Immunostimulation by OX40 Ligand Transgenic Ewing Sarcoma Cells

Interleukin-2 (IL-2) transgenic Ewing sarcoma cells can induce tumor specific T and NK cell responses and reduce tumor growth in vivo and in vitro. Nevertheless, the efficiency of this stimulation is not high enough to inhibit tumor growth completely. In addition to recognition of the cognate antigen, optimal T cell stimulation requires signals from so-called co-stimulatory molecules. Several members of the tumor necrosis factor superfamily (TNFSF) have been identified as co-stimulatory molecules that can augment anti-tumor immune responses. OX40 (CD134) and OX40 ligand (OX40L = CD252; also known as tumor necrosis factor ligand family member 4) is one example for such receptor/ligand pair with co-stimulatory function. In the present investigation we generated OX40L transgenic Ewing sarcoma cells and tested their immuno-stimulatory activity in vitro. OX40L transgenic Ewing sarcoma cells showed preserved expression of Ewing sarcoma associated (anti)gens including lipase member I (LIPI), cyclin D1 (CCND1), cytochrome P450 family member 26B1 (CYP26B1) and the Ewing sarcoma breakpoint region 1-friend leukemia virus integration 1 (EWSR1-FLI1) oncogene. OX40L expressing tumor cells showed a trend for enhanced immune stimulation against Ewing sarcoma cells in combination with IL-2 and stimulation of CD137. Our data suggest that inclusion of the OX40/OX40L pathway of co-stimulation might improve immunotherapy strategies for treatment of Ewing sarcoma

Oxidative damage control in a human (mini-) organ: Nrf2 activation protects against oxidative stress-induced hair growth inhibition

The in situ control of redox insult in human organs is of major clinical relevance, yet remains incompletely understood. Activation of Nrf2, the “master regulator” of genes controlling cellular redox homeostasis, is advocated as a therapeutic strategy for diseases with severely impaired redox balance. It remains to be shown whether this strategy is effective in human organs, rather than isolated human cell types. We have therefore explored the role of Nrf2 in a uniquely accessible human (mini-) organ, human scalp hair follicles (HFs). Microarray and qPCR analysis of human HFs following Nrf2 activation using sulforaphane identified the modulation of phase II metabolism, ROS clearance, the pentose phosphate pathway and glutathione homeostasis. Nrf2 knockdown (siRNA) in cultured human HFs confirmed the regulation of key Nrf2 target genes (i.e. HO-1, NQO1, GSR, GCLC, ABCC1, PRDX1). Importantly, Nrf2 activation significantly reduced ROS levels and associated lipid peroxidation. Nrf2 pre-activation reduced oxidative stress-stimulated (H2O2 or menadione) premature catagen and hair growth inhibition, significantly ameliorated the H2O2-dependent increase in matrix keratinocyte apoptosis and reversed the ROS-induced reduction in proliferation. This study thus provides direct evidence for the crucial role of Nrf2 in protecting human organ function (i.e. scalp HFs) against redox insult

The first European interdisciplinary Ewing sarcoma research summit

The European Network for Cancer Research in Children and Adolescents (ENCCA) provides an interaction platform for stakeholders in research and care of children with cancer. Among ENCCA objectives is the establishment of biology-based prioritization mechanisms for the selection of innovative targets, drugs, and prognostic markers for validation in clinical trials. Specifically for sarcomas, there is a burning need for novel treatment options, since current chemotherapeutic treatment protocols have met their limits. This is most obvious for metastatic Ewing sarcoma (ES), where long term survival rates are still below 20%. Despite significant progress in our understanding of ES biology, clinical translation of promising laboratory results has not yet taken place due to fragmentation of research and lack of an institutionalized discussion forum. To fill this gap, ENCCA assembled 30 European expert scientists and five North American opinion leaders in December 2011 to exchange thoughts and discuss the state of the art in ES research and latest results from the bench, and to propose biological studies and novel promising therapeutics for the upcoming European EWING2008 and EWING2012 clinical trials

Expression and regulation of the endogenous retrovirus 3 in Hodgkin's lymphoma cells

Human endogenous retroviruses (ERV) are an integral part of our genome. Expression of ERV is usually switched off but reactivation of ERV has been observed in varying human diseases including cancer. Recently, reactivation of ERV associated promoters in Hodgkin’s lymphoma (HL) cells has been described. Despite relatively good prognosis, not all patients with HL can be cured with the established therapy and this therapy is associated with severe late side effects. Therefore, new targets are required for the development of future treatment strategies. Reactivated ERV might represent such target structures. Therefore, we asked which ERV loci are expressed in HL cells. Using DNA microarray analysis, we found no evidence for a general activation of ERV transcription in HL cells. In contrast, we observed down-regulation of ERV3, an ERV with potential tumor suppressor function, in HL cells in comparison to normal blood cells. Interestingly, ERV3 was also differentially expressed in published DNA microarray data from resting versus cycling B cells. Treatment of HL cells with the histone deacetylase inhibitor vorinostat strongly up-regulated ERV3 expression. In addition, we observed up-regulation in HL cells after treatment with hypoxia-mimetic cobalt(II) chloride. Like vorinostat, cobalt(II) chloride inhibited cell growth of HL cells. Our results suggest that cell cycle inhibition of HL cells is accompanied by up-regulation of ERV3

Bilder in Texten – Analyse von Eingewöhnungskonzepten mit der dokumentarischen Methode

Der Beitrag fokussiert das methodische Vorgehen bei der Analyse von Eingewöhnungskonzepten als Konzeptpapieren in Schrift und Bild, wie sie von Kindertageseinrichtungen selbst erstellt bzw. an Eltern und weitere Adressat_innen ausgegeben werden. Rahmen der Analyse bildet das Forschungsprojekt „Eingewöhnungskonzepte zwischen Programmatik und Praxis“, in dem untersucht wird, wie pädagogische Fachkräfte in und mit solchen Konzepten ihren gesellschaftlichen Auftrag zur Betreuung der Kinder interpretieren und wie sie öffentliche und familiale Kleinkindbetreuung dabei ins Verhältnis setzen. Die Beispielanalyse konzentriert sich hier vornehmlich auf das Bildmaterial in solchen Dokumenten, die von Einrichtungen im Internet publiziert wurden. Mit der Dokumentarischen Methode – die als etablierte Methode bislang vor allem zur Analyse von Interviews und Gruppendiskussionen, aber auch Bilddokumenten eingesetzt wird – wird dafür ein Analyseverfahren der rekonstruktiven Sozialforschung genutzt, um empirischen Zugang zur konzeptionellen Ebene frühpädagogischer Praxis zu gewinnen. Durch die schrittweise nachvollziehbare formulierende und reflektierende Interpretation einer Abbildung auf dem Deckblatt eines Eingewöhnungskonzepts sowie die abschließende komparative Analyse wird die spezifische Art der Herstellung und Kontextualisierung der Bilder herausgearbeitet. Deutlich werden die Wissensbestände, die in den Konzepten eine bedeutsame Rolle spielen und wie sie bei der Thematisierung der Eingewöhnung – z. B. gegenüber den Eltern – zur Geltung gebracht werden. DOI 10.18442/09

Targeting the epigenetic readers in Ewing Sarcoma inhibits the oncogenic transcription factor EWS/Fli1.

Ewing Sarcoma is a rare bone and soft tissue malignancy affecting children and young adults. Chromosomal translocations in this cancer produce fusion oncogenes as characteristic molecular signatures of the disease. The most common case is the translocation t (11; 22) (q24;q12) which yields the EWS-Fli1 chimeric transcription factor. Finding a way to directly target EWS-Fli1 remains a central therapeutic approach to eradicate this aggressive cancer. Here we demonstrate that treating Ewing Sarcoma cells with JQ1(+), a BET bromodomain inhibitor, represses directly EWS-Fli1 transcription as well as its transcriptional program. Moreover, the Chromatin Immuno Precipitation experiments demonstrate for the first time that these results are a consequence of the depletion of BRD4, one of the BET bromodomains protein from the EWS-Fli1 promoter. In vitro, JQ1(+) treatment reduces the cell viability, impairs the cell clonogenic and the migratory abilities, and induces a G1-phase blockage as well as a time- and a dose-dependent apoptosis. Furthermore, in our in vivo model, we observed a tumor burden delay, an inhibition of the global vascularization and an increase of the mice overall survival. Taken together, our data indicate that inhibiting the BET bromodomains interferes with EWS-FLi1 transcription and could be a promising strategy in the Ewing tumors context

The first European interdisciplinary Ewing sarcoma research summit

The European Network for Cancer Research in Children and Adolescents (ENCCA) provides an interaction platform for stakeholders in research and care of children with cancer. Among ENCCA objectives is the establishment of biology-based prioritization mechanisms for the selection of innovative targets, drugs, and prognostic markers for validation in clinical trials. Specifically for sarcomas, there is a burning need for novel treatment options, since current chemotherapeutic treatment protocols have met their limits. This is most obvious for metastatic Ewing sarcoma (ES), where long term survival rates are still below 20%. Despite significant progress in our understanding of ES biology, clinical translation of promising laboratory results has not yet taken place due to fragmentation of research and lack of an institutionalized discussion forum. To fill this gap, ENCCA assembled 30 European expert scientists and five North American opinion leaders in December 2011 to exchange thoughts and discuss the state of the art in ES research and latest results from the bench, and to propose biological studies and novel promising therapeutics for the upcoming European EWING2008 and EWING2012 clinical trials

Immunogenicity of the Envelope Surface Unit of Human Endogenous Retrovirus K18 in Mice

The triggers for the development of multiple sclerosis (MS) have not been fully understood to date. One hypothesis proposes a viral etiology. Interestingly, viral proteins from human endogenous retroviruses (HERVs) may play a role in the pathogenesis of MS. Allelic variants of the HERV-K18 env gene represent a genetic risk factor for MS, and the envelope protein is considered to be an Epstein–Barr virus-trans-activated superantigen. To further specify a possible role for HERV-K18 in MS, the present study examined the immunogenicity of the purified surface unit (SU). HERV-K18(SU) induced envelope-specific plasma IgG in immunized mice and triggered proliferation of T cells isolated from these mice. It did not trigger phenotypic changes in a mouse model of experimental autoimmune encephalomyelitis. Further studies are needed to investigate the underlying mechanisms of HERV-K18 interaction with immune system regulators in more detail