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Bojkova, Denisa ; Rothenburger, Tamara ; Ciesek, Sandra ; Wass, Mark N. ; Michaelis, Martin ; Cinatl, Jindrich
Recently, we have shown that SARS-CoV-2 Omicron virus isolates are less effective at inhibiting the host cell interferon response than Delta viruses. Here, we present further evidence that reduced interferon-antagonising activity explains at least in part why Omicron variant infections are inherently less severe than infections with other SARS-CoV-2 variants. Most importantly, we here also show that Omicron variant viruses display enhanced sensitivity to interferon treatment, which makes interferons promising therapy candidates for Omicron patients, in particular in combination with other antiviral agents.
Motivation DNA CpG methylation (CpGm) has proven to be a crucial epigenetic factor in the gene regulatory system. Assessment of DNA CpG methylation values via whole-genome bisulfite sequencing (WGBS) is, however, computationally extremely demanding.
Results We present FAst MEthylation calling (FAME), the first approach to quantify CpGm values directly from bulk or single-cell WGBS reads without intermediate output files. FAME is very fast but as accurate as standard methods, which first produce BS alignment files before computing CpGm values. We present experiments on bulk and single-cell bisulfite datasets in which we show that data analysis can be significantly sped-up and help addressing the current WGBS analysis bottleneck for large-scale datasets without compromising accuracy.
Availability An implementation of FAME is open source and licensed under GPL-3.0 at https://github.com/FischerJo/FAME.
Andlauer, Till ; Guzman-Parra, José ; Streit, Fabian ; Strohmaier, Jana ; González, Maria José ; Flores, Susana Gil ; Cabaleiro Fabeiro, Francisco J. ; Rı́o Noriega, Francisco del ; Perez, Fermin Perez ; González, Jesus Haro ; Orozco Diaz, Guillermo ; de Diego-Otero, Yolanda ; Moreno-Kuestner, Berta ; Auburger, Georg ; Degenhardt, Franziska ; Heilmann-Heimbach, Stefanie ; Herms, Stefan ; Hoffmann, Per ; Frank, Josef ; Foo, Jerome Clifford ; Treutlein, Jens ; Witt, Stephanie H. ; Cichon, Sven ; Kogevinas, Manolis ; Rivas, Fabio ; Mayoral, Fermı́n ; Müller-Myhsok, Bertram ; Forstner, Andreas Josef ; Nöthen, Markus Maria ; Rietschel, Marcella
Multiplex families with a high prevalence of a psychiatric disorder are often examined to identify rare genetic variants with large effect sizes. In the present study, we analysed whether the risk for bipolar disorder (BD) in BD multiplex families is influenced by common genetic variants. Furthermore, we investigated whether this risk is conferred mainly by BD-specific risk variants or by variants also associated with the susceptibility to schizophrenia or major depression. In total, 395 individuals from 33 Andalusian BD multiplex families as well as 438 subjects from an independent, sporadic BD case-control cohort were analysed. Polygenic risk scores (PRS) for BD, schizophrenia, and major depression were calculated and compared between the cohorts. Both the familial BD cases and unaffected family members had significantly higher PRS for all three psychiatric disorders than the independent controls, suggesting a high baseline risk for several psychiatric disorders in the families. Moreover, familial BD cases showed significantly higher BD PRS than unaffected family members and sporadic BD cases. A plausible hypothesis is that, in multiplex families with a general increase in risk for psychiatric disease, BD development is attributable to a high burden of common variants that confer a specific risk for BD. The present analyses, therefore, demonstrated that common genetic risk variants for psychiatric disorders are likely to contribute to the high incidence of affective psychiatric disorders in the multiplex families. The PRS explained only part of the observed phenotypic variance and rare variants might have also contributed to disease development.
Kennedy, Eamonn ; Vadlamani, Shashank ; Lindsey, Hannah M. ; Lei, Pui-Wa ; Jo-Pugh, Mary ; Adamson, Maheen ; Alda, Martin ; Alonso-Lana, Silvia ; Ambrogi, Sonia ; Anderson, Tim J. ; Arango, Celso ; Asarnow, Robert F. ; Avram, Mihai ; Ayesa-Arriola, Rosa ; Babikian, Talin ; Banaj, Nerisa ; Bird, Laura J. ; Borgwardt, Stefan ; Brodtmann, Amy ; Brosch, Katharina ; Caeyenberghs, Karen ; Calhoun, Vince D. ; Chiaravalloti, Nancy D. ; Cifu, David X. ; Crespo-Facorro, Benedicto ; Dalrymple-Alford, John C. ; Dams-O’Connor, Kristen ; Dannlowski, Udo ; Darby, David ; Davenport, Nicholas ; DeLuca, John ; Díaz-Caneja, Covadonga M. ; Disner, Seth G. ; Dobryakova, Ekaterina ; Ehrlich, Stefan ; Esopenko, Carrie ; Ferrarelli, Fabio ; Frank, Lea E. ; Franz, Carol ; Fuentes-Claramonte, Paola ; Genova, Helen ; Giza, Christopher C. ; Goltermann, Janik ; Grotegerd, Dominik ; Gruber, Marius ; Gutiérrez-Zotes, Alfonso ; Ha, Minji ; Haavik, Jan ; Hinkin, Charles ; Hoskinson, Kristen R. ; Hubl, Daniela ; Irimia, Andrei ; Jansen, Andreas ; Kaess, Michael ; Kang, Xiaojian ; Kenney, Kimbra ; Keřková, Barbora ; Khlif, Mohamed Salah ; Kim, Minah ; Kindler, Jochen ; Kircher, Tilo ; Knı́žková, Karolina ; Kolskår, Knut K. ; Krch, Denise ; Kremen, William S. ; Kuhn, Taylor ; Kumari, Veena ; Kwon, Jun Soo ; Langella, Roberto ; Laskowitz, Sarah ; Lee, Jungha ; Lengenfelder, Jean ; Liebel, Spencer W. ; Liou-Johnson, Victoria ; Lippa, Sara M. ; Løvstad, Marianne ; Lundervold, Astri ; Marotta, Cassandra ; Marquardt, Craig A. ; Mattos, Paulo ; Mayeli, Ahmad ; McDonald, Carrie R. ; Meinert, Susanne ; Melzer, Tracy R. ; Merchán Naranjo, Jessica ; Michel, Chantal ; Morey, Rajendra A. ; Mwangi, Benson ; Myall, Daniel J. ; Nenadić, Igor ; Newsome, Mary R. ; Nunes, Abraham ; O’Brien, Terence J. ; Oertel, Viola ; Ollinger, John ; Olsen, Alexander ; Ortiz Garcı́a de la Foz, Victor ; Ozmen, Mustafa ; Pardoe, Heath ; Parent, Marise ; Piras, Fabrizio ; Piras, Federica ; Pomarol-Clotet, Edith ; Repple, Jonathan ; Richard, Geneviève ; Rodriguez, Jonathan ; Rodriguez, Mabel ; Rootes-Murdy, Kelly ; Rowland, Jared ; Ryan, Nicholas P. ; Salvador, Raymond ; Sanders, Anne-Marthe ; Schmidt, André ; Soares, Jair C. ; Spalleta, Gianfranco ; Španiel, Filip ; Stasenko, Alena ; Stein, Frederike ; Straube, Benjamin ; Thames, April ; Thomas-Odenthal, Florian ; Thomopoulos, Sophia I. ; Tone, Erin ; Torres, Ivan ; Troyanskaya, Maya ; Turner, Jessica A. ; Ulrichsen, Kristine M. ; Umpierrez, Guillermo ; Vilella, Elisabet ; Vivash, Lucy ; Walker, William C. ; Werden, Emilio ; Westlye, Lars T. ; Wild, Krista ; Wroblewski, Adrian ; Wu, Mon-Ju ; Wylie, Glenn R. ; Yatham, Lakshmi N. ; Zunta-Soares, Giovana B. ; Thompson, Paul M. ; Tate, David F. ; Hillary, Frank G. ; Dennis, Emily L. ; Wilde, Elisabeth A.
Investigators in the cognitive neurosciences have turned to Big Data to address persistent replication and reliability issues by increasing sample sizes, statistical power, and representativeness of data. While there is tremendous potential to advance science through open data sharing, these efforts unveil a host of new questions about how to integrate data arising from distinct sources and instruments. We focus on the most frequently assessed area of cognition - memory testing - and demonstrate a process for reliable data harmonization across three common measures. We aggregated raw data from 53 studies from around the world which measured at least one of three distinct verbal learning tasks, totaling N = 10,505 healthy and brain-injured individuals. A mega analysis was conducted using empirical bayes harmonization to isolate and remove site effects, followed by linear models which adjusted for common covariates. After corrections, a continuous item response theory (IRT) model estimated each individual subject’s latent verbal learning ability while accounting for item difficulties. Harmonization significantly reduced inter-site variance by 37% while preserving covariate effects. The effects of age, sex, and education on scores were found to be highly consistent across memory tests. IRT methods for equating scores across AVLTs agreed with held-out data of dually-administered tests, and these tools are made available for free online. This work demonstrates that large-scale data sharing and harmonization initiatives can offer opportunities to address reproducibility and integration challenges across the behavioral sciences.
Mapping cortical brain asymmetry in 17,141 healthy individuals worldwide via the ENIGMA Consortium
(2017)
Kong, Xiang-Zhen ; Mathias, Samuel R. ; Guadalupe, Tulio ; Abé, Christoph ; Agartz, Ingrid ; Akudjedu, Theophilus N. ; Andre, Aleman ; Saud, Alhusaini ; Allen, Nicholas B. ; Ames, David ; Andreassen, Ole A. ; Arias Vásquez, Alejandro ; Armstrong, Nicola J. ; Bergo, Felipe ; Bastin, Mark E. ; Batalla, Albert ; Bauer, Jochen ; Baune, Bernhard T. ; Baur, Ramona ; Biederman, Joseph ; Blaine, Sara K. ; Boedhoe, Premika ; Bøen, Erlend ; Bose, Anushree ; Bralten, Janita ; Brandeis, Daniel ; Brem, Silvia ; Brodaty, Henry ; Bröhl, Henrieke ; Brooks, Samantha J. ; Buitelaar, Jan K. ; Bürger, Christian ; Bülow, Robin ; Calhoun, Vince D. ; Calvo, Anna ; Canales-Rodrı́guez, Erick Jorge ; Canive, Jose M. ; Cannon, Dara M. ; Caparelli, Elisabeth C. ; Castellanos, Francisco X. ; Cavalleri, Gianpiero L. ; Cendes, Fernando ; Chaim-Avancini, Tiffany Moukbel ; Chantiluke, Kaylita ; Chen, Qun-lin ; Chen, Xiayu ; Cheng, Yuqi ; Christakou, Anastasia ; Clark, Vincent P. ; Coghill, David ; Connolly, Colm G. ; Conzelmann, Annette ; Cόrdova-Palomera, Aldo ; Cousijn, Janna ; Crow, Tim ; Cubillo, Ana ; Dannlowski, Udo ; de Bruttopilo, Sara Ambrosino ; Zeeuw, Patrick de ; Deary, Ian J. ; Delanty, Norman ; Demeter, Damion V. ; Di Martino, Adriana ; Dickie, Erin W ; Dietsche, Bruno ; Doan, N. Trung ; Doherty, Colin P. ; Doyle, Alysa ; Durston, Sarah ; Earl, Eric ; Ehrlich, Stefan ; Ekman, Carl Johan ; Elvsåshagen, Torbjørn ; Epstein, Jeffery N. ; Fair, Damien A. ; Faraone, Stephen V. ; Fatouros-Bergman, Helena ; Fernndez, Guillen ; Busatto Filho, Geraldo ; Flyckt, Lena ; Forster, Katharina ; Jean-Paul, Fouche ; Foxe, John J. ; Fuentes-Claramonte, Paola ; Fullerton, Janice M. ; Garavan, Hugh ; Garcia, Danielle do Santos ; Gotlib, Ian H. ; Goudriaan, Anna E. ; Grabe, Hans Jörgen ; Groenewold, Nynke A. ; Grotegerd, Dominik ; Gruber, Oliver ; Gurholt, Tiril ; Haavik, Jan ; Hahn, Tim ; Hansell, Narelle K. ; Harris, Mathew A. ; Hartman, Catharina ; Hernandez, Maria del Carmen Valdes ; Heslenfeld, Dirk ; Hester, Robert ; Hibar, Derrek Paul ; Ho, Beng-Choon ; Ho, Tiffany C. ; Hoekstra, Pieter J. ; van Holst, Ruth J. ; Hoogman, Martine ; Hovik, Marie F. ; Howells, Fleur M. ; Hugdahl, Kenneth ; Huyser, Chaim ; Ingvar, Martind ; Irwin, Lourdes ; Ishikawa, Akari ; James, Anthony ; Jahanshad, Neda ; Jernigan, Terry L. ; Jönsson, Erik G. ; Kahler, Claas ; Kaleda, Vasily ; Kelly, Clare ; Kerich, Michael ; Keshavan, Matcheri S ; Khadka, Sabin ; Kircher, Tilo ; Kohls, Gregor ; Konrad, Kerstin ; Korucuoglu, Ozlem ; Krämer, Bernd ; Krug, Axel ; Kwon, Jun Soo ; Lambregts-Rommelse, Nanda ; Landén, Mikael ; Lázaro, Luisa ; Lebedeva, Irina ; Lenroot, Rhoshel ; Lesch, Klaus-Peter J. ; Li, Qinqin ; Lim, Kelvin O. ; Liu, Jia ; Lochner, Christine ; London, Edythe D. ; Lonning, Vera ; Lorenzetti, Valentina ; Luciano, Michelle ; Luijten, Maartje ; Lundervold, Astri J. ; Mackey, Scott ; MacMaster, Frank P. ; Maingault, Sophie ; Malpas, Charles B. ; Malt, Ulrik F. ; Mataix-Cols, David ; Martin-Santos, Rocio ; Mayer, Andrew R. ; McCarthy, Hazel ; Mitchell, Philip B. ; Mueller, Bryon A. ; Maniega, Susana Munoz ; Mazoyer, Bernard ; McDonald, Colm ; McLellan, Quinn ; McMahon, Katie L. ; McPhilemy, Genevieve ; Momenan, Reza ; Morales, Angelica M. ; Narayanaswamy, Janardhanan C. ; Moreira, José Carlos Vasques ; Nerland, Stener ; Nestor, Liam ; Nigg, Joel T. ; Nordvik, Jan-Egil ; Novotny, Stephanie ; Oberwelland, Eileen ; O’Gorman, Ruth L. ; Oosterlaan, Jaap ; Oranje, Bob ; Orr, Catherine ; Overs, Bronwyn ; Pauli, Paul ; Paulus, Martin ; Plessen, Kerstin ; Polier, Georg von ; Pomarol-Clotet, Edith ; Qiu, Jiang ; Radua, Joaquim ; Ramos-Quiroga, J. Antoni ; Reddy, Y.C. Janardhan ; Reif, Andreas ; Roberts, Gloria ; Rosa, Pedro ; Rubia, Katya ; Sacchet, Matthew D. ; Sachdev, Perminder S. ; Salvador, Raymond ; Schmaal, Lianne ; Schweren, Lisanne ; Seidman, Larry J. ; Seitz, Jochen ; Serpa, Mauricio Henriques ; Shaw, Philip ; Shumskaya, Elena ; Silk, Timothy J. ; Simmons, Alan N. ; Simulionyte, Egle ; Sinha, Rajita ; Sjoerds, Zsuzsika ; Smelror, Runar Elle ; Soliva, Joan Carlos ; Solowij, Nadia ; Sponheim, Scott R. ; Stein, Dan J. ; Stein, Elliot A. ; Stevens, Michael ; Strike, Lachlan T. ; Sudre, Gustavo ; Sui, Jing ; Tamm, Leanne ; Temmingh, Hendrik S. ; Thoma, Robert J. ; Tomyshev, Alexander S. ; Tronchin, Giulia ; Turner, Jessica A. ; Uhlmann, Anne ; van Erp, Theo G.M. ; van den Heuvel, Odile ; Meer, Dennis van der ; Eijk, Liza van ; Vance, Alasdair ; Veer, Ilya M. ; Veltman, Dick J. ; Venkatasubramanian, Ganesan ; Vilarroya, Oscar ; Vives-Gilabert, Yolanda ; Voineskos, Aristotle N. ; Völzke, Henry ; Vuletic, Daniella ; Walitza, Susanne ; Walter, Henrik ; Walton, Esther ; Wardlaw, Joanna M. ; Wen, Wei ; Westlye, Lars T. ; Whelan, Christopher D. ; White, Tonya ; Wiers, Reinout W. ; Wright, Margaret J. ; Wittfeld, Katharina ; Yang, Tony T. ; Yasuda, Clarissa L. ; Yoncheva, Yuliya ; Yucel, Murat ; Yun, Je-Yeon ; Zanetti, Marcus Vinicius ; Zhen, Zonglei ; Zhu, Xing-xing ; Ziegler, Georg Christoph ; Zierhut, Kathrin ; De Zubicaray, Greig ; Zwiers, Marcel P. ; Glahn, David C. ; Franke, Barbara ; Crivello, Fabrice ; Tzourio-Mazoyer, Nathalie ; Fisher, Simon E. ; Thompson, Paul M. ; Francks, Clyde ; Farde, Lars ; Engberg, Goran ; Erhardt, Sophie ; Cervenka, Simon ; Schwieler, Lilly ; Piehl, Fredrik ; Collste, Karin ; Victorsson, Pauliina ; Malmqvist, Anna ; Hedberg, Mikael ; Orhan, Funda
Hou, Liping ; Bergen, Sarah E. ; Akula, Nirmala ; Song, Jie ; Hultman, Christina M. ; Landén, Mikael ; Adli, Mazda ; Alda, Martin ; Ardau, Raffaella ; Arias, Bárbara ; Aubry, Jean-Michel ; Backlund, Lena ; Badner, Judith A. ; Barrett, Thomas B. ; Bauer, Michael ; Baune, Bernhard T. ; Bellivier, Frank ; Benabarre Hernandez, Antonio ; Bengesser, Susanne ; Berrettini, Wade H. ; Bhattacharjee, Abesh Kumar ; Biernacka, Joanna ; Birner, Armin ; Bloss, Cinnamon S. ; Brichant-Petitjean, Clara ; Bui, Elise T. ; Byerley, William ; Cervantes, Pablo ; Chillotti, Caterina ; Cichon, Sven ; Colom, Francesc ; Coryell, William ; Craig, David W. ; Cruceanu, Cristiana ; Czerski, Piotr M. ; Davis, Tony ; Dayer, Alexandre ; Degenhardt, Franziska ; Del Zompo, Maria ; DePaulo, J. Raymond ; Edenberg, Howard J. ; Étain, Bruno ; Falkai, Peter ; Foroud, Tatiana ; Forstner, Andreas Josef ; Frisén, Louise ; Frye, Mark A. ; Fullerton, Janice M. ; Gard, Sébastien ; Garnham, Julie S. ; Gershon, Elliot S. ; Goes, Fernando S. ; Greenwood, Tiffany A. ; Grigoroiu-Serbanescu, Maria ; Hauser, Joanna ; Heilbronner, Urs ; Heilmann-Heimbach, Stefanie ; Herms, Stefan ; Hipolito, Maria ; Hitturlingappa, Shashi ; Hoffmann, Per ; Hofmann, Andrea ; Jamain, Stephane ; Jiménez, Esther ; Kahn, Jean-Pierre ; Kassem, Layla ; Kelsoe, John R. ; Kittel-Schneider, Sarah ; Kliwicki, Sebastian ; Koller, Daniel L. ; König, Barbara ; Lackner, Nina ; Laje, Gonzalo ; Lang, Maren ; Lavebratt, Catharina ; Lawson, William B. ; Leboyer, Marion ; Leckband, Susan G. ; Liu, Chunyu ; Maaser, Anna ; Mahon, Pamela B. ; Maier, Wolfgang ; Maj, Mario ; Manchia, Mirko ; Martinsson, Lina ; McCarthy, Michael J. ; McElroy, Susan L. ; McInnis, Melvin G. ; McKinney, Rebecca ; Mitchell, Philip B. ; Mitjans, Marina ; Mondimore, Francis Mark ; Monteleone, Palmiero ; Mühleisen, Thomas W. ; Nievergelt, Caroline M. ; Nöthen, Markus Maria ; Novák, Tomas ; Nurnberger, John I. ; Nwulia, Evaristus A. ; Ösby, Urban ; Pfennig, Andrea ; Potash, James B. ; Propping, Peter ; Reif, Andreas ; Reininghaus, Eva ; Rice, John ; Rietschel, Marcella ; Rouleau, Guy A. ; Rybakowski, Janusz K. ; Schalling, Martin ; Scheftner, William A. ; Schofield, Peter R. ; Schork, Nicholas J. ; Schulze, Thomas Gerd ; Schumacher, Johannes ; Schweizer, Barbara W. ; Severino, Giovanni ; Shekhtman, Tatyana ; Shilling, Paul D. ; Simhandl, Christian ; Slaney, Claire M. ; Smith, Erin N. ; Squassina, Alessio ; Stamm, Thomas ; Stopkova, Pavla ; Streit, Fabian ; Strohmaier, Jana ; Szelinger, Szabolcs ; Tighe, Sarah K. ; Tortorella, Alfonso ; Turecki, Gustavo ; Vieta, Eduard ; Volkert, Julia ; Witt, Stephanie H. ; Wright, Adam ; Zandi, Peter P. ; Zhang, Peng ; Zollner, Sebastian ; McMahon, Francis J.
Bipolar disorder (BD) is a genetically complex mental illness characterized by severe oscillations of mood and behavior. Genome-wide association studies (GWAS) have identified several risk loci that together account for a small portion of the heritability. To identify additional risk loci, we performed a two-stage meta-analysis of >9 million genetic variants in 9,784 bipolar disorder patients and 30,471 controls, the largest GWAS of BD to date. In this study, to increase power we used ~2,000 lithium-treated cases with a long-term diagnosis of BD from the Consortium on Lithium Genetics, excess controls, and analytic methods optimized for markers on the Xchromosome. In addition to four known loci, results revealed genome-wide significant associations at two novel loci: an intergenic region on 9p21.3 (rs12553324, p = 5.87×10-9; odds ratio = 1.12) and markers within ERBB2 (rs2517959, p = 4.53×10-9; odds ratio = 1.13). No significant X-chromosome associations were detected and X-linked markers explained very little BD heritability. The results add to a growing list of common autosomal variants involved in BD and illustrate the power of comparing well-characterized cases to an excess of controls in GWAS.
Der Zusammenhang zwischen kognitiver Emotionsregulation, positivem Aufmerksamkeitsbias und Resilienz
(2022)
Das Verständnis von Faktoren, die die Widerstandsfähigkeit gegen Stress fördern, ist entscheidend für die Entwicklung von Stresspräventionsprogrammen und für die Verbesserung der Behandlung stressbedingter Störungen. Zum einen war es Ziel der vorliegenden Forschungsarbeit, den Einfluss der kognitiven Emotionsregulation (ER) auf die psychische Gesundheit zu untersuchen und bereits vorhandene Ergebnisse zum Einfluss der Emotionsregulation auf die Resilienz zu replizieren. Es wird zunehmend anerkannt, dass die meisten psychiatrischen Erkrankungen mit Emotionsdysregulation einhergehen und dass klinische Interventionen davon profitieren, wenn sie auf einem empirischen Verständnis der Emotionsprozesse beruhen.
Der Hauptfokus lag zudem darauf, zu untersuchen, ob es einen Zusammenhang zwischen Resilienz und einem Aufmerksamkeitsbias auf positive Informationen, einen Positivitätsbias, gibt.
In der vorliegenden Studie wurde eine Stichprobe psychisch gesunder Teilnehmer (n=229) im Alter von 18 bis 55 Jahren herangezogen, denen Bilder aus dem „International Affective Picture System“ präsentiert wurden. Dabei bekamen sie die Anweisung, negative Emotionen zu Fotos durch kognitives Umbewerten oder Distanzieren herunterzuregulieren. Die Reaktionszeiten sowie die Erregungsbewertungen wurden für die Bedingungen Neubewertung, Distanzieren, negatives passives Betrachten sowie neutrales passives Betrachten erhoben. Zudem wurde die Aufmerksamkeitsverzerrung gegenüber positiven und negativen Reizen mithilfe einer Visual Dot-Probe Aufgabe untersucht. Gemessen wurden die Reaktionszeiten der Studienteilnehmer bei Reaktionen auf einen Stimulus, der auf die Präsentation emotionaler Gesichter folgt, im Vergleich zu Reaktionen auf einen Stimulus, der an die Stelle von neutralen Gesichtern rückt. Hieraus wurden Aufmerksamkeitsverzerrungen abgeleitet.
Die anschließende Datenanalyse und statistische Auswertung konnten zeigen, dass die Neubewertung im Vergleich zum passiven Betrachten der negativen Bilder eine längere Reaktionszeit aufweist, was darauf hinweist, dass die kognitive Emotionsregulation möglicherweise anstrengendere kognitive Kontrollprozesse aktiviert. Hinsichtlich des Zusammenhangs mit der Resilienz konnte eine signifikante positive Korrelation der Reaktionszeitdifferenzen der Emotionsregulationsstrategien Distanzieren und Neubewerten mit dem Resilienz-Score beobachtet werden.
Bei den Erregungsbewertungen der Emotionsregulation zeigte sich weder ein signifikanter Effekt der Bedingung noch ein Zusammenhang mit dem Resilienz-Score.
Die Ergebnisse der Visual Dot-Probe zeigten, dass es keine Unterschiede in den Reaktionszeiten nach der Präsentation des neutralen sowie des emotionalen Stimulus gab. Auch zeigten sich keine Zusammenhänge mit der Resilienz.
Der erwartete Zusammenhang zwischen Resilienz und einem Aufmerksamkeitsbias auf positive Informationen konnte in den Ergebnissen nicht gefunden werden. In den vorliegenden Daten zeigte sich hingegen eine signifikante negative Korrelation der Fähigkeit der kognitiven Neubewertung mit der Höhe des negativen Aufmerksamkeits Bias-Scores.
Somit liefert die vorliegende Arbeit einen Hinweis darauf, dass Personen mit einem hohen Maß an Emotionsregulationskapazitäten eine höhere Aufmerksamkeitslenkung weg von emotional negativen Stimuli aufweisen.
Bezüglich der Anwendung auf klinische Stichproben besteht noch Raum zu
analysieren, ob es sich um ein stabiles Phänomen handelt, das auf diese übertragen werden kann.
Der Nutzen von DJ-Kathetern ist unverzichtbar für die moderne Urologie. Sie sind essenziell, um bei einer Vielzahl von Erkrankungen einen adäquaten Harnabfluss und dementsprechend eine gute Nierenfunktion gewährleisten zu können. Dabei können sie für eine temporäre oder dauerhafte Schienung genutzt werden. Als einliegende Fremdkörper können sie, insbesondere bei langfristiger Nutzung, leicht durch Bakterien und Pilze kolonisiert werden und beherbergen somit ein erhöhtes Risiko für Harnwegsinfekte.
Das Hauptziel dieser Arbeit ist folglich, durch das Auffinden von protektiven oder prädisponierenden Faktoren für die Entwicklung von fieberhaften Harnwegsinfekten in der Zukunft Mortalität, Resistenzbildung und Kosten sowohl bei DJ-Katheter-Dauerversorgung als auch generell senken zu können. Zusätzlich soll untersucht werden, ob verschiedene Antibiotika-Regime einen Einfluss auf die Entwicklung postoperativer, fieberhafter Harnwegsinfekte haben.
Um dies feststellen zu können, wurden in dieser Studie 100 Patientinnen und Patienten eingeschlossen, die von 2013 bis 2018 in der Klinik für Urologie des Universitätsklinikums Frankfurt zum DJ-Wechsel bei DJ-Katheter-Dauerversorgung vorstellig wurden. Anschließend wurden verschiedene Faktoren untersucht, die das Risiko für das Auftreten von fieberhaften Harnwegsinfekten oder die Entwicklung von Resistenzen in Krankheitserregern erhöhen oder reduzieren könnten.
Hierzu wurden insgesamt 950 DJ-Katheter-assoziierte Eingriffe (Einlage, Wechsel, Entfernung) analysiert. Der individuelle Beobachtungszeitraum betrug durchschnittlich 2,9 Jahre mit durchschnittlich 7 DJ-Katheter-Wechsel.
Gegenüber der Normalbevölkerung wies die Studienpopulation, am ehesten durch die einliegenden DJ-Katheter, eine deutlich erhöhte Prävalenz von Harnwegsinfekten auf (18,53% vs. 2,5%). Darüber hinaus konnte gezeigt werden, dass ein höheres Patientenalter mit einem gering erhöhten Risiko für die Ausbildung von resistenten Keimen im Urin korreliert (p=0,0121; OR 1,0395; KI 1,0096-1,0731 (univariate Analyse), p=0,0030; OR 1,0618 KI 1,0226-1,1077 (multivariate Analyse)). Dies korrelierte jedoch nicht mit einem erhöhten Risiko für fieberhafte Harnwegsinfekte. Darüber hinaus konnte festgestellt werden, dass die operative Manipulation unter empirischer oder testgerechter, antibiotischer Therapie nicht mit einem hohen Risiko für die Entstehung fieberhafter Harnwegsinfekte einhergeht. In den analysierten Daten fanden sich insgesamt zu wenige fieberhafte Infekte (n=72; 7,58%), um das Ziel begünstigender oder protektiver Faktoren für die Entstehung von fieberhaften Harnwegsinfekten adäquat zu untersuchen. Aus dem gleichen Grund konnten aus den Daten auch keine Hinweise für das optimale Antibiotika-Regime abgeleitet werden. In fast 70% der untersuchten Fälle wurde eine periinterventionelle single-shot Antibiose angewendet, weswegen diese als Infektionsprophylaxe einen adäquaten Schutz darzustellen scheint.
Generell kann davon ausgegangen werden, dass die DJ-Katheter-Dauerversorgung, trotz der möglichen mikrobiellen Besiedlung mit eventuellen Komplikationen wie Bakteriurie oder systemischen Infektionen, ein sicheres Verfahren ist. Manipulationen oder operativer Wechsel der DJ-Katheter stellen, trotz Präsenz der Keime, keine riskanten Manöver dar, sofern diese adäquat und unter entsprechender, antibiotischer Therapie durchgeführt werden.
Weitere, noch größere und insbesondere prospektive, randomisierte Studien sind zu empfehlen und könnten die Ergebnisse weiter bestätigen und erweitern, vor allem in Bezug auf die Überlegenheit verschiedener antibiotischer Regime in der Vermeidung einer Resistenzbildung.
Damme, Ellen van ; De Meyer, Sandra ; Bojkova, Denisa ; Ciesek, Sandra ; Cinatl, Jindrich ; De Jonghe, Steven ; Jochmans, Dirk ; Leyssen, Pieter ; Buyck, Christophe ; Neyts, Johan ; Loock, Marnix van
Although vaccination campaigns are currently being rolled out to prevent coronavirus disease (COVID-19), antivirals will remain an important adjunct to vaccination. Antivirals against coronaviruses do not exist, hence global drug repurposing efforts have been carried out to identify agents that may provide clinical benefit to patients with COVID-19. Itraconazole, an antifungal agent, has been reported to have activity against animal coronaviruses. Using cell-based phenotypic assays, the in vitro antiviral activity of itraconazole and 17-OH itraconazole was assessed against clinical isolates from a German and Belgian patient infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Itraconazole demonstrated antiviral activity in human Caco-2 cells (EC50 = 2.3 µM; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay). Similarly, its primary metabolite, 17-OH itraconazole, showed inhibition of SARS-CoV-2 activity (EC50 = 3.6 µM). Remdesivir inhibited viral replication with an EC50 = 0.4 µM. Itraconazole and 17-OH itraconazole resulted in a viral yield reduction in vitro of approximately 2-log10 and approximately 1-log10, as measured in both Caco-2 cells and VeroE6-eGFP cells, respectively. The viral yield reduction brought about by remdesivir or GS-441524 (parent nucleoside of the antiviral prodrug remdesivir; positive control) was more pronounced, with an approximately 3-log10 drop and >4-log10 drop in Caco-2 cells and VeroE6-eGFP cells, respectively. Itraconazole and 17-OH itraconazole exert in vitro low micromolar activity against SARS-CoV-2. Despite the in vitro antiviral activity, itraconazole did not result in a beneficial effect in hospitalized COVID-19 patients in a clinical study (EudraCT Number: 2020-001243-15).
Strien, Joeri van ; Haupt, Alexander ; Schulte, Uwe ; Braun, Hans-Peter ; Cabrera-Orefice, Alfredo ; Choudhary, Jyoti S. ; Evers, Felix ; Fernandez-Vizarra, Erika ; Guerrero-Castillo, Sergio ; Kooij, Taco W.A. ; Páleníková, Petra ; Pardo, Mercedes ; Ugalde, Cristina ; Wittig, Ilka ; Wöhlbrand, Lars ; Brandt, Ulrich ; Arnold, Susanne ; Huynen, Martijn A.
Complexome profiling is an emerging ‘omics’ approach that systematically interrogates the composition of protein complexes (the complexome) of a sample, by combining biochemical separation of native protein complexes with mass-spectrometry based quantitation proteomics. The resulting fractionation profiles hold comprehensive information on the abundance and composition of the complexome, and have a high potential for reuse by experimental and computational researchers. However, the lack of a central resource that provides access to these data, reported with adequate descriptions and an analysis tool, has limited their reuse. Therefore, we established the ComplexomE profiling DAta Resource (CEDAR, www3.cmbi.umcn.nl/cedar/), an openly accessible database for depositing and exploring mass spectrometry data from complexome profiling studies. Compatibility and reusability of the data is ensured by a standardized data and reporting format containing the “minimum information required for a complexome profiling experiment” (MIACE). The data can be accessed through a user-friendly web interface, as well as programmatically using the REST API portal. Additionally, all complexome profiles available on CEDAR can be inspected directly on the website with the profile viewer tool that allows the detection of correlated profiles and inference of potential complexes. In conclusion, CEDAR is a unique, growing and invaluable resource for the study of protein complex composition and dynamics across biological systems.
Immune-mediated inflammatory diseases (IMIDs), such as rheumatoid arthritis (RA), psoriatic arthritis (PsA), and psoriasis (Ps), represent autoinflammatory and autoimmune disorders, as well as conditions that have an overlap of both categories. Understanding the underlying pathogeneses, making diagnoses, and choosing individualized treatments remain challenging due to heterogeneous disease phenotypes and the lack of reliable biomarkers that drive the treatment choice. In this review, we provide an overview of the low-molecular-weight metabolites that might be employed as biomarkers for various applications, e.g., early diagnosis, disease activity monitoring, and treatment-response prediction, in RA, PsA, and Ps. The literature was evaluated, and putative biomarkers in different matrices were identified, categorized, and summarized. While some of these candidate biomarkers appeared to be disease-specific, others were shared across multiple IMIDs, indicating common underlying disease mechanisms. However, there is still a long way to go for their application in a routine clinical setting. We propose that studies integrating omics analyses of large patient cohorts from different IMIDs should be performed to further elucidate their pathomechanisms and treatment options. This could lead to the identification and validation of biomarkers that might be applied in the context of precision medicine to improve the clinical outcomes of these IMID patients.
Zuraw, Bruce L. ; Lumry, William R. ; Johnston, Douglas T. ; Aygören-Pürsün, Emel ; Banerji, Aleena ; Bernstein, Jonathan A. ; Christiansen, Sandra C. ; Jacobs, Joshua S. ; Sitz, Karl V. ; Gower, Richard G. ; Gagnon, Remi ; Wedner, H. James ; Kinaciyan, Tamar ; Hakl, Roman ; Hanzlíková, Jana ; Anderson, John T. ; McNeil, Donald L. ; Fritz, Stephen B. ; Yang, William H. ; Tachdjian, Raffi ; Busse, Paula J. ; Craig, Timothy J. ; Li, H. Henry ; Farkas, Henriette ; Best, Jessica M. ; Clemons, Desiree ; Cornpropst, Melanie ; Dobo, Sylvia M. ; Iocca, Heather A. ; Kargl, Deborah ; Nagy, Eniko ; Murray, Sharon C. ; Collis, Phil ; Sheridan, William P. ; Maurer, Marcus ; Riedl, Marc A.
Background: Berotralstat (BCX7353) is an oral, once-daily inhibitor of plasma kallikrein in development for the prophylaxis of hereditary angioedema (HAE) attacks.
Objective: Our aim was to determine the efficacy, safety, and tolerability of berotralstat in patients with HAE over a 24-week treatment period (the phase 3 APeX-2 trial).
Methods: APeX-2 was a double-blind, parallel-group study that randomized patients at 40 sites in 11 countries 1:1:1 to receive once-daily berotralstat in a dose of 110 mg or 150 mg or placebo (Clinicaltrials.gov identifier NCT03485911). Patients aged 12 years or older with HAE due to C1 inhibitor deficiency and at least 2 investigator-confirmed HAE attacks in the first 56 days of a prospective run-in period were eligible. The primary efficacy end point was the rate of investigator-confirmed HAE attacks during the 24-week treatment period.
Results: A total of 121 patients were randomized; 120 of them received at least 1 dose of the study drug (n = 41, 40, and 39 in the 110-mg dose of berotralstat, 150-mg of dose berotralstat, and placebo groups, respectively). Berotralstat demonstrated a significant reduction in attack rate at both 110 mg (1.65 attacks per month; P = .024) and 150 mg (1.31 attacks per month; P < .001) relative to placebo (2.35 attacks per month). The most frequent treatment-emergent adverse events that occurred more with berotralstat than with placebo were abdominal pain, vomiting, diarrhea, and back pain. No drug-related serious treatment-emergent adverse events occurred.
Conclusion: Both the 110-mg and 150-mg doses of berotralstat reduced HAE attack rates compared with placebo and were safe and generally well tolerated. The most favorable benefit-to-risk profile was observed at a dose of 150 mg per day.
Voigt, Gesche M. ; Thiele, Dominik ; Wetzke, Martin ; Weidemann, Jürgen ; Parpatt, Patricia‐Maria ; Welte, Tobias ; Seidenberg, Jürgen ; Vogelberg, Christian ; Köster, Holger ; Rohde, Gernot Gerhard Ulrich ; Härtel, Christoph ; Hansen, Gesine ; Kopp, Matthias
Although chest radiograph (CXR) is commonly used in diagnosing pediatric community acquired pneumonia (pCAP), limited data on interobserver agreement among radiologists exist. PedCAPNETZ is a prospective, observational, and multicenter study on pCAP. N = 233 CXR from patients with clinical diagnosis of pCAP were retrieved and n = 12 CXR without pathological findings were added. All CXR were interpreted by a radiologist at the site of recruitment and by two external, blinded pediatric radiologists. To evaluate interobserver agreement, the reporting of presence or absence of pCAP in CXR was analyzed, and prevalence and bias-adjusted kappa (PABAK) statistical testing was applied. Overall, n = 190 (82%) of CXR were confirmed as pCAP by two external pediatric radiologists. Compared with patients with pCAP negative CXR, patients with CXR-confirmed pCAP displayed higher C-reactive protein levels and a longer duration of symptoms before enrollment (p < .007). Further parameters, that is, age, respiratory rate, and oxygen saturation showed no significant difference. The interobserver agreement between the onsite radiologists and each of the two independent pediatric radiologists for the presence of pCAP was poor to fair (69%; PABAK = 0.39% and 76%; PABAK = 0.53, respectively). The concordance between the external radiologists was fair (81%; PABAK = 0.62). With regard to typical CXR findings for pCAP, chance corrected interrater agreement was highest for pleural effusions, infiltrates, and consolidations and lowest for interstitial patterns and peribronchial thickening. Our data show a poor interobserver agreement in the CXR-based diagnosis of pCAP and emphasized the need for harmonized interpretation standards.
Fang, Pan ; Ji, Yanlong ; Silbern, Ivan ; Viner, Rosa ; Oellerich, Thomas ; Pan, Kuan-Ting ; Urlaub, Henning
The heterogeneity and complexity of glycosylation hinder the depth of site-specific glycoproteomics analysis. High-field asymmetric-waveform ion-mobility spectrometry (FAIMS) has been shown to improve the scope of bottom-up proteomics. The benefits of FAIMS for quantitative N-glycoproteomics have not been investigated yet. In this work, we optimized FAIMS settings for N-glycopeptide identification, with or without the tandem mass tag (TMT) label. The optimized FAIMS approach significantly increased the identification of site-specific N-glycopeptides derived from the purified immunoglobulin M (IgM) protein or human lymphoma cells. We explored in detail the changes in FAIMS mobility caused by N-glycopeptides with different characteristics, including TMT labeling, charge state, glycan type, peptide sequence, glycan size, and precursor m/z. Importantly, FAIMS also improved multiplexed N-glycopeptide quantification, both with the standard MS2 acquisition method and with our recently developed Glyco-SPS-MS3 method. The combination of FAIMS and Glyco-SPS-MS3 methods provided the highest quantitative accuracy and precision. Our results demonstrate the advantages of FAIMS for improved mass spectrometry-based qualitative and quantitative N-glycoproteomics.
The hexosamine biosynthetic pathway (HBP) is essential for the production of uridine diphosphate N-acetylglucosamine (UDP-GlcNAc), the building block of glycosaminoglycans, thus playing a crucial role in cartilage anabolism. Although O-GlcNAcylation represents a protective regulatory mechanism in cellular processes, it has been associated with degenerative diseases, including osteoarthritis (OA). The present study focuses on HBP-related processes as potential therapeutic targets after cartilage trauma. Human cartilage explants were traumatized and treated with GlcNAc or glucosamine sulfate (GS); PUGNAc, an inhibitor of O-GlcNAcase; or azaserine (AZA), an inhibitor of GFAT-1. After 7 days, cell viability and gene expression analysis of anabolic and catabolic markers, as well as HBP-related enzymes, were performed. Moreover, expression of catabolic enzymes and type II collagen (COL2) biosynthesis were determined. Proteoglycan content was assessed after 14 days. Cartilage trauma led to a dysbalanced expression of different HBP-related enzymes, comparable to the situation in highly degenerated tissue. While GlcNAc and PUGNAc resulted in significant cell protection after trauma, only PUGNAc increased COL2 biosynthesis. Moreover, PUGNAc and both glucosamine derivatives had anti-catabolic effects. In contrast, AZA increased catabolic processes. Overall, “fueling” the HBP by means of glucosamine derivatives or inhibition of deglycosylation turned out as cells and chondroprotectives after cartilage trauma.
Aufgrund einer Vielzahl an Patienten mit sekundären Lebertumoren sind aus medizinischen oder technischen Gründen nicht operabel sind, erfahren alternative lokalablative Behandlungsformen in den letzten beiden Jahrzehnten zunehmende Verbreitung.
Das Ziel der hier vorliegenden Studie ist es, das Therapieansprechen und die Sicherheit von Mikrowellen- (MWA) und Radiofrequenzablation (RFA) von Lebermetastasen in einer randomisierten Studie zu bestimmen und prospektiv zu vergleichen.
Es unterzogen sich 50 Patienten (27 Frauen, 23 Männer, mittleres Alter 62,8 Jahre) einer CT-gesteuerten perkutanen thermischen Ablation von 50 Lebermetastasen. Davon erhielten 26 Patienten eine MWA und 24 Patienten eine Behandlung mit RFA. Das kolorektale Karzinom war in 54 % der Fälle ein Primärtumor, das Mammakarzinom in 20 %, das Pankreaskarzinom und das Gallenblasenkarzinom jeweils in 6 %, der Ovarialtumor in 4 %, das Ösophaguskarzinom und andere Tumorentitäten jeweils in 2 % der Fälle. Die Lage der Lebermetastasen und die morphologische Veränderung der Läsionen in Größe, Volumen, nekrotischem Bereich, Diffusion und ADC-Wert wurden durch Magnetresonanztomo-graphie (MRT) bewertet. Die erste MRT-Kontrolle erfolgte vor der Behandlung mit thermischer Ablation. Das Follow-up beinhaltet MRT-Kontrollen 24 Stunden nach der Ablation und 4 Kontrolluntersuchungen im Abstand von 3 Monaten innerhalb von einem Jahr.
50 Läsionen mit einem mittleren Durchmesser von 1,63 cm (Range: 0,6 - 3,7 cm; MWA 1,72 cm, RFA 1,53 cm) wurden mit thermischer Ablation behandelt. Die lokale Rezidivrate innerhalb eines Jahres betrug für beide Versuchsarme 4 % (2/50), 0 % (0/26) in der MWA-Gruppe und 8,3 % (2/24) in der RFA-Gruppe. Die Rate für neu entstandene maligne Formationen, die sich an einem anderen Ort als die behandelte Läsion befanden, betrug in beiden Gruppen etwa 38 % (19/50), in der MWA-Gruppe 38,4 % und in der RFA-Gruppe 37,5 %. Die Sterblichkeitsrate für diese Studie betrug 14,0 %. 15,4 % der Patienten im Behandlungsarm der MWA-Gruppe und 12,5 % im RFA-Behandlungsarm verstarben im Rahmen dieser Studie. Es konnten keine größeren Komplikationen festgestellt werden.
Zusammenfassend zeigt die MIRA-Studie zur thermischen Ablation von Lebermetastasen keine signifikanten Unterschiede in der Mortalität, malignen Neubildungen oder Komplikationsraten zwischen MWA und RFA. Die Studie stellt fest, dass die thermische Behandlung mit MWA im Rahmen des 1-Jahres-Follow-ups größere Volumina im Ablationsgebiet und eine gering höhere Mortalität hervorruft, wobei man die höhere Pankreaskarzinomrate in diesem Studienzweig berücksichtigen sollte. Im RFA-Studienarm wurden hingegen 2 Lokalrezidive erfasst.
Vor dem Hintergrund der hier vorliegenden Ergebnisse und in Zusammenschau mit der aktuellen Literatur sind beide thermischen Verfahren mehrheitlich sicher durchführbare und effektive Methoden zur Behandlung von Patienten mit nicht operablen sekundären Lebermalignomen. Unterschiede zeigen sich in den physikalischen, technischen Voraussetzungen und damit in der Größe der erzeugten Ablations- und Nekrosezonen. Diese spielen vorrangig in der Behandlung von großen und gefäß- sowie gallengangsnahen Läsionen eine differenzierte Rolle. Die Größe und die Heterogenität des Patientenkollektivs in Bezug auf Tumorentität, die unterschiedlichen Vorbehandlungen sowie der Nachbeobachtungszeit-raum von nur 12 Monaten mindern die Aussagekraft der Studie. Um die vorliegenden Ergebnisse zu stützen, sollten weitere Untersuchungen mit einem größe-ren Patientenkollektiv folgen, beispielsweise in Form einer multizentrischen Studie, in welcher differenzierter bezüglich Tumorentität und Vorbehandlungen unterteilt wird.
Characteristics of critical incident reporting systems in primary care: an international survey
(2022)
Aim: The aim of the study was to support the development of future critical incident reporting systems (CIRS) in primary care by collecting information on existing systems. Our focus was on processes used to report and analyse incidents, as well as strategies used to overcome difficulties.
Methods: Based on literature from throughout the world, we identified existing CIRS in primary care. We developed a questionnaire and sent it to operators of a purposeful sample of 17 CIRS in primary care. We used cross-case analysis to compare the answers and pinpoint important similarities and differences in the CIRS in our sample.
Results: Ten CIRS operators filled out the questionnaire, and 9 systems met the inclusion criteria. The sample of CIRS came from 8 different countries and was rather heterogeneous. The reporting systems invited a broad range of professions to report, with some also including reports by patients. In most cases, reporting was voluntary and conducted via an online reporting form. Reports were analysed locally, centrally, or both. The various CIRS used interesting ideas to deal with barriers. Some, for example, used confidential reporting modes as a compromise between anonymity and the need for follow-up investigations, whereas others used smartphone applications and call centres to speed up the reporting process.
Conclusion: We found multiple CIRS that have operated in primary care for many years, have received a high number of reports and were largely developed in accordance with recommendations found in literature. Although primary care in Germany differs from other countries, these CIRS could serve as an inspiration for CIRS in German primary care.
Acute myeloid leukemia (AML) is a neoplastic disease of an early myeloid precursor cell in hematopoiesis. It leads to the accumulation of monoclonal cells in the bone marrow and the peripheral blood, showing a differentiation block and deregulated self-renewal. Frequently, the leukemic cells exhibit genetic aberrations with reciprocal chromosomal translocations. These translocations induce the formation of a fusion protein, that can lead to new cellular functions and a transformation into a leukemic cell. Common chromosomal translocation in AML are t(8;21) or t(15;17), which cause the formation of the fusion proteins AML1/ETO and PML/RARα and determine the leukemic phenotype of the AML.
The translocation t(6;9) leads to the formation of the fusion protein DEK/CAN and is of special interest, because of its association with mostly young patients and a very aggressive course of the disease. The fusion product induces leukemia in a small subset of hematopoietic stem cells, but its mechanism of leukemogenesis is greatly unknown.
The intention of this work was to characterize the DEK/CAN-induced AML on a molecular genetic level to gain a deeper understanding of the disease pathogenesis. Therefore, gene expression analysis with polymerase chain reaction (PCR) and microarray analysis was performed.
To detect DEK/CAN in different cell lines by PCR and real-time quantitative PCR (qPCR), specific primers and probes were designed, and a standardized workflow was established. Emphasis was placed on the optimization of RNA isolation, DNase treatment, cDNA synthesis with following PCR and qPCR, which enabled the detection of the fusion product DEK/CAN in the cell lines 32B, Phoenix and FKH-1. To quantify the fusion product DEK/CAN, the method of qPCR with absolute and relative quantification was used. Absolute quantification enabled the calculation of an exact copy number of the fusion transcript DEK/CAN with a detection limit of 50 copies/µl at a sensitivity of 10-6, which is of importance in determining the minimal residual disease (MRD) of patients with DEK/CAN-positive AML. MRD detection by qPCR is a highly sensitive diagnostic method to identify leukemic cells, even in low cell counts. This enables a thorough evaluation of the treatment response and allows an early detection of changes in the MRD level as part of the remission control.
Additionally, a microarray gene expression analysis was performed to identify alterations in relevant target genes and associated signaling pathways in DEK/CAN-positive cells.
Because of DEK/CAN’s potential to induce leukemia in a subset of hematopoietic stem cells, Sca+/Lin- cells of the bone marrow of C57Bl/6 mice were used and transfected with the gene products DEK/CAN and PML/RARα. Microarray analysis led to the identification of 16 different genes of interest, which demonstrated significant alterations of gene expression in DEK/CAN-positive cells. They were validated and quantified with TaqMan assay assisted qPCR. The elevated expression of the transcription factors TRIM25, HIF1α and ATF2, in DEK/CAN-positive cells, indicated an altered transcription factor activity and interaction with DNA in the nucleus. The localization of DEK/CAN in the nucleus emphasizes this assumption. Also, the upregulated expression of the nuclear export receptor XPO1 suggested changes in nuclear transport processes and impaired export activity in DEK/CAN-positive cells.
Furthermore, the results demonstrated changes of gene expression in genes that are involved in the JAK/STAT signaling pathway. PTPRC, the Protein Tyrosine Phosphatase Receptor Type C, functions as a direct inhibitor of JAKs (Janus Kinases) and STATs (Signal Transducers and Activators of Transcription) and their associated signaling pathway.
It was shown that the gene expression of PTPRC was significantly reduced in DEK/CAN-positive cells. This allowed the assumption, that the reduced expression of PTPRC led to a loss of inhibition and thus a consecutive hyperactivation of the JAK/STAT signaling pathway. This hypothesis was supported by an independent activation of PIM1, a target gene of STAT5 and the activation of LMO2, a direct target gene of JAK2. In addition, the transmembrane receptor CSF1R, which is directly involved in STAT activation, also showed an upregulation in gene expression.
The results of this work show an activation of the JAK/STAT signaling pathway in DEK/CAN-positive cells, which may be a key mechanism in DEK/CAN-induced leukemogenesis.
Considering treatment options in the future, the addition of targeted therapy, such as pan-JAK inhibitors, to the standard therapy, could be a chance to improve the overall survival rate and the prognosis of t(6;9)-positive AML.