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Leblond, Claire S. ; Heinrich, Jutta ; Delorme, Richard ; Proepper, Christian ; Betancur, Catalina ; Huguet, Guillaume ; Konyukh, Marina ; Chaste, Pauline ; Ey, Elodie ; Rastam, Maria ; Anckarsäter, Henrik ; Nygren, Gudrun ; Gillberg, I. Carina ; Melke, Jonas ; Toro, Roberto ; Regnault, Beatrice ; Fauchereau, Fabien ; Mercati, Oriane ; Lemière, Nathalie ; Skuse, David ; Poot, Martin ; Holt, Richard ; Monaco, Anthony P. ; Järvelä, Irma ; Kantojärvi, Katri ; Vanhala, Raija ; Curran, Sarah ; Collier, David A. ; Bolton, Patrick F. ; Geburtig-Chiocchetti, Andreas ; Klauck, Sabine M. ; Poustka, Fritz ; Freitag, Christine M. ; Waltes, Regina ; Kopp, Marnie ; Duketis, Eftichia ; Bacchelli, Elena ; Minopoli, Fiorella ; Ruta, Liliana ; Battaglia, Agatino ; Mazzone, Luigi ; Maestrini, Elena ; Sequeira, Ana F. ; Oliveira, Barbara ; Vicente, Astrid M. ; Oliveira, Guiomar ; Pinto, Dalila ; Scherer, Stephen W. ; Zelenika, Diana ; Delepine, Marc ; Lathrop, Mark ; Bonneau, Dominique ; Guinchat, Vincent ; Devillard, Françoise ; Assouline, Brigitte ; Mouren, Marie-Christine ; Leboyer, Marion ; Gillberg, Christopher ; Böckers, Tobias M. ; Bourgeron, Thomas
Autism spectrum disorders (ASD) are a heterogeneous group of neurodevelopmental disorders with a complex inheritance pattern. While many rare variants in synaptic proteins have been identified in patients with ASD, little is known about their effects at the synapse and their interactions with other genetic variations. Here, following the discovery of two de novo SHANK2 deletions by the Autism Genome Project, we identified a novel 421 kb de novo SHANK2 deletion in a patient with autism. We then sequenced SHANK2 in 455 patients with ASD and 431 controls and integrated these results with those reported by Berkel et al. 2010 (n = 396 patients and n = 659 controls). We observed a significant enrichment of variants affecting conserved amino acids in 29 of 851 (3.4%) patients and in 16 of 1,090 (1.5%) controls (P = 0.004, OR = 2.37, 95% CI = 1.23-4.70). In neuronal cell cultures, the variants identified in patients were associated with a reduced synaptic density at dendrites compared to the variants only detected in controls (P = 0.0013). Interestingly, the three patients with de novo SHANK2 deletions also carried inherited CNVs at 15q11-q13 previously associated with neuropsychiatric disorders. In two cases, the nicotinic receptor CHRNA7 was duplicated and in one case the synaptic translation repressor CYFIP1 was deleted. These results strengthen the role of synaptic gene dysfunction in ASD but also highlight the presence of putative modifier genes, which is in keeping with the "multiple hit model" for ASD. A better knowledge of these genetic interactions will be necessary to understand the complex inheritance pattern of ASD.
A toolbox for the generation of chemical probes for Baculovirus IAP Repeat containing proteins
(2022)
Schwalm, Martin P. ; Berger, Lena Marie ; Meuter, Maximilian N. ; Vasta, James D. ; Corona, Cesear R. ; Röhm, Sandra ; Berger, Benedict-Tilman ; Farges, Frederic ; Beinert, Sebastian M. ; Preuß, Franziska Friederike ; Morasch, Viktoria ; Rogov, Vladimir V. ; Mathea, Sebastian ; Saxena, Krishna ; Robers, Matthew B. ; Müller, Susanne ; Knapp, Stefan
E3 ligases constitute a large and diverse family of proteins that play a central role in regulating protein homeostasis by recruiting substrate proteins via recruitment domains to the proteasomal degradation machinery. Small molecules can either inhibit, modulate or hijack E3 function. The latter class of small molecules led to the development of selective protein degraders, such as PROTACs (PROteolysis TArgeting Chimeras), that recruit protein targets to the ubiquitin system leading to a new class of pharmacologically active drugs and to new therapeutic options. Recent efforts have focused on the E3 family of Baculovirus IAP Repeat (BIR) domains that comprise a structurally conserved but diverse 70 amino acid long protein interaction domain. In the human proteome, 16 BIR domains have been identified, among them promising drug targets such as the Inhibitors of Apoptosis (IAP) family, that typically contain three BIR domains (BIR1, BIR2, and BIR3). To date, this target area lacks assay tools that would allow comprehensive evaluation of inhibitor selectivity. As a consequence, the selectivity of current BIR domain targeting inhibitors is unknown. To this end, we developed assays that allow determination of inhibitor selectivity in vitro as well as in cellulo. Using this toolbox, we have characterized available BIR domain inhibitors. The characterized chemical starting points and selectivity data will be the basis for the generation of new chemical probes for IAP proteins with well-characterized mode of action and provide the basis for future drug discovery efforts and the development of PROTACs and molecular glues.
Schrod, Jann Erik ; Thomson, Erik S. ; Weber, Daniel ; Koßmann, Jens ; Pöhlker, Christopher ; Saturno, Jorge ; Ditas, Florian ; Artaxo, Paulo ; Clouard, Valérie ; Saurel, Jean-Marie ; Ebert, Martin ; Curtius, Joachim ; Bingemer, Heinz
Ice particle activation and evolution have important atmospheric implications for cloud formation, initiation of precipitation and radiative interactions. The initial formation of atmospheric ice by heterogeneous ice nucleation requires the presence of a nucleating seed, an ice-nucleating particle (INP), to facilitate its first emergence. Unfortunately, only a few long-term measurements of INPs exist, and as a result, knowledge about geographic and seasonal variations of INP concentrations is sparse. Here we present data from nearly 2 years of INP measurements from four stations in different regions of the world: the Amazon (Brazil), the Caribbean (Martinique), central Europe (Germany) and the Arctic (Svalbard). The sites feature diverse geographical climates and ecosystems that are associated with dissimilar transport patterns, aerosol characteristics and levels of anthropogenic impact (ranging from near pristine to mostly rural). Interestingly, observed INP concentrations, which represent measurements in the deposition and condensation freezing modes, do not differ greatly from site to site but usually fall well within the same order of magnitude. Moreover, short-term variability overwhelms all long-term trends and/or seasonality in the INP concentration at all locations. An analysis of the frequency distributions of INP concentrations suggests that INPs tend to be well mixed and reflective of large-scale air mass movements. No universal physical or chemical parameter could be identified to be a causal link driving INP climatology, highlighting the complex nature of the ice nucleation process. Amazonian INP concentrations were mostly unaffected by the biomass burning season, even though aerosol concentrations increase by a factor of 10 from the wet to dry season. Caribbean INPs were positively correlated to parameters related to transported mineral dust, which is known to increase during the Northern Hemisphere summer. A wind sector analysis revealed the absence of an anthropogenic impact on average INP concentrations at the site in central Europe. Likewise, no Arctic haze influence was observed on INPs at the Arctic site, where low concentrations were generally measured. We consider the collected data to be a unique resource for the community that illustrates some of the challenges and knowledge gaps of the field in general, while specifically highlighting the need for more long-term observations of INPs worldwide.
Yalachkov, Yavor Vasilev ; Dabanli, Dilara ; Wenger-Alakmeh, Katharina ; Forster, Marie-Thérèse ; Steinbach, Joachim Peter ; Voss, Martin
Introduction: The concurrent presence of both central nervous system (CNS) tumors and multiple sclerosis (MS) poses various diagnostic and therapeutic pitfalls and makes the clinical management of such patients challenging.
Methods: In this retrospective, single-center cohort study, we searched our clinical databases (2006–2019) for patients with concurrent CNS tumors and MS and described their disease courses. Age at diagnosis of the respective disease and probabilities for MS disease activity events (DAEs) with vs. without prior tumor-specific therapy were tested pairwise using t-test for dependent samples and exact binomial test.
Results: N = 16 patients with concurrent CNS tumors and MS were identified. MS diagnosis preceded the CNS oncological diagnosis by an average of 9 years (p = 0.004). More DAEs occurred in patients without prior chemotherapy (83.3%) than in patients with prior chemotherapy (16.7%; p = 0.008). This effect did not reach significance for patients with prior radiation therapy/radiosurgery (66.7% vs. 33.3%, p = 0.238). The average interval between DAEs and the last documented lymphopenia was 32.25 weeks.
Conclusions: This study describes the clinical and demographic features of patients with concurrent CNS tumors and MS and suggests several practical approaches to their clinical management. Our findings suggest that adding a disease-modifying MS therapy to the regimen of patients treated with chemotherapy is necessary only if the patient suffers from a highly active, aggressive course of MS. In view of the lack of prospective trials, individual risk assessments should remain the foundation of the decision on MS treatment in concurrent CNS tumor diseases.
Forster, Marie-Thérèse ; Behrens, Marion ; Lortz, Irina ; Conradi, Nadine Caroline ; Senft, Christian ; Voß, Martin ; Rauch, Maximilian ; Seifert, Volker
Due to anticipated postoperative neuropsychological sequelae, patients with gliomas infiltrating the corpus callosum rarely undergo tumor resection and mostly present in a poor neurological state. We aimed at investigating the benefit of glioma resection in the corpus callosum, hypothesizing neuropsychological deficits were mainly caused by tumor presence. Between 01/2017 and 1/2020, 21 patients who underwent glioma resection in the corpus callosum were prospectively enrolled into this study. Neuropsychological function was assessed preoperatively, before discharge and after 6 months. Gross total tumor resection was possible in 15 patients, and in 6 patients subtotal tumor resection with a tumor reduction of 97.7% could be achieved. During a median observation time of 12.6 months 9 patients died from glioblastoma after a median of 17 months. Preoperatively, all cognitive domains were affected in up to two thirds of patients, who presented a median KPS of 100% (range 60–100%). After surgery, the proportion of impaired patients increased in all neurocognitive domains. Most interestingly, after 6 months, significantly fewer patients showed impairments in attention, executive functioning, memory and depression, which are domains considered crucial for everyday functionality. Thus, the results of our study strongly support our hypothesis that in patients with gliomas infiltrating the corpus callosum the benefit of tumor resection might outweigh morbidity.
Schmidt, Florian ; Marx, Alexander ; Hebel, Marie ; Wegner, Martin ; Baumgarten, Nina ; Kaulich, Manuel ; Göke, Jonathan ; Vreeken, Jilles ; Schulz, Marcel Holger
Understanding the complexity of transcriptional regulation is a major goal of computational biology. Because experimental linkage of regulatory sites to genes is challenging, computational methods considering epigenomics data have been proposed to create tissue-specific regulatory maps. However, we showed that these approaches are not well suited to account for the variations of the regulatory landscape between cell-types. To overcome these drawbacks, we developed a new method called STITCHIT, that identifies and links putative regulatory sites to genes. Within STITCHIT, we consider the chromatin accessibility signal of all samples jointly to identify regions exhibiting a signal variation related to the expression of a distinct gene. STITCHIT outperforms previous approaches in various validation experiments and was used with a genome-wide CRISPR-Cas9 screen to prioritize novel doxorubicin-resistance genes and their associated non-coding regulatory regions. We believe that our work paves the way for a more refined understanding of transcriptional regulation at the gene-level.
Willems, Laurent Maximilian ; Bertsche, Astrid ; Bösebeck, Frank ; Hornemann, Frauke ; Immisch, Ilka ; Klein, Karl Martin ; Knake, Susanne ; Kunz, Rhina ; Kurlemann, Gerhard ; Langenbruch, Lisa Marie ; Möddel, Gabriel ; Müller-Schlüter, Karen ; Podewils, Felix von ; Reif, Philipp Sebastian ; Steinhoff, Bernhard J. ; Steinig, Isabel ; Rosenow, Felix ; Schubert-Bast, Susanne ; Strzelczyk, Adam
Objective: To evaluate the efficacy and tolerability of brivaracetam (BRV) in a severely drug refractory cohort of patients with epileptic encephalopathies (EE).
Method: A multicenter, retrospective cohort study recruiting all patients treated with EE who began treatment with BRV in an enrolling epilepsy center between 2016 and 2017.
Results: Forty-four patients (27 male [61%], mean age 29 years, range 6 to 62) were treated with BRV. The retention rate was 65% at 3 months, 52% at 6 months and 41% at 12 months. A mean retention time of 5 months resulted in a cumulative exposure to BRV of 310 months. Three patients were seizure free during the baseline. At 3 months, 20 (45%, 20/44 as per intention-to-treat analysis considering all patients that started BRV including three who were seizure free during baseline) were either seizure free (n = 4; 9%, three of them already seizure-free at baseline) or reported at least 25% (n = 4; 9%) or 50% (n = 12; 27%) reduction in seizures. An increase in seizure frequency was reported in two (5%) patients, while there was no change in the seizure frequency of the other patients. A 50% long-term responder rate was apparent in 19 patients (43%), with two (5%) free from seizures for more than six months and in nine patients (20%, with one [2 %] free from seizures) for more than 12 months. Treatment-emergent adverse events were predominantly of psychobehavioural nature and were observed in 16%.
Significance: In this retrospective analysis the rate of patients with a 50% seizure reduction under BRV proofed to be similar to those seen in regulatory trials for focal epilepsies. BRV appears to be safe and relatively well tolerated in EE and might be considered in patients with psychobehavioral adverse events while on levetiracetam.
Queck, Alexander David Roger ; Bode, Hannah ; Uschner, Frank Erhard ; Brol, Maximilian ; Graf, Christiana ; Schulz, Martin ; Jansen, Christian ; Praktiknjo, Michael ; Schierwagen, Robert ; Klein, Sabine ; Trautwein, Christian ; Wasmuth, Hermann E. ; Berres, Marie-Luise ; Trebicka, Jonel ; Lehmann, Jennifer
Background and Aims: Monocyte chemotactic protein-1 (MCP-1) is a potent chemoattractant for monocytes. It is involved in pathogenesis of several inflammatory diseases. Hepatic MCP-1 is a readout of macrophage activation. While inflammation is a major driver of liver disease progression, the origin and role of circulating MCP-1 as a biomarker remains unclear.
Methods: Hepatic CC-chemokine ligand 2 (CCL2) expression and F4/80 staining for Kupffer cells were measured and correlated in a mouse model of chronic liver disease (inhalative CCl4 for 7 weeks). Next, hepatic RNA levels of CCL2 were measured in explanted livers of 39 patients after transplantation and correlated with severity of disease. Changes in MCP-1 were further evaluated in a rat model of experimental cirrhosis and acute-on-chronic liver failure (ACLF). Finally, we analyzed portal and hepatic vein levels of MCP-1 in patients receiving transjugular intrahepatic portosystemic shunt insertion for complications of portal hypertension.
Results: In this mouse model of fibrotic hepatitis, hepatic expression of CCL2 (P = 0.009) and the amount of F4/80 positive cells in the liver (P < 0.001) significantly increased after induction of hepatitis by CCl4 compared to control animals. Moreover, strong correlation of hepatic CCL2 expression and F4/80 positive cells were seen (P = 0.023). Furthermore, in human liver explants, hepatic transcription levels of CCL2 correlated with the MELD score of the patients, and thus disease severity (P = 0.007). The experimental model of ACLF in rats revealed significantly higher levels of MCP-1 plasma (P = 0.028) and correlation of hepatic CCL2 expression (R = 0.69, P = 0.003). Particularly, plasma MCP-1 levels did not correlate with peripheral blood monocyte CCL2 expression. Finally, higher levels of MCP-1 were observed in the hepatic compared to the portal vein (P = 0.01) in patients receiving TIPS. Similarly, a positive correlation of MCP-1 with Child-Pugh score was observed (P = 0.018). Further, in the presence of ACLF, portal and hepatic vein levels of MCP-1 were significantly higher compared to patients without ACLF (both P = 0.039).
Conclusion: Circulating levels of MCP-1 mainly derive from the injured liver and are associated with severity of liver disease. Therefore, liver macrophages contribute significantly to disease progression. Circulating MCP-1 may reflect the extent of hepatic macrophage activation.
Assfalg, Volker ; Selig, Katharina ; Tolksdorf, Johanna ; Meel, Marieke van ; Vries, Erwin de ; Ramsoebhag, Anne‐Marie ; Rahmel, Axel ; Renders, Lutz ; Novotny, Alexander ; Matevossian, Edouard ; Schneeberger, Stefan ; Rosenkranz, Alexander ; Berlakovich, Gabriela ; Ysebaert, Dirk ; Knops, Noël ; Kuypers, Dirk ; Weekers, Laurent ; Muehlfeld, Anja ; Rump, Lars‐Christian ; Hauser, Ingeborg A. ; Pisarski, Przemyslaw ; Weimer, Rolf ; Fornara, Paolo ; Fischer, Lutz ; Kliem, Volker ; Sester, Urban ; Stippel, Dirk ; Arns, Wolfgang ; Hau, Hans‐Michael ; Nitschke, Martin ; Hoyer, Joachim ; Thorban, Stefan ; Weinmann‐Menke, Julia ; Heller, Katharina ; Banas, Bernhard ; Schwenger, Vedat ; Nadalin, Silvio ; Lopau, Kai ; Hüser, Norbert ; Heemann, Uwe
In Eurotransplant kidney allocation system (ETKAS), candidates can be considered unlimitedly for repeated re‐transplantation. Data on outcome and benefit are indeterminate. We performed a retrospective 15‐year patient and graft outcome data analysis from 1464 recipients of a third or fourth or higher sequential deceased donor renal transplantation (DDRT) from 42 transplant centers. Repeated re‐DDRT recipients were younger (mean 43.0 vs. 50.2 years) compared to first DDRT recipients. They received grafts with more favorable HLA matches (89.0% vs. 84.5%) but thereby no statistically significant improvement of patient and graft outcome was found as comparatively demonstrated in 1st DDRT. In the multivariate modeling accounting for confounding factors, mortality and graft loss after 3rd and ≥4th DDRT (P < 0.001 each) and death with functioning graft (DwFG) after 3rd DDRT (P = 0.001) were higher as compared to 1st DDRT. The incidence of primary nonfunction (PNF) was also significantly higher in re‐DDRT (12.7%) than in 1st DDRT (7.1%; P < 0.001). Facing organ shortage, increasing waiting time, and considerable mortality on dialysis, we question the current policy of repeated re‐DDRT. The data from this survey propose better HLA matching in first DDRT and second DDRT and careful selection of candidates, especially for ≥4th DDRT.