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1
Wider, Tobias. "Bausteine der Vertriebssteuerung bei DHL Express." Sales Excellence 22, no. 7 (August 2013): 26–35. http://dx.doi.org/10.1365/s35141-013-0729-8.
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Coltman, Tim, John Gattorna, and Stuart Whiting. "Realigning Service Operations Strategy at DHL Express." Interfaces 40, no. 3 (June 2010): 175–83. http://dx.doi.org/10.1287/inte.1100.0491.
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Angeline, Chriesty, and Siti Nurbaiti. "Tanggung Jawab Perusahaan Pengangkut DHL Express Terhadap Pengiriman Barang dari Jakarta ke Malang ( Studi Putusan Pengadilan Nomor 733/Pdt.G/2017/PN.JKT.SEL.)." Jurnal Hukum Adigama 2, no. 1 (July 23, 2019): 406. http://dx.doi.org/10.24912/adigama.v2i1.5246.
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In a transportation agreement, practice the rights and obligations of the parties are not always fulfilled, because during the process of shipping, sometimes it does not always went well, even there is the package disappearance cases. For example, a case that occurred between the DHL Express transport company and the sender Massayu Chairani who was disadvantage due to the loss of her package that the company agreed to delivered. How the company DHL Express responsibility of the shipping to the sender in transportation from Jakarta to Malang and How the District Court Decision Number 733 / Pdt.G / 2017 / PN.JKT.SEL regarding the responsibility of the shipping company DHL Express to the sender in transportation from Jakarta to Malang continue to make a discussion. The research method used is descriptive normative legal research method, using secondary data and primary data as supporting data with the law approach. The results of research illustrate that DHL Express does not give full responsibility to the sender and the results of judges' decisions that do not grant full compensation claims are also considered not in accordance with Article 91 KUHD and Law Number 22 of 2009 concerning Road Traffic and Transportation in Article 188 and Article 193 paragraph (1). It is recommended that DHL Express give full responsibility to the sender of the goods for transporting goods from Jakarta to Malang and should have a court decision can decide the case more carefully to grant full compensation claims.
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Литвиненко, І., and Руссом Абрам Ерімас. "DIGITAL TRANSFORMATION OF CONSUMERS: STUDY ON THE BASE OF DHL EXPRESS IN ERITREA." Підприємництво та інновації, no. 8 (December 30, 2019): 62–68. http://dx.doi.org/10.37320/2415-3583/8.10.
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Digital innovation starts with a problem worth solving. We’ve found it’s amazingly helpful to have a single statement that represents the nature of the problem, which’s experiencing it, and why it’s so important to solve. There for the main problem is “To what extent consumers are fitted and accepted Digital transformation attributes”. This research could serve as a base for further investigation and improvement on the DHL express in Eritrea. Enhance our understanding of digital system and consumers driving. Moreover, it will contribute to the understanding of whether consumer are accepted and satisfied the digital transformation system in a developing country context. This research could also serve as a base for anyone who wants to study about how much consumers are accepted the digital transformation driving on them without any compliment on Postal Express service delivered in Eritrea. In addition, it will create a sense of awareness to the people who are engaged in offering digital system services on DHL. To undertake this research paper, primary and secondary data used. Primary data gathered by distributing online questionnaire to a sample of consumers. Secondary data extracted from several sources such as internet, magazine, books, periodicals, journals, company documents, etc. This research focuses on the digital transformation of consumers on DHL express despite the fact that consumer driving in digital transformation system is critical issue in any type of business. It focuses only on transformation from analog to digital system on consumers in experience on DHL express. It doesn’t concern about the technical issues of the DHL express over the entire world.
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Možuchová, Lucia, and Denisa Obermajerová. "APLIKÁCIA DIAGNOSTICKEJ METÓDY CRITICAL TO QUALITY V POŠTOVOM PODNIKU DHL EXPRESS." Pošta, Telekomunikácie a Elektronický obchod 11, no. 1 (2016): 27–35. http://dx.doi.org/10.26552/pte.c.2016.1.5.
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Osti, Cristoforo. "DHL Express (Italy) v Commission: Guidance on Parallel Immunity/Leniency Applications." Journal of European Competition Law & Practice 7, no. 7 (April 29, 2016): 460–61. http://dx.doi.org/10.1093/jeclap/lpw027.
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Suljagic, Mirza, Luca Laurenti, Muhammad Alam, Pablo G. Longo, Sami N. Malek, and Dimitar G. Efremov. "Chronic Lymphocytic Leukemia B-Cells Lack or Express Markedly Reduced Levels of the Phosphatase PHLPP, Which Regulates B-Cell Receptor Induced AKT Activation." Blood 112, no. 11 (November 16, 2008): 2080. http://dx.doi.org/10.1182/blood.v112.11.2080.2080.
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Abstract The PI3K/AKT pathway plays a central role in regulating cellular growth and survival. This pathway is activated by signals derived from various receptors and is tightly regulated through the action of several phosphatases, including SHIP and PTEN, which hydrolyze the PI3K product PIP3, and the recently identified PHLPP, which directly dephosphorylates AKT. Hyperactivation of the PI3K/AKT pathway has been implicated in the pathogenesis of many types of cancer, including chronic lymphocytic leukemia (CLL) and B-cell lymphoma. In addition, gene expression profiling and real-time RT/PCR analysis have recently shown differential expression of PHLPP mRNA in CLL subsets classified according to the presence of the 13q14 abnormality, with many CLL cases demonstrating absent PHLPP expression altogether. These findings prompted us to compare the levels of PHLPP expression in primary CLL B-cells (n=17) with normal tonsillar B-cells (n=4) and various lymphoma cell lines, including the diffuse large B-cell lymphomas (DLBCL) DHL-4, DHL-6, DHL-8, DHL-10, WSU, Toledo, Ly1, Ly3, Ly7 and Ly18, the Burkitt’s lymphoma BJAB and the prolymphocytic leukemia MEC1. Immunoblotting analysis revealed abundant and uniform expression of PHLPP in normal B-cells and in 7 out of 12 investigated lymphoma cell lines. Higher levels were observed in the BJAB, Ly1 and Ly18 cell lines, whereas PHLPP was undetectable in the DLBCL cell lines WSU and Toledo. Remarkably, PHLPP was either not expressed or was expressed at markedly reduced levels in all of the investigated CLL samples, with levels of expression ranging from 0 to 10% of the levels in normal B-cells. In contrast, the levels of expression of the phosphatase SHIP were relatively similar between CLL and normal B-cells. To determine what are the consequences of reduced PHLPP expression on signaling through AKT in malignant B-lymphocytes, we downregulated PHLPP in BJAB and DHL-4 cells by RNA interference. A significant reduction in the levels of PHLPP was achieved in both cell lines, which amounted to 20–40% of the levels in cells transfected with the control siRNA. Immunoblotting analysis of protein extracts from cells transfected with PHLPP and control siRNA did not show a difference in AKT phosphorylation on Ser473 and Thr308, indicating that a reduction in PHLPP expression is not sufficient to augment basal AKT activity. To determine the effects of PHLPP downregulation on agonist-induced AKT activation, we investigated phosphorylation on Ser473 and Thr308 in BJAB and DHL-4 cells stimulated through the B-cell receptor. In both cell lines downregulation of PHLPP resulted in more than a 50% increase in BCR-induced AKT phosphorylation. In contrast, phosphorylation of other signaling molecules that are also activated by BCR crosslinking, such as PLCγ2 and ERK, appeared unaffected by PHLPP downregulation. These data confirm the functional relevance of PHLPP in AKT regulation in B-lymphoid cells and implicate reduced or absent PHLPP expression in CLL B-cells as a potential determinant of BCR-induced AKT signaling in CLL.
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Wirtz, Jochen, Indranil Sen, and Sanjay Singh. "Customer asset management at DHL in Asia." Emerald Emerging Markets Case Studies 1, no. 1 (January 1, 2011): 1–6. http://dx.doi.org/10.1108/20450621111117413.
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Subject area Marketing; customer segmentation; operations and logistics. Study level/applicability Undergraduate business and management students, MBA/MA level application for international marketing modules incorporating customer segmentation and customer asset management. Case overview DHL, the international air express and logistics company, serves a wide range of customers, from global enterprises with sophisticated and high volume supply-chain solutions shipping anything from spare parts to documents, to the occasional customer who ships the odd one or two documents a year. To be able to effectively manage such a diverse customer base, DHL implemented a sophisticated customer segmentation cum loyalty management system. The focus of this system is to assess the profitability from its customers, reduce customer churn, and increase DHL's share of shipments. Expected learning outcomes Case teaching objectives: to demonstrate the concept of customer segmentation with loyalty management as a total system in a logistics company setting, and to evaluate appropriateness of the classification; to utilize the concept of service tier model within the company's current operations, and to evaluate the effectiveness of the model; to analyze the implementation of the customer segmentation cum loyalty management system and development of the necessary rules required to classify the various accounts into categories; to highlight the possible challenges arising from the implementation of customer segmentation cum loyalty management system, and to discuss possible methods of resolution. Supplementary materials Teaching note.
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Hideshima, Teru, Dharminder Chauhan, Kenji Ishitsuka, Noopur Raje, Shaji Kumar, Klaus Podar, Constantine Mitsiades, et al. "Molecular Characterization of PS-341 (bortezomib) Resistance: Implications for Overcoming Resistance Using Lysophosphatidic Acid Acyltransferase (LPAAT)-β Inhibitors." Blood 104, no. 11 (November 16, 2004): 2411. http://dx.doi.org/10.1182/blood.v104.11.2411.2411.
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Abstract PS-341 (bortezomib, Velcade™) is a promising novel agent for treatment of advanced multiple myeloma (MM); however, 65% of patients with relapsed refractory disease in a phase II study did not respond. Lysophosphatidic acid (LPA) is a phospholipid which mediates tumor cell migration and invasion. Recent studies have shown that inhibition of LPAAT-β inhibits both Ras/Raf/Erk and PI3K/Akt signaling cascades. We have previously shown that lysophosphatidic acid acyltransferase (LPAAT)-β inhibitor CT-32615 triggers caspase-dependent apoptosis, and can overcome resistance to conventional therapeutics (ie, dexamethasone, doxorubicin, melphalan) in MM cells. In this study, we determined whether CT-32615 could also overcome resistance to PS-341. We first characterized molecular mechanisms of resistance to PS-341 in DHL-4 lymphoma cells. DHL-4 cells express low levels of caspase-3 and caspase-8; furthermore, no cleavage in caspase-8, caspase-9, caspase-3, poly ADP-ribose polymerase (PARP), or DNA fragmentation factor (DFF) 45 is triggered by PS-341 treatment. We have previously shown that PS-341 treatment triggers phosphorylation of c-Jun NH2-terminal kinase (JNK), which subsequently induces caspase-dependent apoptosis; conversely, JNK inhibition blocks PS-341-induced apoptosis. Here we show that PS-341 does not induce phosphorylation of SEK-1, JNK, and c-Jun in DHL-4 cells, suggesting that it does not trigger a stress response. Importantly, CT-32615 inhibits growth of DHL-4 cells in a time- and dose-dependent fashion: a transient G2/M cell cycle arrest induced by CT-32615 is mediated via downregulation of cdc25c and cdc2. CT-32615 triggers swelling and cell membrane destruction in DHL-4 cells, without caspase/PARP cleavage or TUNEL-positivity, suggesting a necrotic response. Our studies therefore demonstrate that LPAAT-β inhibitor CT-32615 triggers necrosis even in PS-341-resistant DHL-4 cells, providing the framework for its evaluation to overcome clinical PS-341 resistance and improve patient outcome.
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Dougherty, G. J., P. M. Landorp, D. L. Cooper, and R. K. Humphries. "Molecular cloning of CD44R1 and CD44R2, two novel isoforms of the human CD44 lymphocyte "homing" receptor expressed by hemopoietic cells." Journal of Experimental Medicine 174, no. 1 (July 1, 1991): 1–5. http://dx.doi.org/10.1084/jem.174.1.1.
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In addition to the 85-95 kD CD44 species found on most hemopoietic cell types, the human myelomonocytic cell line KG1a expresses proteins of approximately 115 kD and 130 kD that react with monoclonal antibodies belonging to CD44. The possibility that these higher molecular weight species may represent novel CD44 isoforms containing additional protein sequence was investigated. CD44 cDNA clones were isolated from a plasmid-based expression library prepared from KG1a mRNA. One of the three clones obtained (clone 2.3) was found to encode a CD44 molecule of approximately 130 kD in transfected COS cells. Sequences analysis indicated that the molecule encoded by this cDNA clone, designated CD44R1, was essentially identical to CD44 except for the presence of an additional 132 amino acids inserted into the extracellular domain. This inserted region is rich in serine and threonine residues that may serve as sites of O-linked glycosylation, and contains a potential site of N-linked glycosylation and a potential site of chondroitin sulphate attachment. PCR analysis using primers that flank the inserted region present within CD44R1 identified an additional CD44 isoform, designated CD44R2, that contains only the last 69 amino acids present within the unique region of CD44R1. Peripheral blood mononuclear cells and granulocytes from normal individuals and patients with chronic myelogenous leukemia, polycythemia vera, or acute myelomonocytic leukemia, express both CD44R1 and CD44R2. In contrast, CD44R1 and CD44R2 appear to be differentially expressed in various CD44-positive cell lines. Thus KG1a, and the Epstein-Barr Virus-transformed B cell lines WalkDR4 and Way-1 express both CD44 and the CD44 isoforms CD44R1 and CD44R2, while the myeloid cell lines HL60 and U937 express high levels of CD44, but only very low levels of CD44R1 and CD44R2. The CD44-negative cell lines DHL-4, DHL-10, Jurkat, and K562 are also negative for CD44R1 and CD44R2.
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Han, Yajun, Hesham M. Amin, Bevin Franko, Christine Frantz, Xinzhe Shi, and Raymond Lai. "Loss of SHP1 enhances JAK3/STAT3 signaling and decreases proteosome degradation of JAK3 and NPM-ALK in ALK+ anaplastic large-cell lymphoma." Blood 108, no. 8 (October 15, 2006): 2796–803. http://dx.doi.org/10.1182/blood-2006-04-017434.
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AbstractPrevious studies showed that most cases of ALK+ anaplastic large-cell lymphoma (ALK+ALCL) do not express SHP1, a tyrosine phosphatase and an important negative regulator for cellular signaling pathways such as that of JAK/STAT. To fully assess the biologic significance of loss of SHP1 in ALK+ALCL, we transfected SHP1 plasmids into 2 SHP1-, ALK+ALCL cell lines, Karpas 299 and SU-DHL-1. After 24 hours of transfection, pJAK3 and pSTAT3 were decreased, and these changes correlated with down-regulation of STAT3 downstream targets including cyclin D3, mcl-1, and bcl-2. Expression of SHP1 in these 2 cell lines also resulted in marked decreases in the protein levels of JAK3 and NPM-ALK, and these effects were reversible by proteosome inhibitor MG132. Conversely, when SHP1 expression in SUP-M2 (a SHP1+ ALK+ALCL cell line) was inhibited using siRNA, pSTAT3, pJAK3, JAK3, and NPM-ALK were all up-regulated. Coimmunoprecipitation studies showed that SHP1 was physically associated with JAK3 and NPM-ALK. SHP1 expression in Karpas 299 and SU-DHL-1 led to significant G1 cell cycle arrest but not apoptosis. To conclude, loss of SHP1 contributes to the pathogenesis of ALK+ALCL by 2 mechanisms: (1) it leaves the tyrosine phosphorylation and activation of JAK3/STAT3 unchecked and (2) it decreases proteosome degradation of JAK3 and NPM-ALK.
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Wang, Huaizhu, Mengjie Zhang, Chao Chen, and Baozhen Yao. "Air parcel network design considering pure freighters." SIMULATION 95, no. 9 (November 10, 2017): 797–807. http://dx.doi.org/10.1177/0037549717738336.
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This paper proposes a selection model of candidate nodes and an air parcel network hub location model to estimate the efficiency of the parcel network when adding pure freighters. Using the data on real demands among airports in practice, this article analyzes the hub location problem of the parcel network for DHL Express in the Asia-Pacific region. We find that Beijing, Hong Kong, Singapore, and Mascot are potential choices to be hub airports. The four hubs have parcel production of 10.1, 5.36, 2.75 and 0.84 million tons in the planning horizon, respectively. A sensitivity analysis is conducted to analyze the influence of different numbers of pure freighters on the air parcel network and the results indicate that the number of pure freighters can alleviate parcel pressure.
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Rao, M. S. "Innovative tools and techniques to ensure effective employee engagement." Industrial and Commercial Training 49, no. 3 (March 6, 2017): 127–31. http://dx.doi.org/10.1108/ict-06-2016-0037.
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Purpose The purpose of this paper is to offer innovative tools and techniques to ensure employee engagement. Design/methodology/approach The paper outlines the reasons for employee disengagement and enlightens the advantages of employee engagement for both employees and organizations. It unfolds several research findings on employee engagement and illustrates with the examples of global companies including Cummins, DHL Express, Southwest Airlines, Google, and Virgin. Findings It concludes that employee engagement is a two-way street. Practical implications The tools and techniques adopted by leaders can be applied in any industry and in any size of organization. Social implications The social implications of this research suggest that leaders can ensure employee engagement by following these innovative tools and techniques. Originality/value It implores both employers and employees to take ownership of their roles and responsibilities to achieve organizational excellence and effectiveness.
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Gozgit, Joseph M., Youngchul Song, Scott Wardwell, Sara Nadworny, Yaoyu Ning, and Victor M. Rivera. "Potent Preclinical Activity of Ponatinib in Germinal Center B-Cell-like Diffuse Large B-Cell Lymphoma (GCB-DLBCL) Models." Blood 126, no. 23 (December 3, 2015): 4000. http://dx.doi.org/10.1182/blood.v126.23.4000.4000.
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Abstract Introduction Diffuse large B-cell lymphoma (DLBCL), the most common type of Non-Hodgkin lymphoma (NHL), comprises 2 major molecular subtypes: germinal center B-cell-like (GCB) and activated B cell-like (ABC). Although standard therapy (rituximab+ chemotherapy [R-CHOP]) is effective in most patients (pts), a significant proportion do not achieve durable remissions. Treatment of relapsed and refractory DLBCL pts with targeted therapy, such as the BTK inhibitor ibrutinib, has shown some promise; however, responses are mostly restricted to the ABC subtype. Treatment options for pts with relapsed/refractory GCB, outside of stem cell transplantation, are especially limited. Ponatinib is a potent pan-BCR-ABL inhibitor approved for pts with refractory or T315I+ chronic myeloid leukemia or Ph+ acute lymphoblastic leukemia. Initial characterization of the in vitro kinase activity of ponatinib demonstrated substantial activity against a number of additional oncogenic kinases, including KIT, RET, FLT3, and members of the FGFR, PDGFR, and SRC families. To obtain a broad, unbiased, assessment of the anti-proliferative effects of ponatinib, we screened a panel of 246 human tumor cell lines. Based on the novel finding that a GCB-DLBCL cell line was amongst those inhibited most potently by ponatinib, we conducted studies to further characterize the activity of ponatinib in NHL, and GCB-DLBCL in particular. Results A broad cell-based screen identified a small subset of cell lines (18/246; 7%) whose growth was potently inhibited by ponatinib (GI50<42 nM). A majority of these lines express activated variants of previously validated targets of ponatinib: ABL (N=5, GI50 <0.3 nM), FLT3 (N=1, GI50 1 nM), FGFR2 (N=2, GI50s 5-29 nM), and PDGFRα (N=1, 14 nM). In addition, ponatinib potently inhibited growth of the GCB-DLBCL cell line DoHH2 (GI50 8 nM). The cellular activity of ponatinib was next examined in a larger set of NHL cell lines enriched for the GCB subtype (Table 1). Ponatinib only exhibited modest activity (GI50 46-119 nM) against 2 mantle cell lymphoma (MCL) lines, but potently inhibited growth (GI50≤10 nM) of the one Burkitt's lymphoma (BL) line tested (Daudi). Most notably, ponatinib also potently inhibited growth of 5/9 GCB cell lines. In contrast, none of the GCB lines showed sensitivity to ibrutinib (GI50s >100 nM). Finally, we evaluated the in vivo potency of ponatinib in mice implanted with the GCB cell lines exhibiting the greatest (SU-DHL-4) and weakest (SU-DHL-10) in vitro sensitivity to ponatinib, using dosing regimens previously shown to be active in BCR-ABL models predictive of efficacy in patients. Once-daily oral administration of ponatinib resulted in a dose-dependent inhibition of SU-DHL-4 tumor growth, with 10 mg/kg inducing 78% tumor regression, and 30 mg/kg rapidly inducing complete regression that was maintained in all mice for an additional 2 weeks after ponatinib dosing was stopped. In contrast, ponatinib had much more modest effects on SU-DHL-10 tumors with 30 mg/kg only inhibiting tumor growth by 39%. Conclusion Ponatinib has promising in vitro and in vivo activity against a substantial subset of GCB-DLBCL models tested, with potency similar to that observed in BCR-ABL models. These results provide support for evaluating ponatinib in GCB-DLBCL pts who have failed prior therapy. Studies to further characterize the molecular basis for the activity of ponatinib in NHL are ongoing. Table 1. In vitro drug activity in 12 NHL cell lines Cell line Type Ponatinib GI50 (nM) Ibrutinib GI50 (nM) SU-DHL-4 GCB DLBCL 1.3 313 DoHH2 GCB DLBCL 2.5 114 Pfeiffer GCB DLBCL 6 2,074 SU-DHL-6 GCB DLBCL 9.8 1,041 WSU-NHL GCB DLBCL 10 1,672 Farage GCB DLBCL 51 1,409 U-2932 GCB DLBCL 79 >10,000 RL GCB DLBCL 212 6,939 SU-DHL-10 GCB DLBCL 238 2,827 Daudi BL 2.9 4,319 Mino MCL 46 >10,000 Jeko-1 MCL 119 4,781 GI50: the concentration that causes 50% growth inhibition. Disclosures Gozgit: ARIAD Pharmaceuticals Inc.: Employment, Other: Full-time Employee & Shareholder (self-managed). Song:ARIAD Pharmaceuticals Inc.: Employment, Other: Full-time Employee & Shareholder (self-managed). Wardwell:ARIAD Pharmaceuticals Inc.: Employment, Other: Full-time Employee & Shareholder (self-managed). Nadworny:ARIAD Pharmaceuticals Inc.: Employment, Other: Full-time Employee & Shareholder (self-managed). Ning:ARIAD Pharmaceuticals Inc.: Employment, Other: Full-time Employee & Shareholder (self-managed). Rivera:ARIAD Pharmaceuticals Inc.: Employment, Other: Full-time Employee & Shareholder (self-managed).
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Ysebaert, Loic, Laurence Lamant, Estelle Espinos, Sylvie Giuriato, Georges Delsol, and Guy Laurent. "ALK Activates a Non-Canonical Beta-catenin Pathway in Anaplastic Large Cell Lymphoma." Blood 112, no. 11 (November 16, 2008): 5265. http://dx.doi.org/10.1182/blood.v112.11.5265.5265.
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Abstract Molecular profiling of Anaplastic Lymphoma Kinase (ALK)-positive versus negative cell lines and patients’ samples has unravelled critical roles for transcriptional factors (TF) (C/EBPbeta, Bcl-6) in Anaplastic Large Cell Lymphomas (ALCL), suggesting that targeting these TF with ALK tyrosine kinase inhibitors should yield better results (Lamant L et al., Blood 2008). In this latter work of our group, beta-catenin and its transcriptional partner T Cell Factor 4 (TCF4) were also found upregulated at a transcriptional level in ALK+ vs ALK− patients’ samples and cell lines. We sought to determine the relevance of such a finding. At the protein level, ALK+ cell lines (SU-DHL-1, Karpas299 and COST) and ALK− cell line (FEPD) express detectable beta-catenin. TCF-4 is also expressed at similar levels among cell lines, whatever ALK status. Since beta-catenin is mainly regulated through post-transcriptional mechanism, we assessed its phosphorylation status. ALK+ (SU-DHL-1 and Karpas299), but not ALK− cell lines displayed increased tyrosine phosphorylation at both Tyr142 and Tyr654 residues, as well as association between beta-catenin and ALK (in immuno-precipitation assays). Moreover, both total beta-catenin and phosphoTyr-beta-catenin were found associated with TCF4, and therefore transcriptionally active. Using the tetracycline system to allow conditional expression of NPM-ALK in MEF cell line (murine embryonic fibroblasts), we found that beta-catenin phosphorylation became barely detectable upon loss of NPM-ALK expression. Based on these findings, we investigated the functional consequences of the disruption of beta-catenin/TCF4 complexes using small molecules such as PKF115–584 and CGP049090 (a generous gift from Novartis). Interestingly, both compounds induced dissociation of beta-catenin/TCF4 complexes at 0.5μM for 24h, and also induced apoptosis in SU-DHL-1 cells. But, since compounds could not dissociate beta-catenin/ALK complexes, they should be combined to ALK inhibitors to fully exert their anti-lymphoma effects. Moreover, the pool of beta-catenin linked to Glycogene Synthase Kinase 3beta (regulated by external Wnt-dependant signals) is neither affected by small compounds, indicating a specific beta-catenin/TCF4 disruption with these drugs. To conclude, this study shows that, in ALCL, ALK activates a Wnt-independent pathway, which appears to be critical for cell survival. This study offers a rationale for investigating the potential of molecules designed to interfere with beta-catenin/TCF/LEF proteins and currently evaluated in colon carcinomas, alone or in combination with ALK tyrosine kinase inhibitors.
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Baxter, Glenn, and Panarat Srisaeng. "Cooperating to Compete in the Global Air Cargo Industry: The Case of the DHL Express and Lufthansa Cargo A.G. Joint Venture Airline ‘AeroLogic’." Infrastructures 3, no. 1 (March 16, 2018): 7. http://dx.doi.org/10.3390/infrastructures3010007.
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Buttiglieri, Stefano, Carmelo Carlo-Stella, Tiziana Spatola, Roberta Pulito, Luigi Naldini, Andrea Anichini, Michele Magni, Arianna Giacomini, Corrado Tarella, and Alessandro M. Gianni. "Potent In Vivo Anti-Tumor Activity Of Extracellular Vesicles Isolated From Genetically Engineered Primary Mesenchymal Stromal Cells Expressing The Trans-Membrane TNF-Related Apoptosis-Inducing Ligand (TRAIL)." Blood 122, no. 21 (November 15, 2013): 1658. http://dx.doi.org/10.1182/blood.v122.21.1658.1658.
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Abstract Introduction TNF-related apoptosis-inducing ligand (TRAIL) is a protein functioning as a ligand that induces the process of cell death. TRAIL has been shown to kill in vitro a wide variety of tumor cells with minimal effects on normal cells. Despite its in vitro activity, recombinant soluble TRAIL has so far shown limited efficacy in vivo. In contrast, recent reports have shown that significant apoptosis can be observed both in vitro and in vivo when TRAIL is expressed on the cell membrane (mTRAIL). A further innovation might be the delivery of bioactive proapoptotic TRAIL through its expression by extracellular vescicles (EVs), the nanovesicular organelles secreted by cells. In fact, EVs are viewed as an effective tool for intercellular cross-talk and receptor discharge. The trans-membrane expression of TRAIL ligand within the double layer exosomal membrane may induce a more potent death signal when compared with the soluble molecule. Material and Methods Mesenchymal Stromal Cells (MSC) from bone marrow were cultured in vitro and used for EVs production. Cultured MSC in 75 cm2 flasks, at 80% confluence were infected with a lentivector encoding TRAIL, maintained in culture, and cell-supernatants repeatedly collected over several days, ultracentrifugated, with EVs-containing pellet harvested in PBS. EVs were produced also from uninfected MSC as control (EVs-CTRL). EVs were characterized by flow cytometry for expression of MSC markers and mTRAIL, EV size was evaluated by NanoSight technology. Total protein concentration was used to quantify EVs, Western Blot analysis was performed to characterize membrane-bound TRAIL. In vitro analysis was performed on SU-DHL-4 (human B cell lymphoma) and MEL-1300 (human melanoma) cell lines, exposed for 24 hours to 20-100 μg/ml EVs-TRAIL or EVs-CTRL. Annexin/propidium iodide assay was used to quantify apoptotic/necrotic cells. For the in vivo assessments, SU-DHL-4 and MEL-1300 cells were transduced with Luc-Lentiviral particles to obtain Luciferase positive cell lines. These cells were used to engraft NOD scid gamma (NSG) mice (2x106 SU-DHL-4 and 3x105 MEL-1300 cells for each subcutaneous injection point). To visualize tumor cells, mice were injected intraperitoneum with luciferin and analyzed with the Xenogen system. Mice bearing subcutaneous tumor nodules received single intravenous injections of 100, 200, 300 µg or multiple (x 3) 200 µg injections of either EVs-TRAIL or EVs-CTRL. Results FACS analysis showed strong TRAIL expression on EVs from TRAIL-infected MSC compared to EVs-CTRL, with a high proportion of positive particles (median 85%, range 78-93). In addition, EVs-TRAIL displayed MSC membrane markers, i.e. CD 105, CD 90, CD73 and CXCR4. Western Blot analysis under non-reducing conditions showed the presence of TRAIL ligand, with strong prevalence of dimeric TRAIL isoform (barely detectable the trimeric isoform, undetectable monomeric isoforms). NanoSight analysis revealed that EVs had a variable size, up to approximately 400 nm in diameter, with a predominant peak at 273 nm. A strong and dose-dependent cytotoxic effect was observed on SU-DHL-4 cells exposed to EVs-TRAIL (annexin/PI+ve cells: up to 87% for 100 μg/ml EVs-TRAIL), compared to EVs-CTRL exposure (15% Annexin/PI+ve cells for 100 μg/ml EVs-TRAIL). A similar, albeit less pronounced in vitro cytotoxic effect of EVs-TRAIL was observed on the melanoma MEL-1300 cell line. The anti-tumor effect was remarkably strong when EVs-TRAIL were injected in vivo in mice bearing either SU-DHL-4 or MEL-1300 nodules. A marked reduction of the tumor luminescence from 1.2x1010 photon/sec to <108 photon/sec was observed at seven days since a single EVs-TRAIL injection at 200 and 300 μg. Multiple administrations of 200 μg EVs-TRAIL induced the strongest luminescence reduction, as observed in MEL-1300 bearing NSG mice. Histological examination of nodules from EVs treated mice showed necrosis areas along with extensive intra-tumor vascular disruption. Conclusion EVs isolated from genetically engineered TRAIL-expressing MSC: i. do express mTRAIL; ii. display potent antitumor activity, inducing extensive apoptosis/necrosis both in vitro and in vivo in animal models bearing lymphoma and melanoma nodules. Thus, EVs-TRAIL may represent a promising strategy for delivering pro-apoptotic signals to tumor cells. Moreover, the Results could pave the way to the use of EVs for therapeutic purposes. Disclosures: No relevant conflicts of interest to declare.
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Schuster, Stephen J., Jakub Svoboda, Sunita Dwivedy Nasta, Elise A. Chong, Nicole Winchell, Daniel J. Landsburg, David L. Porter, et al. "Treatment with Chimeric Antigen Receptor Modified T Cells Directed Against CD19 (CTL019) Results in Durable Remissions in Patients with Relapsed or Refractory Diffuse Large B Cell Lymphomas of Germinal Center and Non-Germinal Center Origin, "Double Hit" Diffuse Large B Cell Lymphomas, and Transformed Follicular to Diffuse Large B Cell Lymphomas." Blood 128, no. 22 (December 2, 2016): 3026. http://dx.doi.org/10.1182/blood.v128.22.3026.3026.
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Abstract BACKGROUND: The cell of origin (COO) of diffuse large B cell lymphoma (DLBCL), germinal center (GC) or non-germinal center (NGC), may have prognostic significance for treatment outcome in first-line and relapsed settings (Lenz et al NEJM 2008; Thieblemont et al JCO 2011). "Double hit" DLBCL (DHL), defined by chromosomal breakpoints affecting the MYC/8q24 locus and BCL2/18q21 and/or BCL6/3q27 loci and arising either from transformation of follicular lymphoma (tFL) or de novo, has no standard effective therapy in the relapsed setting. Since new therapies are needed for poor prognostic groups of relapsed DLBCL patients (pts), we examined the efficacy of treatment with autologous T cells genetically modified to express a chimeric antigen receptor consisting of an external anti-CD19 single chain murine antibody domain with CD3ζ and 4-1BB signaling domains (CTL019 cells) in pts with relapsed/refractory GC and NGC DLBCL, DHL, and tFL as part of an ongoing phase IIa clinical trial (NCT02030834). METHODS: Eligible pts had CD19+ DLBCL with measurable residual disease after primary and salvage therapies, were not eligible for autologous stem cell transplant (ASCT) or had relapsed/residual disease after ASCT, and had responsive or stable disease with most recent therapy. COO of DLBCL was defined immunohistochemically using the Hans algorithm (Hans et al Blood 2004) and included the DLBCL subtypes DHL and tFL that met this definition. Fluorescence in-situ hybridization was performed on diagnostic tissue using Vysis MYC (8q24), BCL2 (18q21) and BCL6 (3q27) break apart probes to determine DHL. DHL was defined by a MYC locus chromosomal translocation with a second translocation involving either BCL2 or BCL6. After steady state apheresis to collect peripheral blood leukocytes, pts received lymphodepleting chemotherapy based on clinical features and past therapies. 1 to 4 days after chemotherapy, pts received a single dose of CTL019 cells. Following CTL019, pts received no further therapy for DLBCL. Initial tumor response assessment was performed 3 months (mo) after CTL019 using IWG criteria. Enrollment started in Feb 2014; data are reported through 24 Jul 2016. RESULTS: 13 pts with DLBCL are enrolled and evaluable for response (7 pts GC, 5 pts NGC, 1 undetermined). The median age is 59 years (range: 25-77), male:female ratio 10:3, median number of prior therapies 5 (range: 2-8), and number of pts with prior transplant 7 (54%). At enrollment, Ann Arbor stages were: Stage IV 8 pts (61%), Stage III 1 pt (8%), and Stage II 3 pts (23%) Stage IE 1 pt (8%); 3 pts (23%) had bone marrow involvement. LDH was increased in 8 pts (62%). ECOG PS was 0 in 4 pts (31%) and 1 in 9 pts (69%).Lymphodepleting chemotherapy regimens were bendamustine (90 mg/m2 x 2; 1 pt), cyclophosphamide (1 gm/m2; 2 pts), radiation-cyclophosphamide (4,000cGy-750 mg/m2; 1 pt), modified-EPOCH (3 pts), and hyper-fractionated cyclophosphamide (300 mg/m2 q12 hours x 6; 6 pts). 12 pts received 5.00E+08 (5.10 - 6.75E+06 cells/kg) CTL019 cells; 1 pt received 1.93E+08 (3.10E+06 cells/kg).Median peak CTL019 cell expansion in blood occurred 7 days after infusion (range: 2-28 days); there is no difference in peak expansion between responders and non-responders or pts with GC or NGC DLBCL.Cytokine release syndrome occurred in 9 pts (8 grade 2; 1 grade 3) and did not predict response. Transient neurotoxicity included delirium in 2/13 pts (1 grade 2; 1 grade 3) and cognitive disturbance in 1/13 pts (1 grade 1). At 3 mo post CTL019, overall response rate (ORR) is 52% (7/13 pts); ORR at 3 mo for GC 71% (5/7 pts) and NGC 40% (2/5 pts). Complete response rate (CR) at 3 mo is 38% (5/13 pts); CR at 3 mo for GC 43% (3/7 pts) and NGC 40% (2/5 pts). Best response for all pts is CR in 6 of 13 pts (46%); CR for GC 57% (4/7 pts) and NGC 40% (2/5 pts). 3 of 7 pts with GC DLBCL had tFL and all 3 achieved CR; 2 of 7 pts with GC DLBCL had DHL and both achieved CR. To date, no pt achieving CR has relapsed. Median progression-free survival is 5.8 mo for all pts, 3.0 mo for NGC pts, and not reached for GC pts (57.1% [95%CI: 17.2%-83.7%] progression-free at median follow-up 21.9 mo). At median follow-up 23.3 mo for responding pts, 85.7% [95%CI: 33.7-97.9%] maintain response. CONCLUSIONS: A single treatment with CTL019 cells can achieve durable remissions in pts with relapsed/refractory GC and NGC DLBCL, DHL and transformed FL. Figure 1 Figure 1. Figure 2 Figure 2. Disclosures Schuster: Nordic Nanovector: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Celgene: Consultancy, Honoraria, Research Funding; Pharmacyclics: Consultancy, Research Funding; Genentech: Consultancy, Honoraria; Merck: Research Funding; Janssen Research & Development: Research Funding; Hoffman-LaRoche: Research Funding; Gilead: Research Funding; Novartis: Consultancy, Honoraria, Research Funding. Svoboda:Pharmacyclics: Research Funding; Celgene: Research Funding; Seattle Genetics: Research Funding. Nasta:Millennium Pharmaceuticals: Research Funding. Porter:Genentech/Roche: Employment; Genentech/Roche: Equity Ownership; Incyte: Honoraria; Novartis: Consultancy; Novartis: Research Funding; University of Pennsylvania: Patents & Royalties; Immunovative Therapies: Other: Travel, Accommodations, Expenses. Mato:ProNAi: Research Funding; TG Therapeutics: Research Funding; Acerta Pharma: Research Funding; Theradex: Research Funding; Gilead Sciences: Research Funding; Abbvie: Research Funding; TG Therapeutics: Consultancy; Pharmacyclics: Consultancy; Gilead Sciences: Consultancy; Abbvie: Consultancy. Wasik:Gilead Sciences: Equity Ownership; Seattle Genetics: Honoraria; Novartis: Research Funding; University of Pennsylvania: Patents & Royalties: NPM-ALK as an omncogene; University of Pennsylvania: Patents & Royalties: CAR T-cells; Gilead Sciences: Research Funding; Pharmacyclics: Research Funding. Lacey:Novartis: Research Funding. Melenhorst:Novartis: Patents & Royalties: Novartis, Research Funding. Chew:Novartis: Research Funding. Hasskarl:Novartis: Employment. Marcucci:Novartis: Research Funding. Levine:GE Healthcare Bio-Sciences: Consultancy; Novartis: Patents & Royalties, Research Funding. June:Pfizer: Honoraria; Johnson & Johnson: Research Funding; University of Pennsylvania: Patents & Royalties; Tmunity: Equity Ownership, Other: Founder, stockholder ; Novartis: Honoraria, Patents & Royalties: Immunology, Research Funding; Celldex: Consultancy, Equity Ownership; Immune Design: Consultancy, Equity Ownership.
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Staber, Philipp B., Werner Linkesch, Silvia Schauer, Gerit Moser, Marshall E. Kadin, Willi Dirks, and Gerald Hoefler. "Genes Regulated by NPM-ALK Fusion Kinase Play a Key Role in the Activation of AP-1 Transcription Factors." Blood 104, no. 11 (November 16, 2004): 245. http://dx.doi.org/10.1182/blood.v104.11.245.245.
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Abstract Background: About half of nodal anaplastic large cell lymphomas (ALCL) express the nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) fusion protein, which is the product of a t(2;5)(p23;q35) chromosomal translocation. Expression of this protein has been shown to result in neoplastic change. Combining suppression subtractive hybridization (SSH) and cDNA microarray analysis we aimed at elucidating the consequences of NPM-ALK expression. Methods: SSH cDNA libraries were constructed using mRNA from human embryonic kidney (293) cells transfected with active or kinase-dead NPM-ALK constructs, as well as pools of NPM-ALK positive and negative ALCL cell lines. The resulting cDNA clones and genes relevant for cancer pathogenesis were spotted, generating specific cDNA microarrays comprising 4992 genes. mRNA expression patterns were analyzed in individual cell lines. Real time quantitative RT-PCR of 20 selected genes validated the mRNA expression data of the microarrays. Results: Expression of a set of 102 genes distinguishes NPM-ALK-negative (FE-PD, MAC-2A) from NPM-ALK-positive ALCL cell lines (SU-DHL-1, JB-6, SUP-M2, SR-786, DEL and Karpas 299). The majority are involved in regulation of cell cycle and apoptosis. 38 of these genes also discriminate 293 cells with respect to their NPM-ALK expression. Interestingly, AP-1 target genes, such as GM-CSFRA, GM-CSFRB, ARF5, FAS, FASL, and BCL3, are increased in NPM-ALK expressing cell lines. Electrophoretic mobility shift assay (EMSA) verifies NPM-ALK dependent AP-1 DNA binding activity. Conclusion: This study reveals genes specifically regulated by NPM-ALK lymphoma kinase. Further, we demonstrate that AP-1 activation is a critical target of NPM-ALK signalling.
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Wilcox, Ryan A., Andrew L. Feldman, Steven C. Ziesmer, Anne J. Novak, Thomas E. Witzig, and Stephen M. Ansell. "GATA-3 Promotes IL-10 Production and Is An Adverse Prognostic Factor in Peripheral T-Cell Lymphoma Unspecified (PTCL-U)." Blood 114, no. 22 (November 20, 2009): 3929. http://dx.doi.org/10.1182/blood.v114.22.3929.3929.
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Abstract Abstract 3929 Poster Board III-865 In addition to providing prognostic information, gene expression profiling studies have helped to elucidate the putative cell of origin (e.g. activated or germinal center B cell) in diffuse large B-cell lymphoma. With the exception of angioimmunoblastic T-cell lymphomas, which are thought to derive from follicular helper T cells, little is known about the cell of origin for the most common peripheral T-cell lymphoma (PTCL), PTCL-unspecified (PTCL-U). Following appropriate antigenic stimulation, naïve CD4+ T cells may terminally differentiate into effector cells, defined by the production of specific cytokines, including IL-4/IL-10 producing Th2 cells. We have previously demonstrated that IL-10, produced by malignant T cells, is systemically elevated in the serum of patients with PTCL and contributes to the suppression of host anti-tumor immunity. Murine studies have demonstrated that the transcription factor GATA-3 is both critically important for IL-10 transcription and is required for the differentiation of Th2 cells. Therefore, we hypothesized that a subset of PTCL-U may be Th2-derived and express GATA-3. To examine this possibility, PTCL-U cases with adequate paraffin-embedded tissue were identified from a single institution (n=46) and immunohistochemically stained for GATA-3 expression. More than 10% of tumor cells expressed GATA-3 in 17 (37%) of the 46 cases examined. Follow up data sufficient to determine overall survival was available for 43 of these patients, in whom the expression of GATA-3 was associated with a poorer overall survival. Patients with GATA-3 negative tumors (n=27) had a median overall survival (OS) of 3.8 years, whereas patients with GATA-3 positive tumors (n=16) had a median OS of 0.7 years (p=.002). Among low-intermediate risk patients, as defined by the International Prognostic Index (IPI), GATA-3 expression was associated with inferior median overall survival (<12 months, p<.001). IL-10 production may explain the adverse impact of GATA-3 expression on OS in PTCL-U, as GATA-3 plays a central role in IL-10 transcription in normal T cells. To determine whether it may have a similar role in malignant T cells, we identified a PTCL cells line (SU-DHL-1) that highly expressed GATA-3 by intracellular flow cytometry. We next transfected these cells with a negative control or GATA-3 specific siRNA and demonstrated that transfection with GATA-3 siRNA significantly decreased GATA-3 expression in these cells. Following knockdown of GATA-3 expression, we measured both cell proliferation by 3H-TdR incorporation and IL-10 production by ELISA. No significant difference in cell proliferation was observed (n=4, p=0.2), whereas an approximately 50% reduction in IL-10 production was observed (n=3, p=.008) following inhibition of GATA-3 expression. Therefore, GATA-3 expression in malignant T cells may promote the production of IL-10, thus impairing host anti-tumor immunity. Immunomodulatory drugs (IMiDs), like lenalidomide, may downregulate GATA-3 expression in normal T cells. Therefore, we sought to determine whether lenalidomide may inhibit IL-10 production in normal and malignant T cells. T cells obtained from normal donors were activated with anti-CD3 and treated with either DMSO or lenalidomide (10 μM). Intracellular staining for both IL-10 and IL-2 was performed 3 days later. In DMSO control treated cells, 1.0% of cells expressed IL-10 and 12% expressed IL-2. In contrast, in lenalidomide treated cells, only 0.3% of cells expressed IL-10 and 17% expressed IL-2 (representative of at least 4 independently performed experiments). Similar results were obtained using T cells from a patient with Sezary Syndrome. We next treated SU-DHL-1 cells with DMSO or lenalidomide (0.1, 1.0 and 10 μM) and measured both cell proliferation by 3H-TdR incorporation and IL-10 production by ELISA. Significant inhibition of SU-DHL-1 proliferation was only observed when cells were treated with 10 μM lenalidomide. In contrast, significant inhibition of IL-10 production was observed in the presence of 1 μM and 10 μM lenalidomide (p<.01). Collectively, the data presented demonstrates that GATA-3 promotes the production of IL-10 by malignant T cells and is an adverse prognostic factor in PTCL-U. Given its immunomodulatory properties, including inhibition of IL-10 production, lenalidomide represents a promising therapeutic agent in PTCL-U. Disclosures: No relevant conflicts of interest to declare.
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Vogiatzi, Fotini, Julia Heymann, Thies Rösner, Lennart Lenk, Gunnar Cario, Martin Schrappe, Beat Bornhauser, et al. "Venetoclax Enhances the Efficacy of Therapeutic Antibodies in B-Cell Malignancies." Blood 132, Supplement 1 (November 29, 2018): 4177. http://dx.doi.org/10.1182/blood-2018-99-114677.
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Abstract The application of antibodies is a promising option in the treatment of B-cell malignancies, including acute lymphoblastic leukemia (ALL) and B-cell Non-Hodgkin lymphoma (B-NHL). Although patient outcomes have improved by applying combinations of chemotherapy and antibodies, certain patients characterized by a high expression of anti-apoptotic Bcl-2 have a poor prognosis. These include adult B-NHL patients with "double-hit lymphomas" (DHLs) and pediatric ALL patients harboring a t(17;19) translocation. Furthermore, a substantial number of Burkitt´s lymphoma (BL) patients also express Bcl-2 even though the impact of this finding on prognosis is yet unclear. Here, we examine the role of low doses of the Bcl-2 inhibitor venetoclax (VTX, 1nM) on the efficacy of the therapeutic antibodies rituximab (CD20), daratumumab (CD38) and CD19-DE (a variant of the CD19 antibody MOR208 engineered for improved effector cell binding). Natural killer (NK)-cell mediated antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent cell-mediated phagocytosis (ADCP) were evaluated. The CD20-expressing DHL cell line Carnaval and patient-derived xenograft (PDX) cells from two BL patients were used as target cells for rituximab, the CD20-negative/CD38-positive DHL cell line Will-2 was used with daratumumab and three PDX samples from t(17;19) positive ALL patients were used with CD19-DE. For the assessment of ADCP, human monocyte-derived macrophages were incubated with labelled target cells and microscopy assays were performed. NK-cell mediated ADCC was not enhanced by VTX in any of our models. However, 17-37% increases in ADCP by macrophages were detected when Carnaval cells were subjected to combinations of VTX/rituximab and when Will-2 cells were treated with VTX/daratumumab as compared to VTX or antibody alone (p=0.0318/p=0.0185 and p=0.0012/p=0.0068, respectively, Figure A). When BL PDX cells were subjected to ADCP assays with VTX/rituximab, mean phagocytosis levels were also enhanced by 26.0% and 21.0% in the combination treatment group as compared to VTX (p=0.0283) and rituximab alone (p=0.0282; Figure A). ADCP assays with t(17;19) positive ALL-PDX cells and CD19-DE confirmed these results as phagocytosis was increased to similar extents in the combination group as compared to VTX (p=0.0017) or CD19-DE alone (p=0.0323) (Figure A/B). In order to exclude that our observations were due to an enhancement of apoptosis in target cells only, we measured cleaved caspase-3 with VTX, antibodies alone or the combination of both. Cleaved caspase-3 levels were equal in all groups suggesting that the addition of VTX resulted in an apoptosis-independent activation of macrophages. In order to minimize heterogeneity in ADCP assays, phagocytosis was next examined using expanded macrophages from NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice for our assays. Regardless of target cells and antibody, the combination of antibody treatment with VTX resulted in enhanced phagocytosis by murine macrophages, confirming our results. The effects of VTX on the efficacy of rituximab were finally examined in vivo. Carnaval cells were injected intravenously into NSG mice and animals treated with VTX (100 mg/kg 5 days/week by oral gavage), rituximab alone (1 mg/kg once weekly intraperitoneally) or the combination of both (n=6/group). Mice were sacrificed when mice showed clinical lymphoma or leukemia engraftment and survival differences were assessed using Kaplan-Meier log-rank statistics. Compared to control, mice treated with VTX displayed a slight survival advantage, which was more marked in mice treated with rituximab (p=0.0020/p=0.0004, respectively, Figure C), suggesting a better efficacy of rituximab than VTX as monotherapy. Most importantly, mice treated with the combination VTX/rituximab showed significantly superior survival as compared to either VTX or rituximab alone (p=0.0023/p=0.0268, respectively, Figure C), suggesting additive effects in vivo. Altogether, we show that VTX enhances the efficacy of therapeutic antibodies in models of B-cell malignancies including PDX samples. The mechanism is most likely dependent on distinct influences of VTX on macrophage activation, e. g. by myeloid immune checkpoints. Our in vivo data suggest that this combination strategy may become a promising therapeutic option for the treatment of Bcl-2 expressing B-cell malignancies in the future. Figure. Figure. Disclosures Bourquin: Amgen: Other: Travel Support. Valerius:Affimed: Research Funding. Peipp:Affimed: Research Funding.
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Berlier, Willy, Karine Aguera, Anne-Marie Chevrier, Fanny Gallix, Alexandra Traverse-Glehen, Gilles Salles, and Yann Godfrin. "Asparagine Synthetase Expression and L-Asparaginase Sensitivity in Aggressive Lymphomas." Blood 124, no. 21 (December 6, 2014): 5494. http://dx.doi.org/10.1182/blood.v124.21.5494.5494.
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Abstract L-asparaginase (L-ASPA) displays a strong clinical benefit in the treatment of acute lymphoblastic leukemia (ALL), where it is included in most of current chemotherapy regimen. L-ASPA depletes plasmatic asparagine (ASN), an amino acid essential for the proliferation of leukemic cells. Since these cells are deficient in asparagine synthetase (ASNS), they rely on external (plasmatic) source of ASN and can be starved to death by L-ASP treatment. Several studies evidenced the potential of ASN depletion to treat lymphomas. Indeed, many animal and human lymphoma cell lines have been shown to be sensitive to L-ASPA in vitro. In veterinary medicine, L-ASPA is routinely administered to treat effectively both feline and canine lymphomas (Wypig et al., 2013). L-ASPA regained attention in the treatment of human lymphomas since its adjunction in current chemotherapy regimens significantly improved the outcome of patients with NK/T cell lymphoma (Zou et al., 2014). Some studies also evidenced its benefit in combined chemo or monotherapy for the treatment of B-cell and T-cell lymphomas (Sun et al., 2006; Takahashi et al., 2010). In this study, we assessed the in vitro sensitivity to L-ASPA of 6 lymphoma cell lines and we analyzed ASNS expression in biopsies from 166 cases of lymphomas (130 B-cell lymphomas and 17 T-cell lymphomas). Sensitivity to L-ASPA (expressed as an IC50) was assessed in vitro by measuring the cell viability in the presence of various concentrations of E.coliL-ASPA. ASNS expression in biopsies (TMA, USBiomax, Rockville, MD) was assessed with a validated immunohistochemistry (IHC) method attributing a score to each tumor based on ASNS labeling intensity from 0 (no expression) to 3 (strong expression). Tumors expressing no/low ASNS (scores 0 and 1) were considered potentially sensitive to asparagine depletion. As shown in the following table, all cell lines were proved to be sensitive to L-ASPA. Their in vitrosensitivity exceeded cell lines MOLT-4 (ALL) and HL-60 (AML). Table 1Cell lineSensitivity to L-ASPA (IC50 in IU/mL)HuT-78 (Peripheral T-cell lymphoma,PTCL)0.11 ± 0.02Toledo (Diffuse large B-cell lymphoma, DLBCL)0.19 ± 0.03SU-DHL-8(Diffuse large B-cell lymphoma, DLBCL)0.10 ± 0.04SU-DHL-10(Diffuse large B-cell lymphoma, DLBCL)0.10 ± 0.01REC-1 (Mantle cell lymphoma, MCL)0.15 ± 0.03KHYG-1 (NK/T-cell lymphoma)0.16 ± 0.06MOLT-4 (acute lymphoid leukemia, ALL)0.19 ± 0.07HL-60 (acute myeloid leukemia, AML)0.23 ± 0.02 As shown in the following table, ASNS expression was null/low in 85% in the entire population of patients with B-cell lymphomas. Considering DLBCL, 63% of patients displayed no ASNS expression at all. ASNS expression was also null/low in 88% of patients with T-cell lymphomas (n=17). Table 2ASNS expression (IHC score)Type of lymphoma(% of cases)DLBCL (n=110)Others BCL (n=20)PTCL (n=3)Others TCL (n=14)MCL(n=3)Hodgkin (n=16)Negative (0)62,770,00,057,133,343,8Low positive (1)21,825,066,635,766,656,3Positive (2)7,35,033,37,10,00,0Highly positive (3)8,20,00,00,00,00,0 Globally, these results suggest that L-ASPA is potentially effective for the treatment of several lymphomas. Indeed, B-cell as well as T-cell lymphoma cell lines are sensitive to L-ASP in vitroand the majority of lymphoma tissues express no/low ASNS. Based on our results on ASNS expression in lymphoma biopsies, L-ASPA therapy may be beneficial for up to 85% of patients with DLBCL. Up to 90% of patients with other B-cell lymphomas or T-cell lymphomas may be sensitive to L-ASPA treatment as well. However, L-ASPA has only been used scarcely in the treatment of lymphomas despite promising clinical responses. Its well known serious side-effects (hypersensitivity, coagulation disorders, pancreatitis, and liver failure) render its use hazardous, particularly in older or frail patients. Therefore, the development of a new formulation of L-ASPA with safer profile has to be considered in order to allow the clinical development of L-ASPA in the treatment of aggressive lymphomas. Disclosures Berlier: ERYTECH: Employment, Equity Ownership. Aguera:ERYTECH: Employment. Chevrier:ERYTECH: Employment. Gallix:ERYTECH: Employment. Godfrin:ERYTECH Pharma: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees.
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Park, Peter U., Véronique Blanc, Jutta Deckert, Pascale Lejeune, Laura M. Bartle, Anna Skaletskaya, Michele F. Mayo, et al. "SAR650984: A Potent Anti-CD38 Therapeutic Antibody with Three Mechanisms of Action (Apoptosis, ADCC, CDC) for Hematological Malignancies." Blood 112, no. 11 (November 16, 2008): 2756. http://dx.doi.org/10.1182/blood.v112.11.2756.2756.
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Abstract CD38 is a promising target for antibody therapeutics for the treatment of various hematological malignancies, including multiple myeloma as well as non-Hodgkin’s lymphoma, chronic lymphocytic leukemia, acute lymphocytic leukemia, acute myeloid leukemia, and chronic myeloid leukemia. CD38 is a type II transmembrane glycoprotein with ectozyme activity that has been implicated in Ca2+ mobilization. CD38 expression correlates with the poor disease prognosis in some hematological malignancies. It has been proposed that rituximab, a well-established anti-CD20 antibody for the treatment of non-Hodgkin’s lymphoma, works by several mechanisms, including antibodydependent cell-mediated cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and apoptosis. A panel of murine anti-CD38 antibodies was first screened for their ability to induce apoptosis, as measured by Annexin V staining, in various lymphoma, leukemia, and multiple myeloma cell lines. Chimeric human IgG1 versions of the selected murine antibodies with potent apopototic activity were then produced and screened for ADCC and CDC activities. Among the screened antibodies, a chimeric version of SAR650984, a humanized anti-CD38 antibody, had the best overall activities. SAR650984 induced potent apoptosis in Daudi, Raji, Ramos, and SU-DHL-8 lymphoma cells, as well as MOLP-8 multiple myeloma (MM) cells and DND-41 acute T lymphocytic leukemia (T-ALL) cells, which like most, if not, all MM and T-ALL cells, express CD38, but not CD20. SAR650984 also induced potent ADCC and CDC in these tumor cells. When the activities of SAR650984 and rituximab were directly compared in Daudi and Raji lymphoma cells that express similar levels of CD38 and CD20, SAR650984 had similar ADCC and CDC activities as rituximab, and, in addition, SAR650984 induced a greater percentage of lymphoma cells to undergo apoptosis than did rituximab. SAR650984 and rituximab showed similar efficacy in a SCID disseminated survival model with Daudi lymphoma cells. Groups of 10 mice were intravenously injected with 5 × 106 Daudi cells on day 0 and then treated with 40 mg/kg SAR650984, 40 mg/kg rituximab, or PBS control on days 7, 11, 14, 18, 21 and 25. The median survival (range) was 28 days (26–30 days) for the PBS-treated group, 47 days (40–51 days) for the SAR650984-treated group and 42.5 days (40–51 days) for the rituximab-treated group. SAR650984 also showed strong anti-tumor activity in a subcutaneous xenograft SCID model with CD38+ CD20− NCI-H929 multiple myeloma cells. Groups of 10 mice were subcutaneously injected with 106 NCI-H929 cells on day 0, and then treated with 40 mg/kg SAR650984, 40 mg/kg rituximab (non-binding IgG1 control), or PBS control twice weekly for three weeks starting on day 6 when the tumors were palpable. A mean tumor volume of 1000 mm3 was reached by PBS-treated group on day 89 and rituximab-treated group on day 84 (70–80% tumor take rate for NCI-H929). SAR650984 treatment completely prevented the tumor growth in all 10 mice (tumor free at the end of study on day 128). In summary, the humanized anti-CD38 antibody, SAR650984, has potent ADCC, CDC, and apoptotic activities in vitro and anti-tumor activity in vivo. SAR650984 is a promising therapeutic antibody candidate for various hematological malignancies, especially in diseases such as multiple myeloma, where rituximab is inactive.
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Leventaki, Vasiliki, Elias Drakos, Megan Lim, Kojo S. Elenitoba-Johnson, Francois-Xavier Claret, L. Jeffrey Medeiros, and George Z. Rassidakis. "NPM-ALK Promotes Cell Cycle Progression through Activation of JNK and c-Jun in Anaplastic Large Cell Lymphoma." Blood 108, no. 11 (November 1, 2006): 2057. http://dx.doi.org/10.1182/blood.v108.11.2057.2057.
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Abstract Anaplastic large cell lymphoma (ALCL) frequently carries the t(2;5)(p23;q35) resulting in aberrant expression of nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) chimeric protein. NPM-ALK mediates its oncogenic effects through phosphorylation of a number of proteins involved in known signal transduction pathways including PLC, PI3K-AKT and JAK-STAT. ALK+ ALCL cells also are known to overexpress c-Jun, a member of the activator protein-1 (AP-1) transcription factor family that controls cell proliferation, differentiation, growth and apoptosis. Phosphorylation of c-Jun at serine 73 and serine 63 residues substantially increases AP-1 transcriptional activity and the levels of c-Jun protein through an autoregulatory positive feedback loop. In this study, we hypothesized that NPM-ALK activates JNK which , in turn, phosphorylates and activates c-Jun, resulting in uncontrolled cell cycle progression in ALCL. 293T and Jurkat (T-acute lymphoblastic leukemia) cells were transfected with a vector expressing NPM-ALK with active kinase domain (pDest40-NPM-ALK) or a construct lacking NPM-ALK kinase activity (pDest40-K210R) or empty vector. Cells were harvested at 48 hours and analyzed for protein expression by Western blot analysis and for AP-1 activity by luciferase reporter assay. Two ALK+ ALCL cell lines Karpas 299 and SU-DHL-1, found to express high levels of serine phosphorylated and total c-Jun in immunoblots, were treated with JNK (SP600125), ERK (U0126), or ALK (WHI-P154) inhibitors or were transiently transfected with siRNAs specific for JNK1 and c-Jun. Cell proliferation was assessed by MTS assay, and cell cycle was analyzed by BrdU assay or propidium iodide staining and flow cytometry. Forced expression of NPM-ALK in 293T and Jurkat cells resulted in increased levels of JNK and c-Jun phosphorylation in immunoblots and a dramatic increase in AP-1 activity. Conversely, pharmacologic inhibition of ALK activity in Karpas 299 and SU-DHL1 resulted in a concentration-dependent decrease of JNK and c-Jun phosphorylation levels. Co-immunoprecipitation studies revealed that NPM-ALK physically binds to JNK1 and its upstream activator MKK7 in ALK+ ALCL cells. Selective inhibition of JNK, but not ERK, in Karpas 299 and SU-DHL1 decreased the level of c-Jun phosphorylation in a dose-dependent manner as shown by Western blot analysis and in vitro kinase assays. Inhibition of JNK by SP600125 or silencing of the JNK1 gene by siRNA also resulted in decreased cell proliferation associated with decreased AP-1 activity, cell cycle arrest mostly at G2 phase, and up-regulation of the cyclin-dependent inhibitor p21, a transcriptional target of c-Jun. Similarly, silencing of c-Jun by specific siRNA led to decreased S-phase fraction of cell cycle, which was associated with up-regulation of p21 and downregulation of cyclin D3. These findings reveal a novel function of NPM-ALK oncoprotein, phosphorylation and activation of JNK, which may contribute to uncontrolled cell cycle progression through activation of c-Jun. Modulation of JNK or c-Jun activity may be a target for therapy in patients with ALCL.
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Elloul, Sivan, Claire Depew, Sujata Nerle, Tiffany Chen, Samuel Kaplan, Nathan Dowden, Jordi Mata-Fink, Avak Kahvejian, and Robert Deans. "Engineered Erythrocytes As an Anti-Tumor Therapy through Induction of Apoptosis or Immune-Checkpoint Inhibition." Blood 128, no. 22 (December 2, 2016): 2166. http://dx.doi.org/10.1182/blood.v128.22.2166.2166.
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Abstract Rubius Therapeutics has built a platform for producing allogeneic Red Cell Therapeutics (RCTs), genetically modified red blood cells expanded ex vivo. RCTs maintain the expected properties of normal RBCs including immunoprivilege, long circulating half life and deformability. Using lentiviral gene delivery, RCTs are able to harbor active intracellular as well as extracellular proteins, ranging from enzymes and cell targeting moieties to agonists and antibodies. RCTs represent a potentially transformational oncology platform, enabling multiple distinct modalities including tumor starvation, enhanced apoptotic signaling, and immune checkpoint inhibition, among others. The unique properties of the RCTs, including high avidity, the ability to express different moieties on and in the same cell, immunoprivilige, and extended T1/2 suggest that this platform represents the next generation of circulating cell-based therapeutics. Herein data is presented demonstrating how RCTs can be designed to target and engage specific cell types and mediate biological effects on these target cells. First, RCTs were tested for how effectively they bind to a known cell surface marker with therapeutic relevance, and whether a subsequent biological effect could be induced. We generated RCTs expressing an anti-CD20 single chain variable fragment on their surface (RCT-antiCD20) and assessed their ability to bind CD20+ lymphoma cells in vitro. We demonstrated efficient and specific binding of RCT-antiCD20 to target cells using flow cytometry and immunofluorescent microscopy. We further assessed whether this interaction could induce apoptosis by co-culturing RCT-antiCD20 cells with a panel of CD20+ human lymphoma cell lines, representing a variety of lymphoma subtypes; DoHH2 (follicular lymphoma), Ramos (Burkitt's lymphoma), Granta-519 (Mantle Cell Lymphoma) and SU-DHL-4 (diffuse large B cell lymphoma). In all cases, RCT-antiCD20 co-culture resulted in increased apoptosis relative to RCT or soluble Rituximab monoclonal antibody alone. Direct tumor cell killing in vitro was hypothesized to be more effective than monoclonal antibody alone due to the hyper-crosslinking of CD20 on the lymphoma cell. This effect was shown both by in situ demonstration of receptor clustering and by a stimulation of apoptotic pathways. Importantly, in an in vivo biodistribution study RCT-antiCD20 demonstrated strong and specific tumor pentration. These findings therefore demonstrate a novel biology for proteins expressed on RCTs and warranted testing for impact on lymphoma tumors in vivo, with lymphoma xenograft studies currently ongoing. Separately, we generated RCTs expressing on their surface antibodies against PD-1 and PD-L1 (RCT-antiPD-1 and RCT-antiPD-L1) to assess whether these RCTs could bind their respective targets and activate a robust immune response. Binding of RCT-antiPD-1 and RCT-antiPD-L1 to recombinant PD-1 and PD-L1, respectively was determined using flow cytometry. Functional activity was tested using an in vitro Jurkat cell IL-2 secretion assay. In this assay, IL-2 secretion is inhibited by incubating Jurkat cells with NHL cells (Z138) expressing PD-L1 upon stimulation with CD3/CD28 tetramers. We demonstrated that IL-2 secretion was rescued by culturing the Jurkat and Z138 cells with RCT-antiPD-1 or RCT-antiPD-L1, and not control RCT. The ability of these engineered RCTs to elicit activation was tested in a standard antigen recall assay. A robust 4-6 fold increase was demonstrated in interferon-gamma secretion of PBMC in an antigen recall assay, when co-cultured with RCT-antiPD-1 or RCT-antiPD-L1, in comparison to control PBMCs or control RCT alone. In conclusion, by using the Rubius platform for creating Red Cell Therapeutics, we were able to demonstrate that RCTs are capable of 1) engaging in specific cell-cell interactions and 2) inducing direct killing of NHL cells and 3) immune checkpoint engagement. Further studies are underway to evaluate the ability of these and other RCTs to access and kill tumor cells in vitro and in vivo. Our data support the development of RCTs as a novel class of therapeutic, enabling multiple modalities and mechanisms applicable to oncology and other indications. Disclosures Elloul: Rubius Therapeutics: Employment. Depew:Rubius Therapeutics: Employment. Nerle:Rubius Therapeutics: Employment. Chen:Rubius Therapeutics: Employment. Kaplan:Rubius Therapeutics: Employment. Dowden:Rubius Therapeutics: Employment, Equity Ownership. Mata-Fink:Rubius Therapeutics: Employment. Kahvejian:Rubius Therapeutics: Employment, Equity Ownership. Deans:Rubius Therapeutics: Employment, Equity Ownership.
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Schulz, Angela, Debra K. Czerwinski, and Ronald Levy. "B-Cell Receptors Of Follicular Lymphoma Patients Recognize Themselves." Blood 122, no. 21 (November 15, 2013): 633. http://dx.doi.org/10.1182/blood.v122.21.633.633.
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Abstract Follicular Lymphoma (FL) is the most common indolent lymphoma and is characterized by retained surface B-cell receptor (BCR) expression despite ongoing V region somatic mutation. Furthermore, every tumor has a unique BCR. Together, this suggests that the BCR is important for the survival of the malignant B cell. Recognition of a target antigen could lead to a constant stimulation of the malignant cells and serve as a driving force. Recombinant BCRs from a series of FL patients were expressed in the form of assembled proteins and all of IgG3 isotype. Recently we reported that 25% of these BCRs have binding activity against a human epithelial cell line1. In the current study we aimed to broaden the search for putative auto-antigens. As FL cells are in close contact with various peripheral blood cells (PBMC), an epitope on the surface of these cells might serve as a putative auto-antigen. Therefore, we incubated PBMCs from healthy donors with the lymphoma derived BCRs and quantified surface binding by FACS. We gated separately on B-cells, CD4 T-cells, CD8 T-cells, monocytes and natural killer (NK) cells. Thereby, we identified 7 out of 25 tumor-derived BCRs which bound to at least one cell type. Four of these strongly bound to NK, B-cells and monocytes from multiple different donors. None of the tested antibodies bound to T-cells. We tested the four reactive BCRs against B-cell lines (Daudi, Raji, Ramos, DHL-4, FL-18 and Reh) and one T-cell line (MOLT-4). We found that Daudi and to a lesser extent Raji but none of the other cell lines were positive for the same lymphoma derived BCRs as PBMCs. Because of the observed binding pattern we hypothesized that a common protein modification might be the target. We therefore treated the cell lines with tunicamycin, an inhibitor for N-linked glycosylation, in order to test if sugars might be the targets of the lymphoma derived BCRs. Surprisingly, this led to stronger binding of the same BCRs. it is known that de-glycosylation of Fc receptors increases Fc binding, suggesting that Fc receptors might be the target. In line with this hypothesis Daudi cells have Fc receptors but Ramos cells do not. Moreover, T-cells are the only PBMCs which do not express Fc receptors. Therefore, we tested Fc receptors directly as targets. Recombinant Fcγ1 (CD64) and Fcγ3 (CD16) were positive in ELISA tests with the four recombinant lymphoma-derived BCRs. In addition, blocking Fcγ2 (CD32) and Fcγ3 receptors on PBMCs before staining with lymphoma derived BCRs resulted in a signal reduction. In order to define if the Fc part of these BCRs is passively bound by Fc receptors or if the Fc receptor is recognized as specific target by the variable BCR regions we produced F(ab)2 fragments. Staining PBMCs with these abolished binding, suggesting that the Fc region of the lymphoma derived BCRs is bound necessary for cellular binding. As all BCRs have the same IgG3 constant region they should all bind to Fc receptors through their Fc regions with the same affinity. In vivo Fc receptor bearing cells are not activated by the binding of a single antibody but only when an antibody cluster is formed e.g. the antibody-coated surface of a pathogen. We therefore hypothesized that the BCRs which bound Fc receptors exist as clusters. This would be conceivable if the BCRs recognized a part of them-selves as target which subsequently would lead to dimer or oligomerization. Indeed, a Fc binding BCR which was coated on an ELISA plate could be detected with a biotinylated counterpart of itself. This same phenomenon was recently described to be the case for chronic lymphocytic leukemia2. Experiments are ongoing to elucidate which region of the BCRs are recognized as targets and for which percentage of them for which this is true. In conclusion these findings suggest a new target for the BCR of FL cells whose constant presence in its microenvironment might represent a novel mechanism of chronic BCR stimulation. 1Sachen KL et al., Blood 2012, 2Dühren von Minden M et al., Nature 2012 Disclosures: No relevant conflicts of interest to declare.
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Bianchi, Giada, Peter G. Czarnecki, Aldo M. Roccaro, Antonio Sacco, Yawara Kawano, Mehmet K. Samur, Annamaria Gulla, et al. "Roundabout 1 (ROBO1)/SLIT2 Is a Novel Signaling Pathway in Multiple Myeloma Promoting Survival and Bone Marrow Niche Interaction." Blood 128, no. 22 (December 2, 2016): 485. http://dx.doi.org/10.1182/blood.v128.22.485.485.
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Abstract Introduction Molecular mechanisms of multiple myeloma (MM) pathogenesis are unknown, but a role for bone marrow (BM) niche in supporting MM is well established. The transmembrane receptor Roundabout 1 (ROBO1) has been historically annotated as a tumor suppressor gene in solid cancer, and more recently myelodysplastic syndrome. High ROBO1 expression was identified as a poor prognostic factor in newly diagnosed MM; however, its function in MM is unknown. Material and Methods We analyzed ROBO1 and SLIT2 expression in human MM cell lines (HMCL) and a panel of human cancer cell lines via western blot (WB). ROBO1/SLIT2 expression in primary BM samples was assessed via immunohistochemistry and/or WB. We used short hairpin RNA (shRNA) for stable ROBO1 knock down (KD). We designed and cloned single guide RNA (sgRNA) into pSpCas9(BB)-2A-GFP vector and developed CRISPR-Cas9-mediated ROBO1 knock out (KO) 293T and OPM2 clones. WT and KO ROBO1 OPM2 cells were used for standard viability, proliferation, and adhesion assays. Full length (FL) and mutant ROBO1 isoforms devoid of extracellular or intracellular domain, with a C-terminus triple FLAG tag were designed and cloned into pCDH lentiviral vectors. WT and ROBO1 KO OPM2 were injected in the BM cavity of femoral bones harvested from donor SCID mice, and implanted subcutaneously into recipient SCID mice to study ROBO1 KO in vivo. Results HMCL, primary MM and BM stromal cells (MM-BMSC) from patients, but not healthy donor BM-resident plasma cells and BMSC, express ROBO1 and SLIT2. Across a panel of myeloid and lymphoid cancer cell lines, high ROBO1 expression was detected only in SU-DHL-4 and BCWM1 cell lines. In newly diagnosed MM patients enlisted in the IMF170 database, high ROBO1 portended worse overall survival compared to low ROBO1 expression (P=0.018). Sh-RNA mediated ROBO1 KD was profoundly and specifically cytotoxic to HMCL, with no cytotoxicity observed in high-ROBO1 expressing SH-SY5Y and HeLa cancer cell lines. Consistent with on target effect, ROBO1 shRNA was not cytotoxic for non-ROBO1 expressing cancer cell lines. To study the functional consequences of ROBO1 loss in MM, we performed CRISPR-Cas9-mediated ROBO1 KO in HMCL. We were unable to establish biallelic ROBO1 KO H929 or U266 clones, suggesting that biallelic loss may be lethal in these HMCL, but created two clones of ROBO1 KO OPM2 cells. In vitro, ROBO1 KO and WT OPM2 cells have comparable viability and proliferation rate, even in the presence of MM-BMSC. However, ROBO1 KO OPM2 cells have profound defects in adhesion to MM-BMSC and BM endothelial cell lines (BMEC). Lentiviral transduction of OPM2 ROBO1 KO cells with FL-ROBO1 addback lentiviral construct completely rescues the BMEC adhesion defect, confirming that loss of ROBO1 is responsible for the defective adhesive phenotype. Interestingly, treatment with heparin abolishes the rescue effect of FL-ROBO1, implying that surface heparan sulfate proteoglycans (HSPG) are necessary for ROBO1-mediated MM-BMEC adhesion. Consistent with a role for ROBO1 in MM-BM niche interaction, ROBO1 KO OPM2 cells show higher failure to engraft in the bone implants compared to ROBO1 WT OPM2 (bone implants without MM engraftment: 5/15 versus 1/15, respectively, P=0.067). PET-CT scan of study mice shows ROBO1 KO tumors to be significantly smaller compared to ROBO1 WT (mean volume 1.2 cm3 vs. 2.9 cm3, P=0.003), suggesting a profound in vivo proliferation/survival defect of ROBO1 KO OPM2. Summary and Conclusions Our data demonstrate that ROBO1 and SLIT2 are highly expressed in HMCL and in the BM of MM patients, but not healthy controls. ROBO1 expression correlates with worse OS in newly diagnosed MM patients; and ROBO1 KD is selectively cytotoxic for HMCL, but not high-ROBO1 expressing SH-SY5Y and HeLa cells. CRISPR-Cas9-mediated ROBO1 KO significantly impairs OPM2 cell adhesion to BMEC and MM-BMSC, which is completely reversed by adding back FL-ROBO1. In vivo, ROBO1 KO OPM2 cells show impaired engraftment to bone implants and significantly decreased growth rate compared to ROBO1 WT OPM2. Altogether, our data demonstrate that ROBO1 is a novel pro-survival protein in MM, mediating MM-BM niche interaction and suggest the therapeutic potential of targeting ROBO1/SLIT2 pathway in MM. Proteomic and RNA-sequencing studies on the downstream signaling effectors of ROBO1/SLIT2 are currently ongoing to elucidate the molecular mechanisms of pathway activation in MM. Disclosures Roccaro: Takeda Pharmaceutical Company Limited: Honoraria. Ghobrial:Takeda: Honoraria; Novartis: Honoraria; Celgene: Honoraria, Research Funding; Noxxon: Honoraria; Amgen: Honoraria; BMS: Honoraria, Research Funding. Anderson:Oncoprep: Equity Ownership; Acetylon: Equity Ownership; Celgene: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees; Gilead: Membership on an entity's Board of Directors or advisory committees; Gilead: Membership on an entity's Board of Directors or advisory committees; Oncoprep: Equity Ownership; Acetylon: Equity Ownership; Millennuim: Membership on an entity's Board of Directors or advisory committees; Millennuim: Membership on an entity's Board of Directors or advisory committees; C4 Therapeutics: Equity Ownership; C4 Therapeutics: Equity Ownership; Bristol Myers Squibb: Membership on an entity's Board of Directors or advisory committees; Bristol Myers Squibb: Membership on an entity's Board of Directors or advisory committees.
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"Training returns DHL Express to the road of exceptional customer service." Human Resource Management International Digest 21, no. 5 (July 12, 2013): 26–29. http://dx.doi.org/10.1108/hrmid-05-2013-0041.
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"Phillip Madanire." Mosaic, no. 32 (November 29, 2004). http://dx.doi.org/10.7238/m.n32.0428.
Full textAbstract:
Phillip Madanire es Director Creativo Asociado en OgilvyOne Worldwide, New Cork, EE.UU. Responsable de los servicios de marketing interactivo y desarrollo de Internet para clientes como IBM, American Express, Ameritrade, Sprite, DuPont, Deloite, DHL, Kodak, AT&T, ONDCP, Motorola, etc. Phillip reside en la ciudad de Nueva York. Nació en Zimbabwe y ha estudiado en las mejores escuelas de India y Estados Unidos.
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Fernández Cabaleiro, Begoña. "Chillida y la realidad : ¿un nuevo romanticismo?" Espacio Tiempo y Forma. Serie VII, Historia del Arte, no. 11 (January 1, 1998). http://dx.doi.org/10.5944/etfvii.11.1998.2323.
Full textAbstract:
El arte del romanticismo es la expresión de una sensibilidad que se libera de las reglas establecidas con el fin de buscar nuevas fuentes de inspiración. Esto lleva al artista a buscar nuevos caminos a través de la naturaleza. A finales del siglo XX el hombre vive una situación de vértigo. El arte surge como una vía de transfiguración. Eduardo Chillida parte de la realidad. La relee y la expresa de un modo nuevo. Agua, luz, espacio, sonido; hombre y libertad; espíritu y materia son de nuevo descubiertas y manifestadas en su obra.Romanticize art was a way of sensibility without regulation; It wanted to look for new inspiration ways. The artist looked into the Nature. At the end of XXth century, the man lives a difficult situation.The art borns as a way of change. Eduardo Chillida begins in the reality. He studies it and he makes it in a new way. Water, light, space, sound; Man and liberty; mind and matter are discovered again in his work.