyd J. , Jungbluth Achim A. 
Published: October 2019

Abstract
The type I Melanoma Antigen GEne (MAGE) proteins belong to the Cancer-Testis family of tumor-associated antigens and are found in a broad range of solid and hematologic malignancies. We previously showed that the type I MAGE proteins CT7 (MAGE-C1) and MAGE-A3 were commonly detected in primary myeloma by both RT-PCR and immunohistochemistry (IHC). Higher levels of MAGE protein expression had a positive correlation with abnormally elevated proliferation as measured by the Plasma Cell Proliferation Index (PCPI, percentage of Ki-67+ cells in the CD138+ myeloma cell compartment). These findings suggest that MAGE may play a role in abnormal cell cycle regulation in myeloma. We explored this hypothesis by examining type I MAGE gene expression and proliferation by IHC in 46 newly-diagnosed, untreated and 35 relapsed myeloma patients, based on the clinical observation that relapsed patients exhibit lower response rates to therapy and shorter time to progression, indicative of more aggressive disease. PCPI was significantly higher in relapsed patients (19.0 ± 3.5%) compared to newly diagnosed (6.9 ± 1.3%, p<0.0002). Expression of CT7 and CT10 (MAGE-C2), a type I MAGE not previously associated with myeloma, was stable between newly-diagnosed and relapsed patients (76.0% of new samples vs. 77.1% of relapsed for CT7, 48.5% vs. 50.0% for CT10). In contrast, MAGE-A3 was detected in a significantly greater percentage of relapsed patients (77.1%) compared to newly diagnosed (35.6%, p=0.0003). The link between MAGE expression and unrestricted proliferation was further supported by in vitro studies with human myeloma cell lines. Proliferating myeloma cells were metabolically labeled with the nucleotide analog bromodeoxyuridine (BrdU) followed by intracellular staining and flow cytometry. This assay demonstrated that proliferating myeloma cells that incorporated BrdU into their genomic DNA expressed higher levels of type I MAGE protein compared to non-proliferating cells. Arresting cells at the G1-S interface by double thymidine blockade lead to the accumulation of cells expressing high levels of MAGE, which rapidly entered S phase and incorporated BrdU upon release from block. These results strongly suggest that expression of CT7 and CT10 are relatively early and stable events in the pathogenesis of myeloma, whereas activation of MAGE-A3 expression is associated with disease progression. Furthermore, MAGE expression is correlated with abnormal proliferation in vitro and in vivo, suggesting a potential functional role in the dysregulation of the cell cycle that is a hallmark of this disease. These results support the further exploration of the role of type I MAGE expression in myeloma and the development of therapeutic agents targeting them, especially tumor vaccines.
Targeting PKC in Multiple Myeloma: In Vitro and In Vivo Effects of the Novel, Orally Available Small-Molecule Inhibitor Enzastaurin (LY317615.HCl).
Authors: Raab Marc S. , Podar Klaus , Zhang Jing , McMillin Douglas , Breitkreutz Iris , Tai Yu-Tzu , Lin Boris , Ghobrial Irene , Munshi Nikhil , Hideshima Teru , Chauhan Dharminder , Anderson Kenneth C. 
Published: October 2019

Abstract
In multiple myeloma (MM), protein kinase C (PKC) overexpression has been reported including the prognostic adverse patient group with t (4;14) translocation. Importantly, PKC signaling pathways have been implicated in MM cell proliferation, apoptosis, and migration. Here, we investigated the novel, orally available PKC inhibitor Enzastaurin for its anti-MM activity. Enzastaurin specifically inhibits membrane, cytosolic, and nuclear phosphorylation of homologous PKC isoform residues, as well as associated kinase activity, induced by the major PKC activator TPA (Tumor-promoting phorbol ester). Consequently, it also abrogates TPA-induced phosphorylation of signaling molecules downstream of PKC including MARCKS and PKCm. In MM, Enzastaurin inhibits PKC activation triggered by growth factors and cytokines secreted by bone marrow stromal cells (BMSCs); co-stimulation with the extracellular matrix protein fibronectin, VEGF or IL-6; as well as MM patient serum. Phosphorylation of downstream signaling molecules was also abrogated, including cytoplasmic and nuclear ERK, JNK, ribosomal protein S6 and GSK3b as well as nuclear cMyc. Enzastaurin inhibits both proliferation and survival of MM cell lines and MM cells isolated from multidrug- resistant patients; as well as overcomes MM cell growth triggered by tumor cell binding to BMSCs and endothelial cells at a low micromolar range equivalent to the concentrations achieved in patient plasma during clinical trials. Importantly, synergistic cytotoxicity is observed when Enzastaurin is combined with bortezomib. Besides proliferation and survival, Enzastaurin inhibits MM cell adhesion, as well as VEGF- and IGF-1-triggered MM cell migration. It also blocks VEGF- triggered signaling pathways in endothelial cells, thereby inhibiting tubule formation. Finally and most importantly, tumor growth, survival, and angiogenesis are abrogated by Enzastaurin in an in vivo xenograft model of human MM. Our results therefore demonstrate in vitro and in vivo efficacy of the orally available PKC inhibitor Enzastaurin in MM and strongly support its clinical evaluation, alone or in combination therapies, to improve patient outcome in MM.
NSC 338258 Represents a Novel Anti-Neoplastic Agent for the Treatment of Multiple Myeloma.
Authors: Tian Erming , Landowski Terry , Stephens Owen , Yaccoby Shmuel , Barlogie Bart , Shaughnessy John D. 
Published: October 2019

Abstract
We have shown that high-risk multiple myeloma (MM) can be linked to amplification and overexpression of the cell cycle regulator CKS1B. With this in mind we correlated high expression of this gene in the NCI60 cell line panel with IC50 of over 10,000 anticancer compounds. NSC 338258 (EPED3) was identified in this search. EPED3 is a highly stable, hydrophilic derivative of ellipticine. In vitro, this synthetic compound exhibits drastic cytotoxic activity on myeloma cells, which is unique among most ellipticine derivatives. This efficacy of EPED3 was examined in myeloma cells co-cultured with bone marrow stromal cells. Despite the protective influence of stromal cells, micromolar concentrations of EPED3 were highly effective at killing myeloma cells; however, this lethal activity was exclusive of stromal cells. In co-culture, EPED3-induced cell cycle arrest and massive apoptotic progression appears to be a consequence of its instant impact on cytoplasmic organelles, particularly mitochondria. Disruption of mitochondrial and endoplasmic distribution of cytochrome c initiated the intracellular proteolytic cascade through the intrinsic apoptotic pathway. Effects of EPED3 treatment were further evaluated in myeloma cell lines with selective tolerances to doxorubicin, dexamethasone, and Velcade; EPED3 overcame these acquired drug resistances. In addition, the potency of EPED3 was tested on mononuclear cells isolated from peripheral blood of healthy donors. Under mitogenic stimulation, EPED3 had no significant growth inhibition effects within a range of concentrations killing myeloma cells. Collectively, our current data suggest that EPED3 is an extraordinary agent that, in vitro, targets mitochondrial function to rapidly deplete chemical energy and to initiate apoptosis in myeloma cells at low concentration, while leaving healthy stromal and mononuclear cells unharmed.
ARRY-797, a Potent and Selective Inhibitor of p38 Map Kinase, Inhibits LPS-Induced IL-6 and In Vivo Growth of RPMI-8226 Human Multiple Myeloma Cells.
Authors: Wright Dale , Winski Shannon L. , Anderson Deborah , Lee Patrice , Munson Mark , Winkler James 
Published: October 2019

Abstract
Multiple myeloma (MM) is characterized by the expansion of malignant plasma cells within the bone marrow. Their growth, survival, and migration are mediated in part via cytokines. Interleukin 6 (IL-6) is necessary for sustaining the in vitro growth of many MM cell lines and enhancing the proliferation of explanted human myeloma cells. The mitogen-activated protein kinase family member, p38, is activated by cytokines and growth factors and plays a significant role in inflammatory diseases. However, its role in the pathogenesis of multiple myeloma is poorly understood. Specific p38 inhibitors inhibit paracrine MM cell growth which is associated with IL-6 and VEGF secretion from bone marrow stromal cells (BMSCs). Furthermore, p38 inhibition blocks TNF-alpha-induced IL-6 secretion in BMSCs, thereby further inhibiting MM cell growth and survival. Although these data suggest an important role for p38 in MM, the direct effects of p38 inhibiton on MM has not been extensively explored. Therefore, we investigated the effects of p38 inhibition on in vitro and in vivo IL-6 production and MM cell growth in vivo after lipopolysaccaride (LPS) stimulation. LPS has been shown to induce various cytokines, including TNF-alpha and IL-6, via the p38 pathway. ARRY-797, an orally bioavailable, small molecule inhibitor of p38 directly inhibited LPS-induced IL-6 production from RPMI-8226 (IC50 = 100 pM) in vitro. In SCID-beige mice, LPS (3 μg/kg) induced IL-6 (7897 ± 827 pg/mL) and TNF-alpha (1922 ± 282 pg/mL) after 2 hours and these cytokines were inhibited by oral administration of ARRY-797 (30 mg/kg) by 91% and 95%, respectively. In MM xenograft models, ARRY-797 (30 mg/kg, BID, PO) inhibited RPMI 8226 tumor growth by 72% as a single agent and by 56% when LPS was administered to stimulate growth in vivo. In addition, ARRY-797 inhibited LPS-induced phosphorylation of p38 in RPMI-8226 xenografts. Together, these data support a role for p38 in IL-6-mediated growth of multiple myelomas. To our knowledge, ARRY-797 is the first small molecule p38 inhibitor to demonstrate single agent activity in a MM xenograft model and it has been advanced into preclinical development.
Heparanase Influences Expression and Shedding of Syndecan-1, and Its Expression by the Bone Marrow Environment Is a Bad Prognostic Factor in Multiple Myeloma.
Authors: Mahtouk Karene , Hose Dirk , Reme Thierry , Hundemer Michael , Jourdan Michel , Jourdan Eric , Pantesco Veronique , Baudard Marion , De Vos John , Larroque Marion , Moehler Thomas , Rossi Jean-Francois , Goldschmidt Hartmut , Klein Bernard 
Published: October 2019

Abstract
The heparan sulfate (HS) proteoglycan (PG) syndecan-1 plays a major role in multiple myeloma (MM), in part by concentrating HS-binding growth factors on the surface of MM cells (MMC). The function of HSPG is regulated by extracellular enzymes that modulate the structure of HS-chains. One such enzyme is heparanase (HPSE), an endo-glucuronidase that cleaves HS chains at only a few sites, resulting in fragments of 10–20 sugar units long that are biologically active.
Using Affymetrix microarrays and real-time RT-PCR, we showed that the gene encoding HPSE was expressed by 11/19 myeloma cell lines (HMCLs). In HSPE-positive HMCLs, syndecan-1 mRNA expression and production of soluble syndecan-1, unlike expression of membrane syndecan-1, were significantly increased. Downregulation of HPSE by HPSE-siRNA resulted in a decrease of syndecan-1 mRNA expression and soluble syndecan-1 production, without affecting membrane syndecan-1. Contrary to HMCLs, HPSE was expressed in only 4/39 primary MMC samples at a low level, whereas it was highly expressed in 36/39 whole bone marrow (WBM) microenvironment samples. In the latter, HPSE was expressed at a median level in polynuclear cells and T cells, at high level in monocytes and osteoclasts, and it was not expressed in BM stromal cells. In order to investigate the connection of clinical parameters with HPSE gene expression, 30/39 myeloma patients treated with high dose chemotherapy and autologous stem cell transplantation where classified into two groups: 15 patients with the lowest or the highest HPSE expression in the WBM. The HPSEhigh group had an increased percentage of patients with elevated C-reactive protein and β2-microglobulin. According to the international staging system (ISS), the HPSEhigh group included a significant higher frequency of patients with stage III MM and a lower frequency of patients with stage I MM. HPSEhigh patients had a shorter event free survival (EFS) (p=.017) and overall survival (OAS) (p=.023) than HPSElow patients. Classifying these 30 patients into two groups according to HPSE expression in MMC did not yield to any EFS or OAS significant differences between the two groups.
Altogether, those data provide evidence that HPSE not only modulate syndecan-1 activity through the cleavage of HS chains but also regulates syndecan-1 mRNA expression and shedding. We report for the first time that the expression of a gene in the BM, i.e. HSPE, is an indicator of poor prognostic for MM patients. Therefore, HPSE inhibitors like PI-88, which is currently in clinical trial, will be of interest for the treatment of patients with MM.
Cancer Germline Antigen-Specific T Cells Are Present and Functional in Many Patients with Multiple Myeloma and Paraproteinaemia.
Authors: Goodyear Oliver , Piper Karen , Khan Naeem , Starczynski Jane , Mahendra Prem , Pratt Guy , Moss Paul 
Published: October 2019

Abstract
The expression of Cancer Germline Antigens (CGAgs) is normally restricted to the pre-meiotic spermatogonia cells of the testis. The testis is an immunologically privileged site and so immunological tolerance to CGAg is not established. However, CGAg expression is also detected in many types of malignant disease including plasma cells from patients with multiple myeloma. CGAg expression has been shown to prime a T cell immune response in many patients with solid tumours and this may offer a novel target for immunotherapy in patients with myeloma.
We have used immunodominant peptide epitopes from a range of CGAgs to screen for CGAg-specific T cells in the blood of patients with multiple myeloma at various stages of their disease. Initial studies demonstrated that T cells from 15 out of 37 patients responded to one or more CGAg peptides and the magnitude of the CGAg-specific CD8+ T cell response ranged between 0.0004% and 0.1% of the total CD8+ T cell pool. Serial analysis showed that these immune responses were detectable in individual patients at multiple time-points during the course of their disease. A further 13 peptides have now been obtained including several CD4 peptide. We have subsequently cloned CD4 T cells specific to a MAGE 3 peptide and have shown them to be functional. In some patients we determined the membrane phenotype of the CGAg-reactive cells as CD45RA+ and CCR7−, an effector memory differentiation state. CGAg-specific responses have also been detected in patients with clinically benign forms of paraproteinaemia indicating that T cell immunity may play a role in the control of disease progression. Plasma cells are localised to bone marrow and we are now focussing on the study of immunity to CGAg at this site. Initial findings indicate a higher proportion of CGAg-specific T cells within bone marrow and the phenotypic profile of these cells is being determined. Functional T cells specific for CGAg are therefore present in a large proportion of patients with multiple myeloma and offer the possibility of a novel approach for immunotherapy in this disease.
Analysis of Varicella Zoster Virus Reactivation among Bortezomib-Treated Patients in the APEX Study.
Authors: Chanan-Khan Asher Alban , Sonneveld Pieter , Schuster Michael W. , Irwin David , Stadtmauer Edward A. , Facon Thierry , Harousseau Jean-Luc , Ben-Yehuda Dina , Lonial Sagar , Goldschmidt Hartmut , Reece Donna E. , Neuwirth Rachel B. , Anderson Kenneth C. , Richardson Paul G. 
Published: October 2019

Abstract
BACKGROUND: Bortezomib (btz, VELCADE®), a selective and reversible proteasome inhibitor, is approved for the second-line treatment of multiple myeloma (MM). There is a tendency for reactivation of varicella zoster virus (VZV;herpes zoster) among patients (pts) receiving immunosuppressive agents for cancer. Rates of VZV reactivation during btz therapy in phase 2 studies were reported to be 11% in SUMMIT and 13% in CREST. To investigate whether btz increases the incidence of VZV reactivation, data were reviewed from the phase 3 APEX trial of btz vs high-dose dexamethasone (dex) in pts with relapsed MM following 1–3 prior therapies.
METHODS: The APEX trial was described previously [Richardson et al. N Engl J Med2005;352:2487–98]. Use of prophylactic anti-virals and anti-bacterials was not mandated by this protocol.
RESULTS: Baseline characteristics, including age, prior ASCT, β2-microglobulin, and disease stage, were comparable between the btz and dex arms. The incidence of VZV reactivation was significantly higher with btz vs dex (13% [42/331] vs 5% [15/332], respectively; p=0.0002). Most VZV reactivation events in both arms were grade 1/2. Notably, the incidence of grade 3/4 events was similar between the two arms (1.8% [6/331] vs 1.5% [5/332]), as was the rate for serious adverse events (1.5% [5/331] vs 0.9%, [3/332], respectively). The incidence of other opportunistic infections, including VZV, was comparable between the btz and dex arms; 23% vs 20%, respectively. There were no deaths due to VZV reactivation in either arm. Median time to onset of VZV reactivation tended to be shorter in the btz arm vs dex, though this was not statistically significant (31 vs 51 days; p=0.221). At first assessment post VZV reactivation event, median ALC was significantly lower in the btz vs dex arm (1050 vs 1750 cells/μL; p=0.028). However, immediately prior to the event, median ALC was similar between arms (1030 vs 1100 cells/μL; p=0.529).
CONCLUSIONS: Btz was associated with a higher incidence of VZV reactivation events than dex. The reasons for this are unclear and require further study. Preliminary evidence from recent studies has shown that this adverse event of btz could be overcome with the use of prophylactic acyclovir [Mateos et al. Blood 2006; Epub ahead of print].
Lenolidamide (Revlimid), in Combination with Cyclophosphamide and Dexamethasone (CRD) Is an Effective Regimen for Heavily Pre-Treated Myeloma Patients.
Authors: Morgan Gareth J. , Schey Stephen , Wu Ping , Srikanth M. , Phekoo Karen J. , Jenner Matthew W. , Dines Sharon , Saso Radovan , Davies Faith E. 
Published: October 2019

Abstract
Lenolidamide (Revlimid®) is an oral immunomodulatory drug that has been shown to be effective for the treatment of relapsed refractory myeloma, and in-vitro laboratory studies suggest that its action may be synergistic with a number of conventional chemotherapeutic agents. The aim of this study is to assess the efficacy and toxicity profile when lenolidamide is used in combination with cyclophosphamide and dexamethasone for patients with relapsed refractory disease. Multiply relapsed patients were given Revlimid 25mg po on days 1–21, dexamethasone 40mg po days 1–4 and days 12–15, and cyclophosphamide 500 mg po days 1, 8, 15 and 21 of a 28 day cycle for a maximum of 6 cycles of treatment. Prophylaxis with acyclovir, septrin and proton pump inhibitors was routinely used, 1 high risk patient received prophylactic anticoagulation. Toxicity profiles and response were assessed every 4 weeks. To date 18 patients have been included in the study with median age of 60.5 years (range 34–76). All were heavily pre-treated with a median of 4 previous lines of therapies (range 2–8). 12 patients had received high dose melphalan, 18 patients thalidomide and 16 patients bortezomib. One case had undergone an allogeneic BMT. The median time from diagnosis to treatment initiation was 55.5 months (range 11–122). To date 81 complete courses of therapy have been given to 18 patients with median number of 5 courses (range 1–6). 8 patients experienced neutropenia with the neutophil count falling below 0.5×109/L in a total number of 14 cycles, which resulted in a dose reduction or stopping of cyclophosphamide in 8 patients. 12 patients received GCSF to maintain their neutrophil count. 5 patients required dose reduction/omit of Revlimid. 4 patients required intravenous antibiotics for neutropenic fever. The side effect profile was manageable, importantly no patient experienced sedation, constipation or worsening of peripheral neuropathy. 2 patients with a heavy myeloma load suffered a DVT, but continued on therapy achieving a PR once anti-coagulation had been commenced. 14 of the 17 patients assessable for response achieved a response with 1 CR, 3 VGPR, 8 PR and 2 MR according to EBMT criteria. The median time to response was prompt at 32 days (range 21–68). To date only 2 patients have discontinued therapy because of a failure to respond to therapy and 1 stopped due to liver toxicity. 13 patients have completed treatment and 5 still remain on treatment. The combination of CRD is effective in heavily pretreated myeloma patients and has a manageable toxicity profile. In view of the neutropenia further studies using this combination are currently being investigated using cyclophosphamide on days 1 and 8 only.