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Institute
Trojanowski, Peter ; Plötner, Jürgen ; Grünewald, Christian ; Braun, Markus ; Reuß, Andreas ; Engels, Joachim W. ; Wachtveitl, Josef
We examine the photoinduced excited state dynamics of pyrene modified adenosine, a versatile probe for folding and hybridization of ribonucleic acids. Measurements in different solvents revealed complex ultrafast dynamics, but high robustness since the overall fluorescence quantum yield (Φf) is hardly affected. The result is a strong fluorescent RNA-probe whose spectral properties change in a defined way upon environmental changes.
Zur prototypischen Modellierung ausgewählter Völker im Tabletop-Strategiespiel Warhammer Fantasy
(2013)
Der vorliegende Beitrag beschäftigt sich mit der Typisierung einzelner Völker des Tabletop-Strategiespiels 'Warhammer Fantasy'. Unter methodischen Gesichtspunkten kann man für die Ermittlung von Prototypen auf bewährte textlinguistische bzw. lexikalisch-semantische Verfahrensweisen zurückgreifen. Im Folgenden sollen Referenzidentitäten, Isotopieketten, Attribuierungen und Konnotationen betrachtet werden. Die Fülle des vorhandenen Textmaterials (siehe dazu Abschnitt 3) macht eine Fokussierung unumgänglich. Vorliegende Studie beschränkt sich auf die Textgruppe der völkerspezifischen Armeebücher. Besondere Aufmerksamkeit erfahren hierbei einerseits der Text auf dem Rückendeckel (U4), schlichtweg deshalb, weil die Vermutung naheliegt, dass diese Stelle für eine prototypische Verdichtung besonders exponiert ist, und andererseits die Bezeichnungen und Beschreibungen der armeetypischen Einheitentypen, welche die völkerspezifischen Binnenstrukturierungen aufzeigen.
Fusoselect : cell-cell fusion activity engineered by directed evolution of a retroviral glycoprotein
(2006)
Merten, Christoph A. ; Stitz, Jörn ; Braun, Gundula ; Medvedovska, Julia ; Cichutek, Klaus ; Buchholz, Christian
Membrane fusion plays a key role in many biological processes including vesicle trafficking, synaptic transmission, fertilization or cell entry of enveloped viruses. As a common feature the fusion process is mediated by distinct membrane proteins. We describe here "Fusoselect", a universal procedure allowing the identification and engineering of molecular determinants for cell–cell fusion-activity by directed evolution. The system couples cell–cell fusion with the release of retroviral particles, but can principally be applied to membrane proteins of non-viral origin as well. As a model system, we chose a γ-retroviral envelope protein, which naturally becomes fusion-active through proteolytic processing by the viral protease. The selection process evolved variants that, in contrast to the parental protein, mediated cell–cell fusion in absence of the viral protease. Detailed analysis of the variants revealed molecular determinants for fusion competence in the cytoplasmic tail (CT) of retroviral Env proteins and demonstrated the power of Fusoselect.
Scheidt, Moritz von ; Bauer, Sabine ; Ma, Angela ; Hao, Ke ; Kessler, Thorsten ; Vilne, Baiba ; Wang, Ying ; Hodonsky, Chani J. ; Ghosh, Saikat K.B. ; Mokry, Michal ; Gao, Hua ; Kawai, Kenji ; Sakamoto, Atsushi ; Kaiser, Juliane ; Bongiovanni, Dario ; Fleig, Julia ; Oldenbuettel, Lilith ; Chen, Zhifen ; Moggio, Aldo ; Sager, Hendrik ; Hecker, Judith S. ; Bassermann, Florian ; Mägdefessel, Lars ; Miller, Clint L. ; Koenig, Wolfgang ; Zeiher, Andreas M. ; Dimmeler, Stefanie ; Graw, Matthias ; Braun, Christian ; Ruusalepp, Arno ; Leeper, Nicholas J. ; Kovacic, Jason C. ; Björkegren, Johan L.M. ; Schunkert, Heribert
Background: Leukocyte progenitors derived from clonal hematopoiesis of undetermined potential (CHIP) are associated with increased cardiovascular events. However, the prevalence and functional relevance of CHIP in coronary artery disease (CAD) are unclear, and cells affected by CHIP have not been detected in human atherosclerotic plaques.
Methods: CHIP mutations in blood and tissues were identified by targeted deep-DNA-sequencing (DNAseq: coverage >3,000) and whole-genome-sequencing (WGS: coverage >35). CHIP-mutated leukocytes were visualized in human atherosclerotic plaques by mutaFISHTM. Functional relevance of CHIP mutations was studied by RNAseq.
Results: DNAseq of whole blood from 540 deceased CAD patients of the Munich cardIovaScular StudIes biObaNk (MISSION) identified 253 (46.9%) CHIP mutation carriers (mean age 78.3 years). DNAseq on myocardium, atherosclerotic coronary and carotid arteries detected identical CHIP mutations in 18 out of 25 mutation carriers in tissue DNA. MutaFISHTM visualized individual macrophages carrying DNMT3A CHIP mutations in human atherosclerotic plaques. Studying monocyte-derived macrophages from Stockholm-Tartu Atherosclerosis Reverse Networks Engineering Task (STARNET; n=941) by WGS revealed CHIP mutations in 14.2% (mean age 67.1 years). RNAseq of these macrophages revealed that expression patterns in CHIP mutation carriers differed substantially from those of non-carriers. Moreover, patterns were different depending on the underlying mutations, e.g. those carrying TET2 mutations predominantly displayed upregulated inflammatory signaling whereas ASXL1 mutations showed stronger effects on metabolic pathways.
Conclusions: Deep-DNA-sequencing reveals a high prevalence of CHIP mutations in whole blood of CAD patients. CHIP-affected leukocytes invade plaques in human coronary arteries. RNAseq data obtained from macrophages of CHIP-affected patients suggest that pro-atherosclerotic signaling differs depending on the underlying mutations. Further studies are necessary to understand whether specific pathways affected by CHIP mutations may be targeted for personalized treatment.
Ilina, Elena ; Armento, Angela ; Sanchez, Leticia ; Reichlmeir, Marina ; Braun, Yannick ; Penski, Cornelia ; Capper, David ; Sahm, Felix ; Jennewein, Lukas ; Harter, Patrick Nikolaus ; Zukunft, Sven ; Fleming, Ingrid ; Schulte, Dorothea ; Guerroué, Francois ; Behrends, Christian ; Ronellenfitsch, Michael Wilfried ; Naumann, Ulrike ; Mittelbronn, Michel Guy André
Carboxypeptidase E (CPE) has recently been described as a multifunctional protein that regulates proliferation, migration and survival in several tumor entities. In glioblastoma (GBM), the most malignant primary brain tumor, secreted CPE (sCPE) was shown to modulate tumor cell migration. In our current study, we aimed at clarifying the underlying molecular mechanisms regulating anti-migratory as well as novel metabolic effects of sCPE in GBM. Here we show that sCPE activates mTORC1 signaling in glioma cells detectable by phosphorylation of its downstream target RPS6. Additionally, sCPE diminishes glioma cell migration associated with a negative regulation of Rac1 signaling via RPS6, since both inhibition of mTOR and stimulation of Rac1 results in a reversed effect of sCPE on migration. Knockdown of CPE leads to a decrease of active RPS6 associated with increased GBM cell motility. Apart from this, we show that sCPE enhances glucose flux into the tricarboxylic acid cycle at the expense of lactate production, thereby decreasing aerobic glycolysis, which might as well contribute to a less invasive behavior of tumor cells. Our data contributes to a better understanding of the complexity of GBM cell migration and sheds new light on how tumor cell invasion and metabolic plasticity are interconnected.
Scholz, Alexander ; Harter, Patrick Nikolaus ; Cremer, Sebastian ; Yalcin, Burak H. ; Gurnik, Stefanie ; Yamaji, Maiko ; Di Tacchio, Mariangela ; Sommer, Kathleen ; Baumgarten, Peter ; Bähr, Roy Oliver ; Steinbach, Joachim Peter ; Trojan, Jörg ; Glas, Martin ; Herrlinger, Ulrich ; Krex, Dietmar ; Meinhardt, Matthias ; Weyerbrock, Astrid ; Timmer, Marco ; Goldbrunner, Roland ; Deckert, Martina ; Braun, Christian ; Schittenhelm, Jens ; Früh, Jochen ; Ullrich, Evelyn ; Mittelbronn, Michel Guy André ; Plate, Karl ; Reiss, Yvonne
Glioblastoma multiforme (GBM) is treated by surgical resection followed by radiochemotherapy. Bevacizumab is commonly deployed for anti‐angiogenic therapy of recurrent GBM; however, innate immune cells have been identified as instigators of resistance to bevacizumab treatment. We identified angiopoietin‐2 (Ang‐2) as a potential target in both naive and bevacizumab‐treated glioblastoma. Ang‐2 expression was absent in normal human brain endothelium, while the highest Ang‐2 levels were observed in bevacizumab‐treated GBM. In a murine GBM model, VEGF blockade resulted in endothelial upregulation of Ang‐2, whereas the combined inhibition of VEGF and Ang‐2 leads to extended survival, decreased vascular permeability, depletion of tumor‐associated macrophages, improved pericyte coverage, and increased numbers of intratumoral T lymphocytes. CD206+ (M2‐like) macrophages were identified as potential novel targets following anti‐angiogenic therapy. Our findings imply a novel role for endothelial cells in therapy resistance and identify endothelial cell/myeloid cell crosstalk mediated by Ang‐2 as a potential resistance mechanism. Therefore, combining VEGF blockade with inhibition of Ang‐2 may potentially overcome resistance to bevacizumab therapy.
Linking epigenetic signature and metabolic phenotype in IDH mutant and IDH wildtype diffuse glioma
(2020)
Braun, Yannick ; Filipski, Katharina Johanna ; Bernatz, Simon ; Baumgarten, Peter ; Roller, Bastian ; Zinke, Jenny ; Zeiner, Pia Susan ; Ilina, Elena ; Senft, Christian ; Ronellenfitsch, Michael Wilfried ; Plate, Karl ; Bähr, Roy Oliver ; Hattingen, Elke ; Steinbach, Joachim Peter ; Mittelbronn, Michel Guy André ; Harter, Patrick Nikolaus
Aims: Changes in metabolism are known to contribute to tumour phenotypes. If and how metabolic alterations in brain tumours contribute to patient outcome is still poorly understood. Epigenetics impact metabolism and mitochondrial function. The aim of this study is a characterisation of metabolic features in molecular subgroups of isocitrate dehydrogenase mutant (IDHmut) and isocitrate dehydrogenase wildtype (IDHwt) gliomas. Methods: We employed DNA methylation pattern analyses with a special focus on metabolic genes, large-scale metabolism panel immunohistochemistry (IHC), qPCR-based determination of mitochondrial DNA copy number and immune cell content using IHC and deconvolution of DNA methylation data. We analysed molecularly characterised gliomas (n = 57) for in depth DNA methylation, a cohort of primary and recurrent gliomas (n = 22) for mitochondrial copy number and validated these results in a large glioma cohort (n = 293). Finally, we investigated the potential of metabolic markers in Bevacizumab (Bev)-treated gliomas (n = 29). Results: DNA methylation patterns of metabolic genes successfully distinguished the molecular subtypes of IDHmut and IDHwt gliomas. Promoter methylation of lactate dehydrogenase A negatively correlated with protein expression and was associated with IDHmut gliomas. Mitochondrial DNA copy number was increased in IDHmut tumours and did not change in recurrent tumours. Hierarchical clustering based on metabolism panel IHC revealed distinct subclasses of IDHmut and IDHwt gliomas with an impact on patient outcome. Further quantification of these markers allowed for the prediction of survival under anti-angiogenic therapy. Conclusion: A mitochondrial signature was associated with increased survival in all analyses, which could indicate tumour subgroups with specific metabolic vulnerabilities.
Obermeier, Martin ; Berg, Thomas ; Sichtig, Nadine ; Braun, Patrick ; Däumer, Martin P. ; Walter, Hauke ; Noah, Christian ; Wolf, Eva ; Müller, Harm ; Stürmer, Martin ; Thielen, Alexander ; Kaiser, Rolf
Poster presentation: Background Maraviroc is a new drug used to treat HIV infection from the new class of drugs called CCR5 entry inhibitors. As the active principle of these drugs is to block the CCR5-receptor on the surface of the target cells, it has to be known if the virus in the patient is using only CCR5 as co-receptor or if there are populations that can also use CXCR4. Therefore, an assay to determine the tropism has to be performed before starting a therapy. Besides phenotypic assays like the TROFILE® assay by Monogram, used in the approval studies, there exist several genotyping systems like geno2pheno-coreceptor, Wetcat (providing five different genotypic tropism schemes) and WebPSSM. ...
Prieto García, Cristian ; Hartmann, Oliver ; Reissland, Michaela ; Braun, Fabian ; Fischer, Thomas ; Walz, Susanne ; Schülein‐Völk, Christina ; Eilers, Ursula ; Ade, Carsten P. ; Calzado, Marco A. ; Orian, Amir ; Maric, Hans-Michael ; Münch, Christian ; Rosenfeldt, Mathias Tillmann ; Eilers, Martin ; Diefenbacher, Markus Elmar
The transcription factor ∆Np63 is a master regulator of epithelial cell identity and essential for the survival of squamous cell carcinoma (SCC) of lung, head and neck, oesophagus, cervix and skin. Here, we report that the deubiquitylase USP28 stabilizes ∆Np63 and maintains elevated ∆NP63 levels in SCC by counteracting its proteasome‐mediated degradation. Impaired USP28 activity, either genetically or pharmacologically, abrogates the transcriptional identity and suppresses growth and survival of human SCC cells. CRISPR/Cas9‐engineered in vivo mouse models establish that endogenous USP28 is strictly required for both induction and maintenance of lung SCC. Our data strongly suggest that targeting ∆Np63 abundance via inhibition of USP28 is a promising strategy for the treatment of SCC tumours.
Lower leg fractures in children and adolescents - comparison of conservative vs. ECMES treatment
(2021)
Weber, Birte ; Kalbitz, Miriam ; Baur, Meike ; Braun, Christian Karl ; Zwingmann, Jörn ; Preßmar, Jochen
Background: Lower leg fractures are one of the most common fractures in pediatric age. In general, treatment of lower leg fractures is predominantly non-operative, requiring clinical and radiological controls. Nevertheless, it can be observed that in recent years tibial shaft fractures have increasingly been treated surgically. The aim of the present study is to investigate treatment strategies in the context of different fracture types of the lower leg Methods: In this retrospective chart review, we analyzed 168 children with a diaphyseal fracture of the lower leg admitted to a trauma center between 2005 and 2017. The fractures were classified according to the AO Pediatric Comprehensive Classification of Long Bone Fractures (AO-PCCF). Results: The frequency of fractures based on the AO-PCCF classification was as follows: Simple oblique fracture of the tibia (43.5%, n = 73), hereof 32 toddler's fractures, multifragmentary oblique fracture of the tibia in 14.3% (n = 24) and simple oblique fracture of both, tibia and fibula in 18 patients (10.7%). Most pediatric fractures were treated conservatively by cast (n = 125). Thirty-seven patients received an ECMES, whereas 3 patients were treated with an external fixator and also 3 fractures were stabilized by plate osteosynthesis. Conservatively treated patients were significantly younger (mean age 6.0) compared to patients treated with ECMES (mean age 10.2) or plate osteosynthesis (PO)/external fixator (EF) (mean age 11.3), even if toddler's fractures (mean age 2.0) are excluded (mean age 7.4). There was no difference in time to full weight-bearing, hospitalization of patients treated with ECMES compared to conservative therapy although ECMES-treated fractures show more instability. The consolidation time was significantly higher in ECMES treated patients compared to conservative therapy. Conclusion: Pediatric patients (≤4 years) with lower leg fractures most often showed simple oblique fractures of the tibia, half of them toddler's fractures, which were treated predominantly by conservative therapy. All in all, the consolidation time was longer in intramedullary nailing (ECMES) than in conservative therapy. Nevertheless, time to full weight bearing and duration of cast was the same in both groups, even though ECMES treated fractures show more instability.