Characterization of proteomic, metabolomic and functional responses in endothelial cells after long-term exposure to diesel soot surrogates

Zielsetzung der Arbeit war, molekulare bzw. zelluläre Effekte an endothelialen Zellen nach Langzeitexposition gegen die Dieselrußsurrogate Carbon Black und Benzo[a]pyren-beladenes Carbon Black im Niedrigdosisbereich zu ermitteln. Dieselruß ist in Abhängigkeit vom Entstehungsort mit einer Vielzahl von Substanzen wie Übergangsmetallen (z. B. Eisen, Chrom), Salzen und organischen Verbindungen (z. B. PAH) belastet, die selbst zelluläre Effekte auslösen können. Daher sollten in dieser Arbeit die Zellen gegen die Diesel¬ruß-surrogate Carbon Black und B[a]P-beladenes Carbon Black exponiert werden. Benzo[a]pyren (Leitsubstanz der PAH) sollte stellvertretend für die Gruppe der polyzyklischen aromatischen Kohlenwasserstoffe die Kontamination von Dieselruß mit dieser Stoffgruppe simulieren. Diskutierte Mechanismen zur Entwicklung von Feinstaub-vermittelten kardiovaskulären Erkrankungen beruhen auf Experimenten, bei denen meist Ultrafeinstaub¬partikel in hohen Dosen über einen kurzen Zeitraum eingesetzt wurden. Die Exposition des Menschen erfolgt im Gegensatz zu den gängigen experimentellen Studien über einen längeren Zeitraum gegen geringe Dosen. Aus diesem Grunde wurden in der vorliegenden Arbeit ein Expositionszeitraum von 14 Tagen sowie eine Exposition im Niedrigdosisbereich gewählt. Die Exposition der Endothelzellen gegen eine niedrigere bzw. physiologischere Konzentration von Dieselru߬surrogaten steigerte die Proliferation und veränderte den Energiehaushalt, jedoch nicht die Generierung von ROS. Diese Antworten ließen sich anhand der Ergebnisse der proteomischen Analyse exponierter Zellen zu einem großen Teil auf eine verringerte Aktivität des Transkriptions¬faktors PPARγ zurückführen. Die Hypothese, dass PPARγ ein wichtiger Regulator der beobachteten zellulären Reaktionen ist, wurde durch die gas¬chromato¬graphische Analyse der zellulären Metaboliten erhärtet. So wurden die in der Literatur beschriebenen Veränderungen im Energiestoffwechsel infolge einer verminderten PPARγ-Aktivität in der vorliegenden Arbeit anhand des Nachweises einer erhöhten intrazellulären Konzentration an freien Fettsäuren und eines durch die gesteigerte Proliferation erhöhten Energiebedarfs bestätigt. Da zu erwarten war, dass die bereits identifizierten Proteine nicht die Gesamtheit der beeinflussten Signalwege repräsentieren, wurde zusätzlich das Phosphoproteom untersucht. Die Analyse phosphorylierter Proteine ist jedoch eine schwierige Aufgabe in Anbetracht ihres geringen Vorkommens. Die Anreicherung der Phosphoproteine schien dabei als ein vielversprechender Ansatz, verfügbare Techniken setzten jedoch hoch¬technisierte Geräte voraus. Daher wurde ein Verfahren zur einfachen Anreicherung mittels Lanthanionen (La3+) entwickelt, durch dessen Anwendung Phosphoproteine und Phospho¬glykoproteine selektiv aus komplexen Proteingemischen isoliert werden konnten. Die Analyse des Phosphoproteoms endothelialer Zellen nach Exposition ergab Änderungen der Phosphorylierung von Proteinen, die neben katalytischen Prozessen auch die Struktur der Zellen beeinflussten. Die Modifikationen der zytoskelettalen Proteine wiesen auf ein erhöhtes Angio¬genese¬potential hin, was sich in funktionellen Tests (Migration, Invasion, Barriere-funktion) bestätigte. Der Anteil migrierender Zellen sowie ihre Invasionsbereitschaft erhöhten sich nach Exposition gegen die Dieselrußsurrogate. Darüber hinaus wiesen die exponierten Zellen eine verringerte Permeabilität gegenüber geladenen und ungeladenen Substanzen (Dextrane) auf. Die funktionellen Veränderungen spiegelten sich auch auf morphologischer Ebene in der Ausbildung von Filopodien und Stressfasern wider. Als möglicher Auslöser dieser zellulären Veränderungen kommt der erhöhte Gehalt an intrazellulärem Calcium infrage. Calcium stellt einen wichtigen zellulären Signaltransduktor dar, der an vielen Prozessen, darunter der Regulation der endothelialen Stickstoffmonoxid-Produktion, beteiligt ist. Die endotheliale NO-Produktion ist ein wichtiger Modulator der Angiogenese, die viele der beobachteten Effekte beeinflussen kann. Die hier vorgestellten Ergebnisse beschreiben detaillierte Veränderungen des Endothels als Antwort auf eine Exposition gegen Dieselrußsurrogate. Ob und ggf. welche/s Krankheits-bild/er diese Veränderungen hervorrufen können, ist zum jetzigen Zeitpunkt noch unklar.Nanotechnology is a rapidly growing industry of global economic importance, exploiting the novel characteristics of materials manufactured at the nanoscale. The properties of engineered nanoparticles (ENPs) that make them useful in a wide range of industrial applications, however, have led to concerns regarding their potential impact on human and environmental health. Nanometer sized particles are created in countless physical processes ranging from natural calamities such as erosion and combustion to many industrial and domestic endeavours, such as cooking, material fabrication, jet engines and in intentionally engineered nanoscale components of consumer products and advanced technologies. The impact of nanoparticles interactions with the body is dependent on their size, chemical composition, surface structure, solubility and shape. Due to small size and hence higher specific surface area of the nanoparticles, these particles can easily bind with toxic pollutants such as transition metals and organic pollutants (i.e. polycyclic aromatic hydrocarbons (PAH)), which when inhaled can cause a number of diseases. Inhaled nanoparticles have the ability to translocate in the body. Once inside, these particles can travel freely in the blood throughout the body and reach the organs like liver, heart and brain. Epidemiologic studies have indicated that environmental exposure to airborne particulate matter (primarily diesel exhaust) may promote cardiovascular diseases; however, it is not clear whether these effects are due to the exposure to these environment particles or because of its co-contaminants (i.e. iron, chromium, salts and PAHs). The precise ways of action are still unknown and the suggested mechanisms often lack the convincing evidence because they have been derived from experiments in which nanoparticles have been applied in high doses. In the present study the human endothelial cell line EA.hy926 was exposed to pure carbon black and benzo[a]pyrene loaded carbon black to mimic diesel exhaust, to determine the effects of these nanoparticles on the cardiovascular system. To achieve this, a proteomic and metabolomic approach was carried out with particular emphasis on an extended exposure period of 14 days and a low nanoparticle concentration of 100 ng/mL. Evaluation of the obtained proteomic data combined with transcription factor analysis revealed that the activity of the peroxisome proliferator-activated receptor gamma (PPARγ) was significantly reduced in the exposed cells. This result was in agreement with the observed profile of energy and fatty acid metabolites determined by gas chromatographic mass spectrometric analysis. Given that the previously identified proteins did not represent the entirety of the affected signaling pathways, therefore, possible posttranslational modifications of the exposed endothelial cells was examined. Phosphorylation is one of the most universal and dominant mechanism involved in the regulation of cellular metabolism. Due to the low abundance of phosphorylated proteins there is a great interest in developing methods for their enrichment. Using lanthanum ions, a simple technique for phosphoprotein enrichment was established. The method not only allowed the selective enrichment of phospho- and phosphoglycoproteins from complex protein mixtures, but also was compatible with analytical techniques. The analysis of the phosphoproteome of endothelial cells after exposure revealed changes in the phosphorylation of proteins involved in cell structure maintenance. These changes in the phosphorylation referred thereby to modulations in the angiogenic system which could be shown by an increased cell invasion and migration, and structural changes of the actin cytoskeleton. The trigger of these cellular responses is possible calcium which was present in elevated levels in exposed cell and may influence the endothelial NO production

Overcoming a Communication Barrier on the Way Towards a Global Sensor Network

In a global sensor network different sensor platforms will be deployed. A grave obstacle on the way of building sensor networks out of different sensor nodes are incompatible implementations of network protocol stacks used with different sensor node platforms. We describe our efforts to overcome this obstacle in a heterogeneous sensor network consisting out of MICAz Motes and Sun SPOTs, both using an IEEE 802.15.4 radio chip. We explain the major differences in the respective network stacks and our approach to bridge them. A network stack that bridges the gap between different platforms allows for more flexible and robust networks

A new perspective on calmodulin-regulated calcium and ROS homeostasis upon carbon black nanoparticle exposure

Toxicological studies propose that exposure to carbon black nanoparticles induces organ injuries and inflammatory responses. Besides, current understanding of the molecular mechanisms implies that carbon black nanoparticles (CBNP) exposure induces the production of reactive oxygen species (ROS) causing inflammation, mitochondrial dysfunction or disturbance in calcium homeostasis. However, the precise mechanisms whereby CBNP exert these effects in the lung are still not fully understood. To gain insight into the possible mechanism of CBNP exerted toxicity, human alveolar epithelial cells (A549) were exposed to different concentrations of CBNP and for different timepoints. The reaction of the cells was monitored by the systematic use of cell-based measurements of calcium and ROS, in the presence and absence of calcium (Ca2+) pump inhibitors/chelators and antioxidants. Followed by an in-depth PCR analysis of 84 oxidative stress-related genes. The measurements revealed, as compared to the control, that exposure to CBNP nanoparticles leads to the generation of high ROS levels, as well as a disturbance in calcium homeostasis, which remained primarily unchanged even after 24 h of exposure. Nevertheless, in presence of antioxidants N-acetylcysteine (NAC) and Trolox, ROS formation was considerably reduced without affecting the intracellular calcium concentration. On the other hand, Ca2+ pump inhibitors/chelators, BAPTA (1,2-bis(o-amino phenoxy)ethane-N, N, N′, N′-tetraacetic acid) and verapamil not only decreased the Ca2+ overload, but also further decreased the ROS formation, indicating its role in CBNP-induced oxidative stress. Further, a PCR array analysis of A549 cells in presence and absence of the calmodulin (CaM) antagonist W7, indicated toward nine altered oxidative stress-related genes which further confirmed our cytotoxicity results. Obtained data suggested that CBNP exposure elevates calcium ion concentration, which further contributes to oxidative stress, via the calcium-binding protein CaM. Its inhibition with W7 leads to downregulation in gene expression of nine oxidative stress-related genes, which otherwise, as compared to control, show increased gene expression. The results of the study thus confirm that exposure of lung epithelial cells to CBNP leads to oxidative stress; however, the oxidative stress itself is a result of a disturbance in both calcium and ROS homeostasis, and should be considered while searching for a new strategy for prevention of CBNP-induced lung toxicity

Towards a Realistic Energy Model for Wireless Sensor Networks

Energy consumption is one of the most critical protocol properties in a wireless sensor network. Therefore, a precise energy model is required for the evaluation of protocols. In addition to evaluation, an energy model can also be used for on-line energy accounting. After processing user queries, nodes aware of the energy model can send an energy bill towards the network\u27s sink. This allows the user to adjust future queries to be more energy efficient. The contribution of this paper is twofold: First, a theoretical energy model based on simple finite automata is presented. This model can be used for on-line accounting, simulation and generation of a-priori knowledge. Second, the proposed theoretical model is backed up by practical measurements using a new measurement device called SNMD that also offers management functions for sensor-net testbeds. Therewith, output of the theoretical model can be compared to and justified by real-world measurements

Assessment of the different skin sensitization potentials of irritants and allergens as single substances and in combination using the KeratinoSens assay

Abstract Background People are exposed to mixtures containing allergens and irritants often causing contact dermatitis. Therefore, regulatory authorities require systematic information on the effects of mixtures on the sensitization threshold. In this study a moderate (cinnamal) and a weak (ethylene glycol dimethacrylate) allergen were combined with irritants covering different mechanisms of action (sodium dodecyl sulfate, salicylic acid, and α‐pinene). For a systematic approach, the single substances were initially tested using the KeratinoSens assay. Thereafter, each allergen was combined with noncytotoxic concentrations of the irritants. Method The KeratinoSens assay was applied for the single substances according to OECD (Organisation for Economic Co‐operation and Development) Test Guideline 442D. Based on these results, three noncytotoxic concentrations of the irritants were selected and applied simultaneously with 12 concentrations of the allergens to the KeratinoSens cells. Sensitization threshold and cytotoxicity were measured and compared with the individual testing. Results The combinations of allergens and irritants differed from the effects of the single substances and lowered the sensitization threshold. The quantitative approach allowed a clear description of the changes which varied by factors between 1.1 and 10.3. Conclusions Overall, the allergen was the prominent compound in the mixture and its nature appeared to determine the degree of the response

How can we justify grouping of nanoforms for hazard assessment? Concepts and tools to quantify similarity

The risk of each nanoform (NF) of the same substance cannot be assumed to be the same, as they may vary in their physicochemical characteristics, exposure and hazard. However, neither can we justify a need for more animal testing and resources to test every NF individually. To reduce the need to test all NFs, (regulatory) information requirements may be fulfilled by grouping approaches. For such grouping to be acceptable, it is important to demonstrate similarities in physicochemical properties, toxicokinetic behaviour, and (eco)toxicological behaviour. The GRACIOUS Framework supports the grouping of NFs, by identifying suitable grouping hypotheses that describe the key similarities between different NFs. The Framework then supports the user to gather the evidence required to test these hypotheses and to subsequently assess the similarity of the NFs within the proposed group. The evidence needed to support a hypothesis is gathered by an Integrated Approach to Testing and Assessment (IATA), designed as decision trees constructed of decision nodes. Each decision node asks the questions and provides the methods needed to obtain the most relevant information. This White paper outlines existing and novel methods to assess similarity of the data generated for each decision node, either via a pairwise analysis conducted property-by-property, or by assessing multiple decision nodes simultaneously via a multidimensional analysis. For the pairwise comparison conducted property-by-property we included in this White paper: • A Bayesian model assessment which compares two sets of values using nested sampling. This approach is new in NF grouping. • A Arsinh-Ordered Weighted Average model (Arsinh-OWA) which applies the arsinh transformation to the distance between two NFs, and then rescales the result to the arsinh of a biologically relevant threshold before grouping using OWA based distance. This approach is new in NF grouping. • An x-fold comparison as used in the ECETOC NanoApp. • Euclidean distance, which is a highly established distance metric. The x-fold, Bayesian and Arsinh-OWA distance algorithms performed comparably in the scoring of similarity between NF pairs. The Euclidean distance was also useful, but only with proper data transformation. The x-fold method does not standardize data, and thus produces skewed histograms, but has the advantage that it can be implemented without programming knowhow. A range of multidimensional evaluations, using for example dendrogram clustering approaches, were also investigated. Multidimensional distance metrics were demonstrated to be difficult to use in a regulatory context, but from a scientific perspective were found to offer unexpected insights into the overall similarity of very different materials. In conclusion, for regulatory purposes, a property-by-property evaluation of the data matrix is recommended to substantiate grouping, while the multidimensional approaches are considered to be tools of discovery rather than regulatory methods

Film as architectural theory

Publications on architectural theory have predominantly taken on the form of text-based books, monographs, and articles. With the rise of transdisciplinary and practice-based research in architecture, new opportunities are opening up for other forms of architectural theory, such as film-based mediums, which promise to expand and alter the convention of the written practice of theory. Two possible types of filmic theory are presented here. One follows the method of ethnographic documentary filmmaking inspired by Sarah Pinkfilm-based mediums, which promise to expand and alter thellows the line of art house filmmaking inspired by Kathryn Rameyyn Rameyg inspired by Sarah Pinkfilm-based mediums, which promise to expand ae to expand ad mediums, which promise to expand a convention of the written practice of theory. or constructing knowledge, new discourses on filmic theory can be opened up. It is argued here that film as architectural theory is part of this new discourse, broadening the audience’u engagement with architecture through not only “readership” but also “viewership.

Minimal information for studies of extracellular vesicles 2018 (MISEV2018):a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines

The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points

Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly

Detailed stratified GWAS analysis for severe COVID-19 in four European populations

Given the highly variable clinical phenotype of Coronavirus disease 2019 (COVID-19), a deeper analysis of the host genetic contribution to severe COVID-19 is important to improve our understanding of underlying disease mechanisms. Here, we describe an extended genome-wide association meta-analysis of a well-characterized cohort of 3255 COVID-19 patients with respiratory failure and 12 488 population controls from Italy, Spain, Norway and Germany/Austria, including stratified analyses based on age, sex and disease severity, as well as targeted analyses of chromosome Y haplotypes, the human leukocyte antigen region and the SARS-CoV-2 peptidome. By inversion imputation, we traced a reported association at 17q21.31 to a ~0.9-Mb inversion polymorphism that creates two highly differentiated haplotypes and characterized the potential effects of the inversion in detail. Our data, together with the 5th release of summary statistics from the COVID-19 Host Genetics Initiative including non-Caucasian individuals, also identified a new locus at 19q13.33, including NAPSA, a gene which is expressed primarily in alveolar cells responsible for gas exchange in the lung.S.E.H. and C.A.S. partially supported genotyping through a philanthropic donation. A.F. and D.E. were supported by a grant from the German Federal Ministry of Education and COVID-19 grant Research (BMBF; ID:01KI20197); A.F., D.E. and F.D. were supported by the Deutsche Forschungsgemeinschaft Cluster of Excellence ‘Precision Medicine in Chronic Inflammation’ (EXC2167). D.E. was supported by the German Federal Ministry of Education and Research (BMBF) within the framework of the Computational Life Sciences funding concept (CompLS grant 031L0165). D.E., K.B. and S.B. acknowledge the Novo Nordisk Foundation (NNF14CC0001 and NNF17OC0027594). T.L.L., A.T. and O.Ö. were funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation), project numbers 279645989; 433116033; 437857095. M.W. and H.E. are supported by the German Research Foundation (DFG) through the Research Training Group 1743, ‘Genes, Environment and Inflammation’. L.V. received funding from: Ricerca Finalizzata Ministero della Salute (RF-2016-02364358), Italian Ministry of Health ‘CV PREVITAL’—strategie di prevenzione primaria cardiovascolare primaria nella popolazione italiana; The European Union (EU) Programme Horizon 2020 (under grant agreement No. 777377) for the project LITMUS- and for the project ‘REVEAL’; Fondazione IRCCS Ca’ Granda ‘Ricerca corrente’, Fondazione Sviluppo Ca’ Granda ‘Liver-BIBLE’ (PR-0391), Fondazione IRCCS Ca’ Granda ‘5permille’ ‘COVID-19 Biobank’ (RC100017A). A.B. was supported by a grant from Fondazione Cariplo to Fondazione Tettamanti: ‘Bio-banking of Covid-19 patient samples to support national and international research (Covid-Bank). This research was partly funded by an MIUR grant to the Department of Medical Sciences, under the program ‘Dipartimenti di Eccellenza 2018–2022’. This study makes use of data generated by the GCAT-Genomes for Life. Cohort study of the Genomes of Catalonia, Fundació IGTP (The Institute for Health Science Research Germans Trias i Pujol) IGTP is part of the CERCA Program/Generalitat de Catalunya. GCAT is supported by Acción de Dinamización del ISCIII-MINECO and the Ministry of Health of the Generalitat of Catalunya (ADE 10/00026); the Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR) (2017-SGR 529). M.M. received research funding from grant PI19/00335 Acción Estratégica en Salud, integrated in the Spanish National RDI Plan and financed by ISCIII-Subdirección General de Evaluación and the Fondo Europeo de Desarrollo Regional (European Regional Development Fund (FEDER)-Una manera de hacer Europa’). B.C. is supported by national grants PI18/01512. X.F. is supported by the VEIS project (001-P-001647) (co-funded by the European Regional Development Fund (ERDF), ‘A way to build Europe’). Additional data included in this study were obtained in part by the COVICAT Study Group (Cohort Covid de Catalunya) supported by IsGlobal and IGTP, European Institute of Innovation & Technology (EIT), a body of the European Union, COVID-19 Rapid Response activity 73A and SR20-01024 La Caixa Foundation. A.J. and S.M. were supported by the Spanish Ministry of Economy and Competitiveness (grant numbers: PSE-010000-2006-6 and IPT-010000-2010-36). A.J. was also supported by national grant PI17/00019 from the Acción Estratégica en Salud (ISCIII) and the European Regional Development Fund (FEDER). The Basque Biobank, a hospital-related platform that also involves all Osakidetza health centres, the Basque government’s Department of Health and Onkologikoa, is operated by the Basque Foundation for Health Innovation and Research-BIOEF. M.C. received Grants BFU2016-77244-R and PID2019-107836RB-I00 funded by the Agencia Estatal de Investigación (AEI, Spain) and the European Regional Development Fund (FEDER, EU). M.R.G., J.A.H., R.G.D. and D.M.M. are supported by the ‘Spanish Ministry of Economy, Innovation and Competition, the Instituto de Salud Carlos III’ (PI19/01404, PI16/01842, PI19/00589, PI17/00535 and GLD19/00100) and by the Andalussian government (Proyectos Estratégicos-Fondos Feder PE-0451-2018, COVID-Premed, COVID GWAs). The position held by Itziar de Rojas Salarich is funded by grant FI20/00215, PFIS Contratos Predoctorales de Formación en Investigación en Salud. Enrique Calderón’s team is supported by CIBER of Epidemiology and Public Health (CIBERESP), ‘Instituto de Salud Carlos III’. J.C.H. reports grants from Research Council of Norway grant no 312780 during the conduct of the study. E.S. reports grants from Research Council of Norway grant no. 312769. The BioMaterialBank Nord is supported by the German Center for Lung Research (DZL), Airway Research Center North (ARCN). The BioMaterialBank Nord is member of popgen 2.0 network (P2N). P.K. Bergisch Gladbach, Germany and the Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, University of Cologne, Cologne, Germany. He is supported by the German Federal Ministry of Education and Research (BMBF). O.A.C. is supported by the German Federal Ministry of Research and Education and is funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy—CECAD, EXC 2030–390661388. The COMRI cohort is funded by Technical University of Munich, Munich, Germany. This work was supported by grants of the Rolf M. Schwiete Stiftung, the Saarland University, BMBF and The States of Saarland and Lower Saxony. K.U.L. is supported by the German Research Foundation (DFG, LU-1944/3-1). Genotyping for the BoSCO study is funded by the Institute of Human Genetics, University Hospital Bonn. F.H. was supported by the Bavarian State Ministry for Science and Arts. Part of the genotyping was supported by a grant to A.R. from the German Federal Ministry of Education and Research (BMBF, grant: 01ED1619A, European Alzheimer DNA BioBank, EADB) within the context of the EU Joint Programme—Neurodegenerative Disease Research (JPND). Additional funding was derived from the German Research Foundation (DFG) grant: RA 1971/6-1 to A.R. P.R. is supported by the DFG (CCGA Sequencing Centre and DFG ExC2167 PMI and by SH state funds for COVID19 research). F.T. is supported by the Clinician Scientist Program of the Deutsche Forschungsgemeinschaft Cluster of Excellence ‘Precision Medicine in Chronic Inflammation’ (EXC2167). C.L. and J.H. are supported by the German Center for Infection Research (DZIF). T.B., M.M.B., O.W. und A.H. are supported by the Stiftung Universitätsmedizin Essen. M.A.-H. was supported by Juan de la Cierva Incorporacion program, grant IJC2018-035131-I funded by MCIN/AEI/10.13039/501100011033. E.C.S. is supported by the Deutsche Forschungsgemeinschaft (DFG; SCHU 2419/2-1).Peer reviewe