[FG] Schär PrimoHead of Research Unit Prof. Dr.Primo Leo SchärOverviewMembersPublicationsProjects & CollaborationsProjects & Collaborations OverviewMembersPublicationsProjects & Collaborations Projects & Collaborations 19 foundShow per page10 10 20 50 Validation of therapeutic approaches targeting epigenetic enzymes in a pre-clinical model of congenital myopathies. Research Project | 2 Project MembersSkeletal muscle contraction starts when Ca2+ is released from the sarcoplasmic reticulum via the ryanodine receptor (RyR1). Defects in Ca2+ signaling, often due to RYR1 mutations, cause neuromuscular disorders like malignant hyperthermia and congenital myopathies. To study recessive RYR1 mutations, a mouse model (dHT) was created with specific mutations found in a child with multi-minicore disease (MmD). Treatment of dHT mice with drugs targeting epigenetic enzymes (HDACs and DNMT inhibitors) improved muscle strength and function. The study aims to explore the biochemical changes these drugs trigger, potentially leading to clinical trials for treating congenital myopathies. Air Pollution and Effects on Lung Functional Development and Respiratory Morbidity in At-Risk Infants Research Project | 4 Project MembersBACKGROUND AND RATIONALE: This is a direct continuation of SNF 182871/1, which investigated the impact of early-childhood environmental factors on lung functional growth and consequences for later respiratory morbidity in healthy term infants. We previously demonstrated that even low-level air pollution exposure during pregnancy and early childhood is associated with impaired lung functional growth in infancy and early childhood. Although the mechanisms are still unclear, they could be related to lung functional growth deficits or remodeling of the lung due to changes in the intrauterine environment. Air pollution is known to induce oxidative stress response and related autophagy and cellular senescence mechanisms, potentially playing a role in pollution-related lung pathology and in remodeling. As novel preliminary evidence in SNF 182871/1, we recently found that, in the cord blood of human infants, autophagy-related biomarkers are correlated with remodeling biomarkers. We also found that air pollution exposure during pregnancy is associated with biomarkers of autophagy and remodeling in the cord blood of healthy term infants. Interestingly, these mechanisms also play an important role in fetal development and preterm birth, and may thus theoretically contribute to the susceptibility of infants-and particularly preterm infants-to oxidative stress and air pollution effects. Indeed, as first evidence from SNF 182871/1, we also found an enhanced impact of air pollutants on lung function impairment of preterm infants. Furthermore, our own preliminary human data show that markers of autophagy, and remodeling already have significant differences between the cord blood of preterm infants compared to term infants at birth prior to early postnatal injury. Bringing this together, we hypothesize that the interaction of oxidative stress response, autophagy and remodeling could be a key mechanism involved in the complex host-environment interaction determining lung functional growth and related respiratory morbidity. Moreover, this response could be different in infants at risk for chronic respiratory symptoms, such preterm infants, infants born from asthmatic mothers or infants exposed to high levels of air pollution during pregnancy. OVERALL OBJECTIVES: We aim to expand the ongoing BILD cohort of (i) term infants with two risk subgroups, (ii) infants born preterm, and (iii) infants born to asthmatic mothers, and we will investigate the differences in response to prenatal air pollution in relation to the above key mechanisms. SPECIFIC AIMS: In comparison to healthy term infants, we will investigate in study phase 1, (i) whether the increased susceptibility of infants to prenatal air pollution in these three risk groups is related to differences in markers of oxidative stress response, autophagy, and remodeling in cord blood and in study phase 2, (ii) whether these pollution-related cord blood profiles are correlated to lung functional development and subsequent symptoms in the first year of life (primary outcomes) and at school age (secondary outcomes). We will replicate these findings in other birth cohorts from collaborators (Germany, Australia) with comparable outcome measures. METHODS: In our prospective BILD birth cohort of 1000 unselected healthy term infants, 400 preterm infants, and 200 infants from asthmatic mothers we will (i) estimate indoor and outdoor air pollution exposure during pregnancy and in early infancy, (ii) assess family, obstetric and birth history, cord-molecular biomarkers (metabolomics, gene expression, proteins), and infant lung function shortly after birth (including exhalomics) and at 6 years of age, as well as respiratory symptoms in the first year of life and at school age. EXPECTED RESULTS AND IMPACT: We expect a 26.03.2021 18:35:26 Page - 14 - significant correlation between air pollution exposure and oxidative stress response and lung remodeling in newborns with effects on lung function and clinical outcomes, the latter effects enhanced in the risk groups. Particularly for these risk groups, today's air pollution may already result in lung remodeling and subsequent impaired lung functional growth even at this early stage of life. Since early-life lung functional impairment often persists until school age and even late adulthood, it is a previously described early-life risk factor known to be associated with asthma in children and chronic obstructive respiratory airway diseases in the elderly. Thus, early-life environmental injury has a potentially very relevant impact on future global respiratory health, with unpredictable costs. We are one of the first groups to look into the impact of these air-pollution-induced mechanisms on oxidative stress response and lung remodeling, subsequent impairment of lung functional growth, and resulting human lung disease. Better understanding of these mechanisms might help the development of preventative and therapeutic strategies, particularly for at-risk infants. Impact of mobile communication signals on the regulation of neural differentiation Research Project | 4 Project MembersOur project aims at studying possible effects of radiofrequency electromagnetic fields (RF-EMF) on neuronal differentiation and related cellular pathways known to be involved in neurodegeneration and associated diseases. Focusing on pathways playing a role in neuronal differentiation and degeneration, we will investigate RF-EMF effects on distinct neuronal cell populations before and during differentiation and identify molecular pathways involved in a hypothesis-free genomic approach as well as in a hypothesis-driven approach. These goals will be achieved by applying cell culture-based experimentations including neuron-like and neuronal stem/progenitor cells. Progression of differentiation and phenotypic characterization will be assessed by fluorescence microscopy of stem cell and neuronal markers combined with high-content analysis and evaluation of morphology (i.e. neurite outgrowth). These analyses will be complemented by the assessment of key players of cellular pathways (e.g. the ERK/MAP-K, PI3-K/Akt, Wnt/β-catenin) underlying the morphological changes such as neurite outgrowth in a quantitative manner (e.g. by Western blotting and FRET-biosensor). Mitochondrial dysfunction during neurodegeneration was shown to increase the production of reactive oxygen species (ROS). Oxidative stress is a frequent early condition in the pathogenesis of neurodegenerative diseases, is often observed in cells upon EMF exposure and has an evident potential to compromise genetic as well as epigenetic stability. Mitochondrial integrity, an important indicator of neuronal aging and degeneration will be investigated quantitatively. To assess the impact of RF-EMF exposure on the composition and behavior of the differentiating neural cell population, gene expression profiling (transcriptome) of single cells before and after differentiation will be performed. Cluster analysis of gene expression profiles will describe the dynamics of the cell population and potentially result in the identification of target regions with altered epigenetic modifications. Target-specific quantitation of DNA methylation and histone modifications at regulatory elements (promoter, enhancer) of identified genes, markers of neural stem cell and the neuronal lineage will be analyzed by pyrosequencing of bisulfite-converted DNA and by chromatin immunoprecipitation (ChIP). This project will provide a significant and critical insight into the adverse effects of exposure to modulated RF-EMF as used for mobile communication (GSM) on signaling cascades and physiology as well as on morphological and epigenetic characteristics of neural cells in vitro . BERENIS Berater Research Project | 1 Project MembersDas BAFU hat als Umweltfachstelle des Bundes die Aufgabe, die Forschung über gesundheitliche Auswirkungen nichtionisierender Strahlung von Infrastrukturanlagen wie auch diesbezügliche Alltagserfahrungen zu verfolgen und die Ergebnisse zu bewerten, sowie die Öffentlichkeit über den Stand der Wissenschaft und der Erfahrung zu informieren. Sofern neue Ergebnisse dies erfordern, stellt es dem Bundesrat Antrag auf eine Anpassung der Immissionsgrenzwerte der NISV. Andernfalls muss es - insbesondere gegenüber technologiekritischen Kreisen - jederzeit begründen können, weshalb die geltenden Immissionsgrenzwerte nicht angepasst werden müssen. Die Bewertung muss Aussagen darüber machen, wie sicher biologische Effekte nachgewiesen sind, ob sie für die Gesundheit relevant sind und wie viele Menschen gegebenenfalls betroffen sind. Zur Bewertung gehört auch eine Früherkennung potenzieller Risiken. Dem Bundesrat wird in gerichtlichen Beschwerdeverfahren regelmässig vorgeworfen, die Grenzwerte der NISV nicht nach dem Stand der Wissenschaft oder der Erfahrung festgesetzt zu haben. Beschwerdeführer legen eigene Bewertungen des Standes der Wissenschaft vor, um wesentlich strengere Grenzwerte zu fordern. Das Bundesgericht hat diese Begehren bisher mit dem Hinweis zurückgewiesen, dass es Aufgabe der Fachbehörden des Bundes sei, den Stand der Wissenschaft zu evaluieren, und dass das BAFU dieser Aufgabe nachkomme. Die zuständige Sektion NIS des BAFU hat bisher das Schweiz. Tropen- und Public Health Institut Basel (Swiss TPH) mit der Bewertungsarbeit beauftragt, und kurze Zeit auch das Institut für Biomedizin der Universität Basel. Das TPH hat die wissenschaftlichen Publikationen zu Humanstudien möglichst vollständig erfasst, bewertet und in der Datenbank ELMAR öffentlich zugänglich gemacht. Daraus hat es mehrere thematische Syntheseberichte erstellt, welche das BAFU in seinen Publikationsreihen publiziert hat. Diese Bewertungsarbeit konzentrierte sich auf Humanstudien. Der Fokus soll nun auf experimentelle (Tiere und Zellen) sowie mechanistische Studien ausgeweitet und die Verantwortung für die Bewertung breiter abgestützt werden, um eine umfassendere Sicht des wissenschaftlichen Kenntnisstandes zu erhalten. Zu diesem Zweck wird eine beratende Expertengruppe eingesetzt. Analysis of TDG Dependent Ribonucloprotein Complex formation on Long Noncoding RNA by SILAC Research Project | 1 Project MembersThe generation of a diversity of cells with different functions is crucial for multicellular organisms. The individual cell identity relies on a selective readout of the genome achieved with the help of epigenetic mechanisms which direct the transcriptional programs to cell-type specific endpoints. Working at the level of chromatin, these mechanisms orchestrate gene transcription by deposition, exchange or removal of epigenetic modifications at histones as wells as in the DNA. Inaccuracy of the epigenetic system not only impedes successful establishment of cell identity but may also have deleterious effects on tissue homeostasis. This is best exemplified by epigenetically unstable cancers which frequently show aberrant patterns of CpG methylation. Thus, a faithful control of the epigenetic machinery is critical for the preservation of correct cell identity. We have identified Thymine DNA glycosylase as a potential epigenetic proof-reader in CpG rich promoters of genes in active and bivalent chromatin. TDG helps to maintain these regions in an unmethylated state by initiating a DNA repair process at sites of erroneous cytosine methylation. Research of the last years has shed light on how TDG dependent active demethylation may occur at the molecular level; yet, the ways how this process is regulated and targeted to its site of actions remain obscure. Hints on how this may be achieved come from knowledge obtained for long noncoding RNAs. Serving as scaffolds, some of these RNAs have been shown to play a central role for the regulation of epigenetic processes. Together with our own preliminary data showing association of TDG with a variety of lncRNAs this suggests that these RNAs may be important for TDG mediated active DNA demethylation. Intriguingly, previously identified interaction partners of TDG, such as e.g. nuclear receptors have been reported to associate with some of the identified TDG bound lncRNAs, especially steroid receptor RNA activator 1. The work proposed herein will follow-up on this important question and investigate the TDG dependent composition of ribonucleoprotein complex by SILAC using SRA1 as a model lncRNA. Sub-cellular targeting microscopy - Signaling in Development and Oncology Research Project | 6 Project MembersIn the past decade, research in development and oncology has uncovered an impressive number of relevant signaling pathways. While a qualitative understanding of signals, and their connection to cellular outputs has been established, plasticity and feedback of signaling networks remain obscured. A better understanding of dynamic and locally constrained signaling events driving organ development and disease progression requires access to refined subcellular probe detection. The availability of optogenetic and chemical biology tools provides novel opportunities, but requires dedicated microscopy equiment. For this reason, six projects at the Department of Biomedicine (DBM) and the Biocenter (BC) of the Univerisity of Basel i) localized lipid signaling in disease (M. Wymann, DBM); ii) dynamic subcellular Wnt/b-catenin signaling in epithelial mesenchymal transition (G. Christofori, DBM); iii) DNA dynamics and confined epigenetic plasticity (P. Schär, DBM); iv) real-time monitoring of Sonic Hedgehog and Bone Morphogenetic Protein gradients in limb buds (R. Zeller, DBM); v) ultrastructural analysis of neuronal stem cell control (V. Taylor, DBM); and vi) molecular mechanisms determining the development of vascular networks (M. Affolter, BC), illustrate the need of the requested "subcellular targeting microscopy" equipment. The core of the platform is a highly sensitive microlens-enhanced spinning disk microscope linked to FRAP, ablation, and multiple excitation laser lines, and an integrated TIRF module to monitor plasma membrane events. Through its integration into the BioOptics core facility at the DBM, the subcellular targeting platform will be accessible to >500 regional researchers at the University of Basel, DBM, FMI, D-BSSE, Fachhochschule, etc. An image storage and analysis pipeline with remote user access capabilitiy is in place, to allow seamless operation and output. The requested equipment will allow the use of genetically encoded opto-genetic proteins, proteins tagged for reactivity with chemical inducers of dimerisation (CIDs), and the possibility to perform FRAP/TIRF and FRAP/confocal microscopy, and will greatly enhance the possibilities to manipulate and track subcellular localization of target proteins. The insights gained by these experimental approaches will be critical for a better understanding of dynamic biological processes, and will spur the design of innovative therapeutic approaches to counteract resistance mechanisms in oncology. Characterization of the Ewing's sarcoma protein's involvement in the maintenance of genomic stability Research Project | 3 Project MembersConsidering that a variety of genetic abnormalities typically occur in a given tumor, it is striking that some cancers exhibit highly characteristic genomic changes. They consist in chromosome fusions that occur at very specific chromosomal locations, one of which is always the locus of a FET protein. FET stands for the family of fused in sarcoma (FUS), Ewing sarcoma (EWS) and TATA box binding protein associated factor 15 (TAF15) proteins. The most common fusion proteins are EWS-FLI1 in Ewing sarcoma, FUS-CREB3L1 in low-grade fibromyxoid sarcoma and TAF15-NR4A3 in extraskeletal myxoid chondrosarcoma. Although normal FET proteins bind RNA, this activity is absent in the fusion proteins. Whether this loss is relevant for the development of the tumor is not known. In a previous study we mapped the RNA targets of EWS. To our surprise, we found that EWS binds RNAs that come from genomic loci that are prone to 'genomic instability'. We further obtained evidence that EWS helps the transcription of DNA into messenger RNAs, facilitating the progression of the RNA polymerization enzyme (RNAPII) through loci that are prone to form stable structures that impede transcription. In this project we would like to characterize the relationship between EWS and RNAPII-dependent transcription, and the consequences at the cellular level of the long-term reduction in EWS expression. We will employ cells and cell lines in which we can reduce EWS expression in a controlled manner. We will determine the rate at which the RNA polymerase II elongates transcripts genome-wide and the frequency with which lesions in the DNA that are known as DNA double-strand breaks occur when EWS is limiting. Furthermore, we will investigate the long-term consequences of reduced EWS expression on cell morphology, viability and functionality. Our study could thus contribute to an improved understanding of the pathogenic mechanisms of Ewing's sarcoma. Cellular and molecular effects of pulsed electromagnetic fields Research Project | 3 Project MembersInterfering with cell proliferation (growth control) is an established and successful concept of both, conventional and targeted cancer therapy, but often the therapeutic choice is a double edged sword. Treatments interfering with cell proliferation are potentially harmful because they are severely toxic and often cause undesirable short and long side-effects on different tissues, adversely affecting the patient's well-being. In this respect, the observation that low intensity pulsed electromagnetic fields (PEMFs) interfere with the proliferation of cancer cells may reflect a promising strategy for future cancer therapeutics. Importantly, non-cancerous tissue cells do not seem to be affected and stem cells respond by increased proliferation and and/or differentiation. Molecular targets and mechanisms underlying this potential therapeutic effect of PEMFs are poorly understood to date, but there are indications pointing to an involvement of signalling cascades, mitotic failure and induced apoptosis. The investigation and characterization of these mechanisms at the cellular and molecular level is the focus of the proposed project. It involves cell culture-based experimental approaches using advanced microscopy and molecular biological means to explore the PEMF-induced network of anti-proliferative signals and response of cancer cells from different origin in comparison to differentiated and undifferentiated cells. Key readouts will be parameters of cell proliferation (i.e. cell viability, proliferation and apoptosis), mitotic and cell phase progression and growth-, stress- and checkpoint signalling activities. The proposed experiments aim to reveal whether PEMF-mediated reduction of cell proliferation is a common phenomenon of cancer cells or rather restricted to a spectrum of responsive cancers. Designed to identify the responsive targets, pathways and structures, they will extend the current understanding about how cells react instantly to PEMF exposure and how these responses are then transduced to produce the cellular consequences observed. The proposed work will thus provide insight into how PEMFs and possibly other types of electromagnetic signals interact with cellular processes to disturb cell proliferation and, thereby, a mechanistic framework for a rational design of therapeutic applications. DNA Repair, Epigenetic Stability, and CpG Island Hypermethylation in Colorectal Tumorigenesis Research Project | 1 Project MembersNo Description available Impact of DNA ligase IV on genome stability during DNA replication Research Project | 1 Project MembersResearch grant to Dr. Olivier Fritsch 12 12 OverviewMembersPublicationsProjects & Collaborations
Projects & Collaborations 19 foundShow per page10 10 20 50 Validation of therapeutic approaches targeting epigenetic enzymes in a pre-clinical model of congenital myopathies. Research Project | 2 Project MembersSkeletal muscle contraction starts when Ca2+ is released from the sarcoplasmic reticulum via the ryanodine receptor (RyR1). Defects in Ca2+ signaling, often due to RYR1 mutations, cause neuromuscular disorders like malignant hyperthermia and congenital myopathies. To study recessive RYR1 mutations, a mouse model (dHT) was created with specific mutations found in a child with multi-minicore disease (MmD). Treatment of dHT mice with drugs targeting epigenetic enzymes (HDACs and DNMT inhibitors) improved muscle strength and function. The study aims to explore the biochemical changes these drugs trigger, potentially leading to clinical trials for treating congenital myopathies. Air Pollution and Effects on Lung Functional Development and Respiratory Morbidity in At-Risk Infants Research Project | 4 Project MembersBACKGROUND AND RATIONALE: This is a direct continuation of SNF 182871/1, which investigated the impact of early-childhood environmental factors on lung functional growth and consequences for later respiratory morbidity in healthy term infants. We previously demonstrated that even low-level air pollution exposure during pregnancy and early childhood is associated with impaired lung functional growth in infancy and early childhood. Although the mechanisms are still unclear, they could be related to lung functional growth deficits or remodeling of the lung due to changes in the intrauterine environment. Air pollution is known to induce oxidative stress response and related autophagy and cellular senescence mechanisms, potentially playing a role in pollution-related lung pathology and in remodeling. As novel preliminary evidence in SNF 182871/1, we recently found that, in the cord blood of human infants, autophagy-related biomarkers are correlated with remodeling biomarkers. We also found that air pollution exposure during pregnancy is associated with biomarkers of autophagy and remodeling in the cord blood of healthy term infants. Interestingly, these mechanisms also play an important role in fetal development and preterm birth, and may thus theoretically contribute to the susceptibility of infants-and particularly preterm infants-to oxidative stress and air pollution effects. Indeed, as first evidence from SNF 182871/1, we also found an enhanced impact of air pollutants on lung function impairment of preterm infants. Furthermore, our own preliminary human data show that markers of autophagy, and remodeling already have significant differences between the cord blood of preterm infants compared to term infants at birth prior to early postnatal injury. Bringing this together, we hypothesize that the interaction of oxidative stress response, autophagy and remodeling could be a key mechanism involved in the complex host-environment interaction determining lung functional growth and related respiratory morbidity. Moreover, this response could be different in infants at risk for chronic respiratory symptoms, such preterm infants, infants born from asthmatic mothers or infants exposed to high levels of air pollution during pregnancy. OVERALL OBJECTIVES: We aim to expand the ongoing BILD cohort of (i) term infants with two risk subgroups, (ii) infants born preterm, and (iii) infants born to asthmatic mothers, and we will investigate the differences in response to prenatal air pollution in relation to the above key mechanisms. SPECIFIC AIMS: In comparison to healthy term infants, we will investigate in study phase 1, (i) whether the increased susceptibility of infants to prenatal air pollution in these three risk groups is related to differences in markers of oxidative stress response, autophagy, and remodeling in cord blood and in study phase 2, (ii) whether these pollution-related cord blood profiles are correlated to lung functional development and subsequent symptoms in the first year of life (primary outcomes) and at school age (secondary outcomes). We will replicate these findings in other birth cohorts from collaborators (Germany, Australia) with comparable outcome measures. METHODS: In our prospective BILD birth cohort of 1000 unselected healthy term infants, 400 preterm infants, and 200 infants from asthmatic mothers we will (i) estimate indoor and outdoor air pollution exposure during pregnancy and in early infancy, (ii) assess family, obstetric and birth history, cord-molecular biomarkers (metabolomics, gene expression, proteins), and infant lung function shortly after birth (including exhalomics) and at 6 years of age, as well as respiratory symptoms in the first year of life and at school age. EXPECTED RESULTS AND IMPACT: We expect a 26.03.2021 18:35:26 Page - 14 - significant correlation between air pollution exposure and oxidative stress response and lung remodeling in newborns with effects on lung function and clinical outcomes, the latter effects enhanced in the risk groups. Particularly for these risk groups, today's air pollution may already result in lung remodeling and subsequent impaired lung functional growth even at this early stage of life. Since early-life lung functional impairment often persists until school age and even late adulthood, it is a previously described early-life risk factor known to be associated with asthma in children and chronic obstructive respiratory airway diseases in the elderly. Thus, early-life environmental injury has a potentially very relevant impact on future global respiratory health, with unpredictable costs. We are one of the first groups to look into the impact of these air-pollution-induced mechanisms on oxidative stress response and lung remodeling, subsequent impairment of lung functional growth, and resulting human lung disease. Better understanding of these mechanisms might help the development of preventative and therapeutic strategies, particularly for at-risk infants. Impact of mobile communication signals on the regulation of neural differentiation Research Project | 4 Project MembersOur project aims at studying possible effects of radiofrequency electromagnetic fields (RF-EMF) on neuronal differentiation and related cellular pathways known to be involved in neurodegeneration and associated diseases. Focusing on pathways playing a role in neuronal differentiation and degeneration, we will investigate RF-EMF effects on distinct neuronal cell populations before and during differentiation and identify molecular pathways involved in a hypothesis-free genomic approach as well as in a hypothesis-driven approach. These goals will be achieved by applying cell culture-based experimentations including neuron-like and neuronal stem/progenitor cells. Progression of differentiation and phenotypic characterization will be assessed by fluorescence microscopy of stem cell and neuronal markers combined with high-content analysis and evaluation of morphology (i.e. neurite outgrowth). These analyses will be complemented by the assessment of key players of cellular pathways (e.g. the ERK/MAP-K, PI3-K/Akt, Wnt/β-catenin) underlying the morphological changes such as neurite outgrowth in a quantitative manner (e.g. by Western blotting and FRET-biosensor). Mitochondrial dysfunction during neurodegeneration was shown to increase the production of reactive oxygen species (ROS). Oxidative stress is a frequent early condition in the pathogenesis of neurodegenerative diseases, is often observed in cells upon EMF exposure and has an evident potential to compromise genetic as well as epigenetic stability. Mitochondrial integrity, an important indicator of neuronal aging and degeneration will be investigated quantitatively. To assess the impact of RF-EMF exposure on the composition and behavior of the differentiating neural cell population, gene expression profiling (transcriptome) of single cells before and after differentiation will be performed. Cluster analysis of gene expression profiles will describe the dynamics of the cell population and potentially result in the identification of target regions with altered epigenetic modifications. Target-specific quantitation of DNA methylation and histone modifications at regulatory elements (promoter, enhancer) of identified genes, markers of neural stem cell and the neuronal lineage will be analyzed by pyrosequencing of bisulfite-converted DNA and by chromatin immunoprecipitation (ChIP). This project will provide a significant and critical insight into the adverse effects of exposure to modulated RF-EMF as used for mobile communication (GSM) on signaling cascades and physiology as well as on morphological and epigenetic characteristics of neural cells in vitro . BERENIS Berater Research Project | 1 Project MembersDas BAFU hat als Umweltfachstelle des Bundes die Aufgabe, die Forschung über gesundheitliche Auswirkungen nichtionisierender Strahlung von Infrastrukturanlagen wie auch diesbezügliche Alltagserfahrungen zu verfolgen und die Ergebnisse zu bewerten, sowie die Öffentlichkeit über den Stand der Wissenschaft und der Erfahrung zu informieren. Sofern neue Ergebnisse dies erfordern, stellt es dem Bundesrat Antrag auf eine Anpassung der Immissionsgrenzwerte der NISV. Andernfalls muss es - insbesondere gegenüber technologiekritischen Kreisen - jederzeit begründen können, weshalb die geltenden Immissionsgrenzwerte nicht angepasst werden müssen. Die Bewertung muss Aussagen darüber machen, wie sicher biologische Effekte nachgewiesen sind, ob sie für die Gesundheit relevant sind und wie viele Menschen gegebenenfalls betroffen sind. Zur Bewertung gehört auch eine Früherkennung potenzieller Risiken. Dem Bundesrat wird in gerichtlichen Beschwerdeverfahren regelmässig vorgeworfen, die Grenzwerte der NISV nicht nach dem Stand der Wissenschaft oder der Erfahrung festgesetzt zu haben. Beschwerdeführer legen eigene Bewertungen des Standes der Wissenschaft vor, um wesentlich strengere Grenzwerte zu fordern. Das Bundesgericht hat diese Begehren bisher mit dem Hinweis zurückgewiesen, dass es Aufgabe der Fachbehörden des Bundes sei, den Stand der Wissenschaft zu evaluieren, und dass das BAFU dieser Aufgabe nachkomme. Die zuständige Sektion NIS des BAFU hat bisher das Schweiz. Tropen- und Public Health Institut Basel (Swiss TPH) mit der Bewertungsarbeit beauftragt, und kurze Zeit auch das Institut für Biomedizin der Universität Basel. Das TPH hat die wissenschaftlichen Publikationen zu Humanstudien möglichst vollständig erfasst, bewertet und in der Datenbank ELMAR öffentlich zugänglich gemacht. Daraus hat es mehrere thematische Syntheseberichte erstellt, welche das BAFU in seinen Publikationsreihen publiziert hat. Diese Bewertungsarbeit konzentrierte sich auf Humanstudien. Der Fokus soll nun auf experimentelle (Tiere und Zellen) sowie mechanistische Studien ausgeweitet und die Verantwortung für die Bewertung breiter abgestützt werden, um eine umfassendere Sicht des wissenschaftlichen Kenntnisstandes zu erhalten. Zu diesem Zweck wird eine beratende Expertengruppe eingesetzt. Analysis of TDG Dependent Ribonucloprotein Complex formation on Long Noncoding RNA by SILAC Research Project | 1 Project MembersThe generation of a diversity of cells with different functions is crucial for multicellular organisms. The individual cell identity relies on a selective readout of the genome achieved with the help of epigenetic mechanisms which direct the transcriptional programs to cell-type specific endpoints. Working at the level of chromatin, these mechanisms orchestrate gene transcription by deposition, exchange or removal of epigenetic modifications at histones as wells as in the DNA. Inaccuracy of the epigenetic system not only impedes successful establishment of cell identity but may also have deleterious effects on tissue homeostasis. This is best exemplified by epigenetically unstable cancers which frequently show aberrant patterns of CpG methylation. Thus, a faithful control of the epigenetic machinery is critical for the preservation of correct cell identity. We have identified Thymine DNA glycosylase as a potential epigenetic proof-reader in CpG rich promoters of genes in active and bivalent chromatin. TDG helps to maintain these regions in an unmethylated state by initiating a DNA repair process at sites of erroneous cytosine methylation. Research of the last years has shed light on how TDG dependent active demethylation may occur at the molecular level; yet, the ways how this process is regulated and targeted to its site of actions remain obscure. Hints on how this may be achieved come from knowledge obtained for long noncoding RNAs. Serving as scaffolds, some of these RNAs have been shown to play a central role for the regulation of epigenetic processes. Together with our own preliminary data showing association of TDG with a variety of lncRNAs this suggests that these RNAs may be important for TDG mediated active DNA demethylation. Intriguingly, previously identified interaction partners of TDG, such as e.g. nuclear receptors have been reported to associate with some of the identified TDG bound lncRNAs, especially steroid receptor RNA activator 1. The work proposed herein will follow-up on this important question and investigate the TDG dependent composition of ribonucleoprotein complex by SILAC using SRA1 as a model lncRNA. Sub-cellular targeting microscopy - Signaling in Development and Oncology Research Project | 6 Project MembersIn the past decade, research in development and oncology has uncovered an impressive number of relevant signaling pathways. While a qualitative understanding of signals, and their connection to cellular outputs has been established, plasticity and feedback of signaling networks remain obscured. A better understanding of dynamic and locally constrained signaling events driving organ development and disease progression requires access to refined subcellular probe detection. The availability of optogenetic and chemical biology tools provides novel opportunities, but requires dedicated microscopy equiment. For this reason, six projects at the Department of Biomedicine (DBM) and the Biocenter (BC) of the Univerisity of Basel i) localized lipid signaling in disease (M. Wymann, DBM); ii) dynamic subcellular Wnt/b-catenin signaling in epithelial mesenchymal transition (G. Christofori, DBM); iii) DNA dynamics and confined epigenetic plasticity (P. Schär, DBM); iv) real-time monitoring of Sonic Hedgehog and Bone Morphogenetic Protein gradients in limb buds (R. Zeller, DBM); v) ultrastructural analysis of neuronal stem cell control (V. Taylor, DBM); and vi) molecular mechanisms determining the development of vascular networks (M. Affolter, BC), illustrate the need of the requested "subcellular targeting microscopy" equipment. The core of the platform is a highly sensitive microlens-enhanced spinning disk microscope linked to FRAP, ablation, and multiple excitation laser lines, and an integrated TIRF module to monitor plasma membrane events. Through its integration into the BioOptics core facility at the DBM, the subcellular targeting platform will be accessible to >500 regional researchers at the University of Basel, DBM, FMI, D-BSSE, Fachhochschule, etc. An image storage and analysis pipeline with remote user access capabilitiy is in place, to allow seamless operation and output. The requested equipment will allow the use of genetically encoded opto-genetic proteins, proteins tagged for reactivity with chemical inducers of dimerisation (CIDs), and the possibility to perform FRAP/TIRF and FRAP/confocal microscopy, and will greatly enhance the possibilities to manipulate and track subcellular localization of target proteins. The insights gained by these experimental approaches will be critical for a better understanding of dynamic biological processes, and will spur the design of innovative therapeutic approaches to counteract resistance mechanisms in oncology. Characterization of the Ewing's sarcoma protein's involvement in the maintenance of genomic stability Research Project | 3 Project MembersConsidering that a variety of genetic abnormalities typically occur in a given tumor, it is striking that some cancers exhibit highly characteristic genomic changes. They consist in chromosome fusions that occur at very specific chromosomal locations, one of which is always the locus of a FET protein. FET stands for the family of fused in sarcoma (FUS), Ewing sarcoma (EWS) and TATA box binding protein associated factor 15 (TAF15) proteins. The most common fusion proteins are EWS-FLI1 in Ewing sarcoma, FUS-CREB3L1 in low-grade fibromyxoid sarcoma and TAF15-NR4A3 in extraskeletal myxoid chondrosarcoma. Although normal FET proteins bind RNA, this activity is absent in the fusion proteins. Whether this loss is relevant for the development of the tumor is not known. In a previous study we mapped the RNA targets of EWS. To our surprise, we found that EWS binds RNAs that come from genomic loci that are prone to 'genomic instability'. We further obtained evidence that EWS helps the transcription of DNA into messenger RNAs, facilitating the progression of the RNA polymerization enzyme (RNAPII) through loci that are prone to form stable structures that impede transcription. In this project we would like to characterize the relationship between EWS and RNAPII-dependent transcription, and the consequences at the cellular level of the long-term reduction in EWS expression. We will employ cells and cell lines in which we can reduce EWS expression in a controlled manner. We will determine the rate at which the RNA polymerase II elongates transcripts genome-wide and the frequency with which lesions in the DNA that are known as DNA double-strand breaks occur when EWS is limiting. Furthermore, we will investigate the long-term consequences of reduced EWS expression on cell morphology, viability and functionality. Our study could thus contribute to an improved understanding of the pathogenic mechanisms of Ewing's sarcoma. Cellular and molecular effects of pulsed electromagnetic fields Research Project | 3 Project MembersInterfering with cell proliferation (growth control) is an established and successful concept of both, conventional and targeted cancer therapy, but often the therapeutic choice is a double edged sword. Treatments interfering with cell proliferation are potentially harmful because they are severely toxic and often cause undesirable short and long side-effects on different tissues, adversely affecting the patient's well-being. In this respect, the observation that low intensity pulsed electromagnetic fields (PEMFs) interfere with the proliferation of cancer cells may reflect a promising strategy for future cancer therapeutics. Importantly, non-cancerous tissue cells do not seem to be affected and stem cells respond by increased proliferation and and/or differentiation. Molecular targets and mechanisms underlying this potential therapeutic effect of PEMFs are poorly understood to date, but there are indications pointing to an involvement of signalling cascades, mitotic failure and induced apoptosis. The investigation and characterization of these mechanisms at the cellular and molecular level is the focus of the proposed project. It involves cell culture-based experimental approaches using advanced microscopy and molecular biological means to explore the PEMF-induced network of anti-proliferative signals and response of cancer cells from different origin in comparison to differentiated and undifferentiated cells. Key readouts will be parameters of cell proliferation (i.e. cell viability, proliferation and apoptosis), mitotic and cell phase progression and growth-, stress- and checkpoint signalling activities. The proposed experiments aim to reveal whether PEMF-mediated reduction of cell proliferation is a common phenomenon of cancer cells or rather restricted to a spectrum of responsive cancers. Designed to identify the responsive targets, pathways and structures, they will extend the current understanding about how cells react instantly to PEMF exposure and how these responses are then transduced to produce the cellular consequences observed. The proposed work will thus provide insight into how PEMFs and possibly other types of electromagnetic signals interact with cellular processes to disturb cell proliferation and, thereby, a mechanistic framework for a rational design of therapeutic applications. DNA Repair, Epigenetic Stability, and CpG Island Hypermethylation in Colorectal Tumorigenesis Research Project | 1 Project MembersNo Description available Impact of DNA ligase IV on genome stability during DNA replication Research Project | 1 Project MembersResearch grant to Dr. Olivier Fritsch 12 12
Validation of therapeutic approaches targeting epigenetic enzymes in a pre-clinical model of congenital myopathies. Research Project | 2 Project MembersSkeletal muscle contraction starts when Ca2+ is released from the sarcoplasmic reticulum via the ryanodine receptor (RyR1). Defects in Ca2+ signaling, often due to RYR1 mutations, cause neuromuscular disorders like malignant hyperthermia and congenital myopathies. To study recessive RYR1 mutations, a mouse model (dHT) was created with specific mutations found in a child with multi-minicore disease (MmD). Treatment of dHT mice with drugs targeting epigenetic enzymes (HDACs and DNMT inhibitors) improved muscle strength and function. The study aims to explore the biochemical changes these drugs trigger, potentially leading to clinical trials for treating congenital myopathies.
Air Pollution and Effects on Lung Functional Development and Respiratory Morbidity in At-Risk Infants Research Project | 4 Project MembersBACKGROUND AND RATIONALE: This is a direct continuation of SNF 182871/1, which investigated the impact of early-childhood environmental factors on lung functional growth and consequences for later respiratory morbidity in healthy term infants. We previously demonstrated that even low-level air pollution exposure during pregnancy and early childhood is associated with impaired lung functional growth in infancy and early childhood. Although the mechanisms are still unclear, they could be related to lung functional growth deficits or remodeling of the lung due to changes in the intrauterine environment. Air pollution is known to induce oxidative stress response and related autophagy and cellular senescence mechanisms, potentially playing a role in pollution-related lung pathology and in remodeling. As novel preliminary evidence in SNF 182871/1, we recently found that, in the cord blood of human infants, autophagy-related biomarkers are correlated with remodeling biomarkers. We also found that air pollution exposure during pregnancy is associated with biomarkers of autophagy and remodeling in the cord blood of healthy term infants. Interestingly, these mechanisms also play an important role in fetal development and preterm birth, and may thus theoretically contribute to the susceptibility of infants-and particularly preterm infants-to oxidative stress and air pollution effects. Indeed, as first evidence from SNF 182871/1, we also found an enhanced impact of air pollutants on lung function impairment of preterm infants. Furthermore, our own preliminary human data show that markers of autophagy, and remodeling already have significant differences between the cord blood of preterm infants compared to term infants at birth prior to early postnatal injury. Bringing this together, we hypothesize that the interaction of oxidative stress response, autophagy and remodeling could be a key mechanism involved in the complex host-environment interaction determining lung functional growth and related respiratory morbidity. Moreover, this response could be different in infants at risk for chronic respiratory symptoms, such preterm infants, infants born from asthmatic mothers or infants exposed to high levels of air pollution during pregnancy. OVERALL OBJECTIVES: We aim to expand the ongoing BILD cohort of (i) term infants with two risk subgroups, (ii) infants born preterm, and (iii) infants born to asthmatic mothers, and we will investigate the differences in response to prenatal air pollution in relation to the above key mechanisms. SPECIFIC AIMS: In comparison to healthy term infants, we will investigate in study phase 1, (i) whether the increased susceptibility of infants to prenatal air pollution in these three risk groups is related to differences in markers of oxidative stress response, autophagy, and remodeling in cord blood and in study phase 2, (ii) whether these pollution-related cord blood profiles are correlated to lung functional development and subsequent symptoms in the first year of life (primary outcomes) and at school age (secondary outcomes). We will replicate these findings in other birth cohorts from collaborators (Germany, Australia) with comparable outcome measures. METHODS: In our prospective BILD birth cohort of 1000 unselected healthy term infants, 400 preterm infants, and 200 infants from asthmatic mothers we will (i) estimate indoor and outdoor air pollution exposure during pregnancy and in early infancy, (ii) assess family, obstetric and birth history, cord-molecular biomarkers (metabolomics, gene expression, proteins), and infant lung function shortly after birth (including exhalomics) and at 6 years of age, as well as respiratory symptoms in the first year of life and at school age. EXPECTED RESULTS AND IMPACT: We expect a 26.03.2021 18:35:26 Page - 14 - significant correlation between air pollution exposure and oxidative stress response and lung remodeling in newborns with effects on lung function and clinical outcomes, the latter effects enhanced in the risk groups. Particularly for these risk groups, today's air pollution may already result in lung remodeling and subsequent impaired lung functional growth even at this early stage of life. Since early-life lung functional impairment often persists until school age and even late adulthood, it is a previously described early-life risk factor known to be associated with asthma in children and chronic obstructive respiratory airway diseases in the elderly. Thus, early-life environmental injury has a potentially very relevant impact on future global respiratory health, with unpredictable costs. We are one of the first groups to look into the impact of these air-pollution-induced mechanisms on oxidative stress response and lung remodeling, subsequent impairment of lung functional growth, and resulting human lung disease. Better understanding of these mechanisms might help the development of preventative and therapeutic strategies, particularly for at-risk infants.
Impact of mobile communication signals on the regulation of neural differentiation Research Project | 4 Project MembersOur project aims at studying possible effects of radiofrequency electromagnetic fields (RF-EMF) on neuronal differentiation and related cellular pathways known to be involved in neurodegeneration and associated diseases. Focusing on pathways playing a role in neuronal differentiation and degeneration, we will investigate RF-EMF effects on distinct neuronal cell populations before and during differentiation and identify molecular pathways involved in a hypothesis-free genomic approach as well as in a hypothesis-driven approach. These goals will be achieved by applying cell culture-based experimentations including neuron-like and neuronal stem/progenitor cells. Progression of differentiation and phenotypic characterization will be assessed by fluorescence microscopy of stem cell and neuronal markers combined with high-content analysis and evaluation of morphology (i.e. neurite outgrowth). These analyses will be complemented by the assessment of key players of cellular pathways (e.g. the ERK/MAP-K, PI3-K/Akt, Wnt/β-catenin) underlying the morphological changes such as neurite outgrowth in a quantitative manner (e.g. by Western blotting and FRET-biosensor). Mitochondrial dysfunction during neurodegeneration was shown to increase the production of reactive oxygen species (ROS). Oxidative stress is a frequent early condition in the pathogenesis of neurodegenerative diseases, is often observed in cells upon EMF exposure and has an evident potential to compromise genetic as well as epigenetic stability. Mitochondrial integrity, an important indicator of neuronal aging and degeneration will be investigated quantitatively. To assess the impact of RF-EMF exposure on the composition and behavior of the differentiating neural cell population, gene expression profiling (transcriptome) of single cells before and after differentiation will be performed. Cluster analysis of gene expression profiles will describe the dynamics of the cell population and potentially result in the identification of target regions with altered epigenetic modifications. Target-specific quantitation of DNA methylation and histone modifications at regulatory elements (promoter, enhancer) of identified genes, markers of neural stem cell and the neuronal lineage will be analyzed by pyrosequencing of bisulfite-converted DNA and by chromatin immunoprecipitation (ChIP). This project will provide a significant and critical insight into the adverse effects of exposure to modulated RF-EMF as used for mobile communication (GSM) on signaling cascades and physiology as well as on morphological and epigenetic characteristics of neural cells in vitro .
BERENIS Berater Research Project | 1 Project MembersDas BAFU hat als Umweltfachstelle des Bundes die Aufgabe, die Forschung über gesundheitliche Auswirkungen nichtionisierender Strahlung von Infrastrukturanlagen wie auch diesbezügliche Alltagserfahrungen zu verfolgen und die Ergebnisse zu bewerten, sowie die Öffentlichkeit über den Stand der Wissenschaft und der Erfahrung zu informieren. Sofern neue Ergebnisse dies erfordern, stellt es dem Bundesrat Antrag auf eine Anpassung der Immissionsgrenzwerte der NISV. Andernfalls muss es - insbesondere gegenüber technologiekritischen Kreisen - jederzeit begründen können, weshalb die geltenden Immissionsgrenzwerte nicht angepasst werden müssen. Die Bewertung muss Aussagen darüber machen, wie sicher biologische Effekte nachgewiesen sind, ob sie für die Gesundheit relevant sind und wie viele Menschen gegebenenfalls betroffen sind. Zur Bewertung gehört auch eine Früherkennung potenzieller Risiken. Dem Bundesrat wird in gerichtlichen Beschwerdeverfahren regelmässig vorgeworfen, die Grenzwerte der NISV nicht nach dem Stand der Wissenschaft oder der Erfahrung festgesetzt zu haben. Beschwerdeführer legen eigene Bewertungen des Standes der Wissenschaft vor, um wesentlich strengere Grenzwerte zu fordern. Das Bundesgericht hat diese Begehren bisher mit dem Hinweis zurückgewiesen, dass es Aufgabe der Fachbehörden des Bundes sei, den Stand der Wissenschaft zu evaluieren, und dass das BAFU dieser Aufgabe nachkomme. Die zuständige Sektion NIS des BAFU hat bisher das Schweiz. Tropen- und Public Health Institut Basel (Swiss TPH) mit der Bewertungsarbeit beauftragt, und kurze Zeit auch das Institut für Biomedizin der Universität Basel. Das TPH hat die wissenschaftlichen Publikationen zu Humanstudien möglichst vollständig erfasst, bewertet und in der Datenbank ELMAR öffentlich zugänglich gemacht. Daraus hat es mehrere thematische Syntheseberichte erstellt, welche das BAFU in seinen Publikationsreihen publiziert hat. Diese Bewertungsarbeit konzentrierte sich auf Humanstudien. Der Fokus soll nun auf experimentelle (Tiere und Zellen) sowie mechanistische Studien ausgeweitet und die Verantwortung für die Bewertung breiter abgestützt werden, um eine umfassendere Sicht des wissenschaftlichen Kenntnisstandes zu erhalten. Zu diesem Zweck wird eine beratende Expertengruppe eingesetzt.
Analysis of TDG Dependent Ribonucloprotein Complex formation on Long Noncoding RNA by SILAC Research Project | 1 Project MembersThe generation of a diversity of cells with different functions is crucial for multicellular organisms. The individual cell identity relies on a selective readout of the genome achieved with the help of epigenetic mechanisms which direct the transcriptional programs to cell-type specific endpoints. Working at the level of chromatin, these mechanisms orchestrate gene transcription by deposition, exchange or removal of epigenetic modifications at histones as wells as in the DNA. Inaccuracy of the epigenetic system not only impedes successful establishment of cell identity but may also have deleterious effects on tissue homeostasis. This is best exemplified by epigenetically unstable cancers which frequently show aberrant patterns of CpG methylation. Thus, a faithful control of the epigenetic machinery is critical for the preservation of correct cell identity. We have identified Thymine DNA glycosylase as a potential epigenetic proof-reader in CpG rich promoters of genes in active and bivalent chromatin. TDG helps to maintain these regions in an unmethylated state by initiating a DNA repair process at sites of erroneous cytosine methylation. Research of the last years has shed light on how TDG dependent active demethylation may occur at the molecular level; yet, the ways how this process is regulated and targeted to its site of actions remain obscure. Hints on how this may be achieved come from knowledge obtained for long noncoding RNAs. Serving as scaffolds, some of these RNAs have been shown to play a central role for the regulation of epigenetic processes. Together with our own preliminary data showing association of TDG with a variety of lncRNAs this suggests that these RNAs may be important for TDG mediated active DNA demethylation. Intriguingly, previously identified interaction partners of TDG, such as e.g. nuclear receptors have been reported to associate with some of the identified TDG bound lncRNAs, especially steroid receptor RNA activator 1. The work proposed herein will follow-up on this important question and investigate the TDG dependent composition of ribonucleoprotein complex by SILAC using SRA1 as a model lncRNA.
Sub-cellular targeting microscopy - Signaling in Development and Oncology Research Project | 6 Project MembersIn the past decade, research in development and oncology has uncovered an impressive number of relevant signaling pathways. While a qualitative understanding of signals, and their connection to cellular outputs has been established, plasticity and feedback of signaling networks remain obscured. A better understanding of dynamic and locally constrained signaling events driving organ development and disease progression requires access to refined subcellular probe detection. The availability of optogenetic and chemical biology tools provides novel opportunities, but requires dedicated microscopy equiment. For this reason, six projects at the Department of Biomedicine (DBM) and the Biocenter (BC) of the Univerisity of Basel i) localized lipid signaling in disease (M. Wymann, DBM); ii) dynamic subcellular Wnt/b-catenin signaling in epithelial mesenchymal transition (G. Christofori, DBM); iii) DNA dynamics and confined epigenetic plasticity (P. Schär, DBM); iv) real-time monitoring of Sonic Hedgehog and Bone Morphogenetic Protein gradients in limb buds (R. Zeller, DBM); v) ultrastructural analysis of neuronal stem cell control (V. Taylor, DBM); and vi) molecular mechanisms determining the development of vascular networks (M. Affolter, BC), illustrate the need of the requested "subcellular targeting microscopy" equipment. The core of the platform is a highly sensitive microlens-enhanced spinning disk microscope linked to FRAP, ablation, and multiple excitation laser lines, and an integrated TIRF module to monitor plasma membrane events. Through its integration into the BioOptics core facility at the DBM, the subcellular targeting platform will be accessible to >500 regional researchers at the University of Basel, DBM, FMI, D-BSSE, Fachhochschule, etc. An image storage and analysis pipeline with remote user access capabilitiy is in place, to allow seamless operation and output. The requested equipment will allow the use of genetically encoded opto-genetic proteins, proteins tagged for reactivity with chemical inducers of dimerisation (CIDs), and the possibility to perform FRAP/TIRF and FRAP/confocal microscopy, and will greatly enhance the possibilities to manipulate and track subcellular localization of target proteins. The insights gained by these experimental approaches will be critical for a better understanding of dynamic biological processes, and will spur the design of innovative therapeutic approaches to counteract resistance mechanisms in oncology.
Characterization of the Ewing's sarcoma protein's involvement in the maintenance of genomic stability Research Project | 3 Project MembersConsidering that a variety of genetic abnormalities typically occur in a given tumor, it is striking that some cancers exhibit highly characteristic genomic changes. They consist in chromosome fusions that occur at very specific chromosomal locations, one of which is always the locus of a FET protein. FET stands for the family of fused in sarcoma (FUS), Ewing sarcoma (EWS) and TATA box binding protein associated factor 15 (TAF15) proteins. The most common fusion proteins are EWS-FLI1 in Ewing sarcoma, FUS-CREB3L1 in low-grade fibromyxoid sarcoma and TAF15-NR4A3 in extraskeletal myxoid chondrosarcoma. Although normal FET proteins bind RNA, this activity is absent in the fusion proteins. Whether this loss is relevant for the development of the tumor is not known. In a previous study we mapped the RNA targets of EWS. To our surprise, we found that EWS binds RNAs that come from genomic loci that are prone to 'genomic instability'. We further obtained evidence that EWS helps the transcription of DNA into messenger RNAs, facilitating the progression of the RNA polymerization enzyme (RNAPII) through loci that are prone to form stable structures that impede transcription. In this project we would like to characterize the relationship between EWS and RNAPII-dependent transcription, and the consequences at the cellular level of the long-term reduction in EWS expression. We will employ cells and cell lines in which we can reduce EWS expression in a controlled manner. We will determine the rate at which the RNA polymerase II elongates transcripts genome-wide and the frequency with which lesions in the DNA that are known as DNA double-strand breaks occur when EWS is limiting. Furthermore, we will investigate the long-term consequences of reduced EWS expression on cell morphology, viability and functionality. Our study could thus contribute to an improved understanding of the pathogenic mechanisms of Ewing's sarcoma.
Cellular and molecular effects of pulsed electromagnetic fields Research Project | 3 Project MembersInterfering with cell proliferation (growth control) is an established and successful concept of both, conventional and targeted cancer therapy, but often the therapeutic choice is a double edged sword. Treatments interfering with cell proliferation are potentially harmful because they are severely toxic and often cause undesirable short and long side-effects on different tissues, adversely affecting the patient's well-being. In this respect, the observation that low intensity pulsed electromagnetic fields (PEMFs) interfere with the proliferation of cancer cells may reflect a promising strategy for future cancer therapeutics. Importantly, non-cancerous tissue cells do not seem to be affected and stem cells respond by increased proliferation and and/or differentiation. Molecular targets and mechanisms underlying this potential therapeutic effect of PEMFs are poorly understood to date, but there are indications pointing to an involvement of signalling cascades, mitotic failure and induced apoptosis. The investigation and characterization of these mechanisms at the cellular and molecular level is the focus of the proposed project. It involves cell culture-based experimental approaches using advanced microscopy and molecular biological means to explore the PEMF-induced network of anti-proliferative signals and response of cancer cells from different origin in comparison to differentiated and undifferentiated cells. Key readouts will be parameters of cell proliferation (i.e. cell viability, proliferation and apoptosis), mitotic and cell phase progression and growth-, stress- and checkpoint signalling activities. The proposed experiments aim to reveal whether PEMF-mediated reduction of cell proliferation is a common phenomenon of cancer cells or rather restricted to a spectrum of responsive cancers. Designed to identify the responsive targets, pathways and structures, they will extend the current understanding about how cells react instantly to PEMF exposure and how these responses are then transduced to produce the cellular consequences observed. The proposed work will thus provide insight into how PEMFs and possibly other types of electromagnetic signals interact with cellular processes to disturb cell proliferation and, thereby, a mechanistic framework for a rational design of therapeutic applications.
DNA Repair, Epigenetic Stability, and CpG Island Hypermethylation in Colorectal Tumorigenesis Research Project | 1 Project MembersNo Description available
Impact of DNA ligase IV on genome stability during DNA replication Research Project | 1 Project MembersResearch grant to Dr. Olivier Fritsch
