TY - JOUR A1 - Lisowska, Justyna A1 - Rödel, Claudia Jasmin A1 - Manet, Sandra A1 - Miroshnikova, Yekaterina A. A1 - Boyault, Cyril A1 - Planus, Emmanuelle A1 - De Mets, Richard A1 - Lee, Hsiao-Hui A1 - Destaing, Olivier A1 - Mertani, Hichem A1 - Boulday, Gwenola A1 - Tournier-Lasserve, Elisabeth A1 - Balland, Martial A1 - Abdelilah-Seyfried, Salim A1 - Albiges-Rizo, Corinne A1 - Faurobert, Eva T1 - The CCM1-CCM2 complex controls complementary functions of ROCK1 and ROCK2 that are required for endothelial integrity JF - Journal of cell science N2 - Endothelial integrity relies on a mechanical crosstalk between intercellular and cell-matrix interactions. This crosstalk is compromised in hemorrhagic vascular lesions of patients carrying loss-of-function mutations in cerebral cavernous malformation (CCM) genes. RhoA/ROCK-dependent cytoskeletal remodeling is central to the disease, as it causes unbalanced cell adhesion towards increased cell-extracellular matrix adhesions and destabilized cell-cell junctions. This study reveals that CCM proteins directly orchestrate ROCK1 and ROCK2 complementary roles on the mechanics of the endothelium. CCM proteins act as a scaffold, promoting ROCK2 interactions with VE-cadherin and limiting ROCK1 kinase activity. Loss of CCM1 (also known as KRIT1) produces excessive ROCK1-dependent actin stress fibers and destabilizes intercellular junctions. Silencing of ROCK1 but not ROCK2 restores the adhesive and mechanical homeostasis of CCM1 and CCM2-depleted endothelial monolayers, and rescues the cardiovascular defects of ccm1 mutant zebrafish embryos. Conversely, knocking down Rock2 but not Rock1 in wild-type zebrafish embryos generates defects reminiscent of the ccm1 mutant phenotypes. Our study uncovers the role of the CCM1-CCM2 complex in controlling ROCK1 and ROCK2 to preserve endothelial integrity and drive heart morphogenesis. Moreover, it solely identifies the ROCK1 isoform as a potential therapeutic target for the CCM disease. KW - CCM KW - ROCK KW - Endothelial integrity KW - Mechanotransduction Y1 - 2018 U6 - https://doi.org/10.1242/jcs.216093 SN - 0021-9533 SN - 1477-9137 VL - 131 IS - 15 PB - Company biologists LTD CY - Cambridge ER - TY - JOUR A1 - Otten, Cecile A1 - Knox, Jessica A1 - Boulday, Gwenola A1 - Eymery, Mathias A1 - Haniszewski, Marta A1 - Neuenschwander, Martin A1 - Radetzki, Silke A1 - Vogt, Ingo A1 - Haehn, Kristina A1 - De Luca, Coralie A1 - Cardoso, Cecile A1 - Hamad, Sabri A1 - Igual Gil, Carla A1 - Roy, Peter A1 - Albiges-Rizo, Corinne A1 - Faurobert, Eva A1 - von Kries, Jens P. A1 - Campillos, Monica A1 - Tournier-Lasserve, Elisabeth A1 - Derry, William Brent A1 - Abdelilah-Seyfried, Salim T1 - Systematic pharmacological screens uncover novel pathways involved in cerebral cavernous malformations JF - EMBO molecular medicine N2 - Cerebral cavernous malformations (CCMs) are vascular lesions in the central nervous system causing strokes and seizures which currently can only be treated through neurosurgery. The disease arises through changes in the regulatory networks of endothelial cells that must be comprehensively understood to develop alternative, non-invasive pharmacological therapies. Here, we present the results of several unbiased small-molecule suppression screens in which we applied a total of 5,268 unique substances to CCM mutant worm, zebrafish, mouse, or human endothelial cells. We used a systems biology-based target prediction tool to integrate the results with the whole-transcriptome profile of zebrafish CCM2 mutants, revealing signaling pathways relevant to the disease and potential targets for small-molecule-based therapies. We found indirubin-3-monoxime to alleviate the lesion burden in murine preclinical models of CCM2 and CCM3 and suppress the loss-of-CCM phenotypes in human endothelial cells. Our multi-organism-based approach reveals new components of the CCM regulatory network and foreshadows novel small-molecule-based therapeutic applications for suppressing this devastating disease in patients. KW - angiogenesis KW - CCM KW - ERK5 KW - indirubin-3-monoxime KW - KLF2 Y1 - 2018 U6 - https://doi.org/10.15252/emmm.201809155 SN - 1757-4676 SN - 1757-4684 VL - 10 IS - 10 PB - Wiley CY - Hoboken ER - TY - JOUR A1 - Bornhorst, Dorothee A1 - Abdelilah-Seyfried, Salim T1 - Strong as a Hippo’s Heart: Biomechanical Hippo Signaling During Zebrafish Cardiac Development JF - Frontiers in Cell and Developmental Biology N2 - The heart is comprised of multiple tissues that contribute to its physiological functions. During development, the growth of myocardium and endocardium is coupled and morphogenetic processes within these separate tissue layers are integrated. Here, we discuss the roles of mechanosensitive Hippo signaling in growth and morphogenesis of the zebrafish heart. Hippo signaling is involved in defining numbers of cardiac progenitor cells derived from the secondary heart field, in restricting the growth of the epicardium, and in guiding trabeculation and outflow tract formation. Recent work also shows that myocardial chamber dimensions serve as a blueprint for Hippo signaling-dependent growth of the endocardium. Evidently, Hippo pathway components act at the crossroads of various signaling pathways involved in embryonic zebrafish heart development. Elucidating how biomechanical Hippo signaling guides heart morphogenesis has direct implications for our understanding of cardiac physiology and pathophysiology. KW - Hippo signaling KW - Yap1/Wwtr1 (Taz) KW - cardiac development KW - mechanobiology KW - endocardium KW - myocardium KW - zebrafish KW - intra-organ-communication Y1 - 2021 U6 - https://doi.org/10.3389/fcell.2021.731101 SN - 2296-634X VL - 9 SP - 1 EP - 10 PB - Frontiers Media CY - Lausanne, Schweiz ER - TY - JOUR A1 - Münch, Juliane A1 - Abdelilah-Seyfried, Salim T1 - Sensing and responding of cardiomyocytes to changes of tissue stiffness in the diseased heart JF - Frontiers in cell developmental biology N2 - Cardiomyocytes are permanently exposed to mechanical stimulation due to cardiac contractility. Passive myocardial stiffness is a crucial factor, which defines the physiological ventricular compliance and volume of diastolic filling with blood. Heart diseases often present with increased myocardial stiffness, for instance when fibrotic changes modify the composition of the cardiac extracellular matrix (ECM). Consequently, the ventricle loses its compliance, and the diastolic blood volume is reduced. Recent advances in the field of cardiac mechanobiology revealed that disease-related environmental stiffness changes cause severe alterations in cardiomyocyte cellular behavior and function. Here, we review the molecular mechanotransduction pathways that enable cardiomyocytes to sense stiffness changes and translate those into an altered gene expression. We will also summarize current knowledge about when myocardial stiffness increases in the diseased heart. Sophisticated in vitro studies revealed functional changes, when cardiomyocytes faced a stiffer matrix. Finally, we will highlight recent studies that described modulations of cardiac stiffness and thus myocardial performance in vivo. Mechanobiology research is just at the cusp of systematic investigations related to mechanical changes in the diseased heart but what is known already makes way for new therapeutic approaches in regenerative biology. KW - mechanobiology KW - tissue stiffness KW - cardiomyocyte KW - heart regeneration KW - titin KW - collagen KW - agrin KW - extracellular matrix Y1 - 2020 U6 - https://doi.org/10.3389/fcell.2021.642840 SN - 2296-634X VL - 9 PB - Frontiers Media CY - Lausanne ER - TY - JOUR A1 - Renz, Marc A1 - Otten, Cecile A1 - Faurobert, Eva A1 - Rudolph, Franziska A1 - Zhu, Yuan A1 - Boulday, Gwenola A1 - Duchene, Johan A1 - Mickoleit, Michaela A1 - Dietrich, Ann-Christin A1 - Ramspacher, Caroline A1 - Steed, Emily A1 - Manet-Dupe, Sandra A1 - Benz, Alexander A1 - Hassel, David A1 - Vermot, Julien A1 - Huisken, Jan A1 - Tournier-Lasserve, Elisabeth A1 - Felbor, Ute A1 - Sure, Ulrich A1 - Albiges-Rizo, Corinne A1 - Abdelilah-Seyfried, Salim T1 - Regulation of beta 1 Integrin-Klf2-Mediated angiogenesis by CCM proteins JF - Developmental cell N2 - Mechanotransduction pathways are activated in response to biophysical stimuli during the development or homeostasis of organs and tissues. In zebrafish, the blood-flow-sensitive transcription factor Klf2a promotes VEGF-dependent angiogenesis. However, the means by which the Klf2a mechanotransduction pathway is regulated to prevent continuous angiogenesis remain unknown. Here we report that the upregulation of klf2 mRNA causes enhanced egfl7 expression and angiogenesis signaling, which underlies cardiovascular defects associated with the loss of cerebral cavernous malformation (CCM) proteins in the zebrafish embryo. Using CCM-protein-depleted human umbilical vein endothelial cells, we show that the misexpression of KLF2 mRNA requires the extracellular matrix-binding receptor beta 1 integrin and occurs in the absence of blood flow. Downregulation of beta 1 integrin rescues ccm mutant cardiovascular malformations in zebrafish. Our work reveals a beta 1 integrin-Klf2-Egfl7-signaling pathway that is tightly regulated by CCM proteins. This regulation prevents angiogenic overgrowth and ensures the quiescence of endothelial cells. Y1 - 2015 U6 - https://doi.org/10.1016/j.devcel.2014.12.016 SN - 1534-5807 SN - 1878-1551 VL - 32 IS - 2 SP - 181 EP - 190 PB - Cell Press CY - Cambridge ER - TY - JOUR A1 - Merks, Anne Margarete A1 - Swinarski, Marie A1 - Meyer, Alexander Matthias A1 - Müller, Nicola Victoria A1 - Özcan, Ismail A1 - Donat, Stefan A1 - Burger, Alexa A1 - Gilbert, Stephen A1 - Mosimann, Christian A1 - Abdelilah-Seyfried, Salim A1 - Panakova, Daniela T1 - Planar cell polarity signalling coordinates heart tube remodelling through tissue-scale polarisation of actomyosin activity JF - Nature Communications N2 - Development of a multiple-chambered heart from the linear heart tube is inherently linked to cardiac looping. Although many molecular factors regulating the process of cardiac chamber ballooning have been identified, the cellular mechanisms underlying the chamber formation remain unclear. Here, we demonstrate that cardiac chambers remodel by cell neighbour exchange of cardiomyocytes guided by the planar cell polarity (PCP) pathway triggered by two non-canonical Wnt ligands, Wnt5b and Wnt11. We find that PCP signalling coordinates the localisation of actomyosin activity, and thus the efficiency of cell neighbour exchange. On a tissue-scale, PCP signalling planar-polarises tissue tension by restricting the actomyosin contractility to the apical membranes of outflow tract cells. The tissue-scale polarisation of actomyosin contractility is required for cardiac looping that occurs concurrently with chamber ballooning. Taken together, our data reveal that instructive PCP signals couple cardiac chamber expansion with cardiac looping through the organ-scale polarisation of actomyosin-based tissue tension. Y1 - 2018 U6 - https://doi.org/10.1038/s41467-018-04566-1 SN - 2041-1723 VL - 9 PB - Nature Publ. Group CY - London ER - TY - JOUR A1 - Cui, Huanhuan A1 - Schlesinger, Jenny A1 - Schoenhals, Sophia A1 - Toenjes, Martje A1 - Dunkel, Ilona A1 - Meierhofer, David A1 - Cano, Elena A1 - Schulz, Kerstin A1 - Berger, Michael F. A1 - Haack, Timm A1 - Abdelilah-Seyfried, Salim A1 - Bulyk, Martha L. A1 - Sauer, Sascha A1 - Sperling, Silke R. T1 - Phosphorylation of the chromatin remodeling factor DPF3a induces cardiac hypertrophy through releasing HEY repressors from DNA JF - Nucleic acids research N2 - DPF3 (BAF45c) is a member of the BAF chromatin remodeling complex. Two isoforms have been described, namely DPF3a and DPF3b. The latter binds to acetylated and methylated lysine residues of histones. Here, we elaborate on the role of DPF3a and describe a novel pathway of cardiac gene transcription leading to pathological cardiac hypertrophy. Upon hypertrophic stimuli, casein kinase 2 phosphorylates DPF3a at serine 348. This initiates the interaction of DPF3a with the transcriptional repressors HEY, followed by the release of HEY from the DNA. Moreover, BRG1 is bound by DPF3a, and is thus recruited to HEY genomic targets upon interaction of the two components. Consequently, the transcription of downstream targets such as NPPA and GATA4 is initiated and pathological cardiac hypertrophy is established. In human, DPF3a is significantly up-regulated in hypertrophic hearts of patients with hypertrophic cardiomyopathy or aortic stenosis. Taken together, we show that activation of DPF3a upon hypertrophic stimuli switches cardiac fetal gene expression from being silenced by HEY to being activated by BRG1. Thus, we present a novel pathway for pathological cardiac hypertrophy, whose inhibition is a long-term therapeutic goal for the treatment of the course of heart failure. Y1 - 2016 U6 - https://doi.org/10.1093/nar/gkv1244 SN - 0305-1048 SN - 1362-4962 VL - 44 SP - 2538 EP - 2553 PB - Oxford Univ. Press CY - Oxford ER - TY - JOUR A1 - Lombardo, Verónica A. A1 - Heise, Melina A1 - Moghtadaei, Motahareh A1 - Bornhorst, Dorothee A1 - Männer, Jörg A1 - Abdelilah-Seyfried, Salim T1 - Morphogenetic control of zebrafish cardiac looping by Bmp signaling JF - Development : Company of Biologists N2 - Cardiac looping is an essential and highly conserved morphogenetic process that places the different regions of the developing vertebrate heart tube into proximity of their final topographical positions. High-resolution 4D live imaging of mosaically labelled cardiomyocytes reveals distinct cardiomyocyte behaviors that contribute to the deformation of the entire heart tube. Cardiomyocytes acquire a conical cell shape, which is most pronounced at the superior wall of the atrioventricular canal and contributes to S-shaped bending. Torsional deformation close to the outflow tract contributes to a torque-like winding of the entire heart tube between its two poles. Anisotropic growth of cardiomyocytes based on their positions reinforces S-shaping of the heart. During cardiac looping, bone morphogenetic protein pathway signaling is strongest at the future superior wall of the atrioventricular canal. Upon pharmacological or genetic inhibition of bone morphogenetic protein signaling, myocardial cells at the superior wall of the atrioventricular canal maintain cuboidal cell shapes and S-shaped bending is impaired. This description of cellular rearrangements and cardiac looping regulation may also be relevant for understanding the etiology of human congenital heart defects. KW - BMP KW - Wnt KW - Cardiac looping KW - Hemodynamics KW - Zebrafish Y1 - 2019 U6 - https://doi.org/10.1242/dev.180091 SN - 0950-1991 SN - 1477-9129 VL - 146 IS - 22 PB - The Company of Biologists Ltd CY - Cambridge ER - TY - JOUR A1 - Lombardo, Veronica A. A1 - Otten, Cecile A1 - Abdelilah-Seyfried, Salim T1 - Large-scale Zebrafish Embryonic Heart Dissection for Transcriptional Analysis JF - Journal of visualized experiments N2 - The zebrafish embryonic heart is composed of only a few hundred cells, representing only a small fraction of the entire embryo. Therefore, to prevent the cardiac transcriptome from being masked by the global embryonic transcriptome, it is necessary to collect sufficient numbers of hearts for further analyses. Furthermore, as zebrafish cardiac development proceeds rapidly, heart collection and RNA extraction methods need to be quick in order to ensure homogeneity of the samples. Here, we present a rapid manual dissection protocol for collecting functional/beating hearts from zebrafish embryos. This is an essential prerequisite for subsequent cardiac-specific RNA extraction to determine cardiac-specific gene expression levels by transcriptome analyses, such as quantitative real-time polymerase chain reaction (RT-qPCR). The method is based on differential adhesive properties of the zebrafish embryonic heart compared with other tissues; this allows for the rapid physical separation of cardiac from extracardiac tissue by a combination of fluidic shear force disruption, stepwise filtration and manual collection of transgenic fluorescently labeled hearts. KW - Developmental Biology KW - Issue 95 KW - zebrafish KW - embryo KW - heart KW - dissection KW - RNA KW - RT-qPCR Y1 - 2015 U6 - https://doi.org/10.3791/52087 SN - 1940-087X IS - 95 PB - JoVE CY - Cambridge ER - TY - JOUR A1 - Donat, Stefan A1 - Lourenco, Marta Sofia Rocha A1 - Paolini, Alessio A1 - Otten, Cecile A1 - Renz, Marc A1 - Abdelilah-Seyfried, Salim T1 - Heg1 and Ccm1/2 proteins control endocardial mechanosensitivity during zebrafish valvulogenesis JF - eLife N2 - Endothelial cells respond to different levels of fluid shear stress through adaptations of their mechanosensitivity. Currently, we lack a good understanding of how this contributes to sculpting of the cardiovascular system. Cerebral cavernous malformation (CCM) is an inherited vascular disease that occurs when a second somatic mutation causes a loss of CCM1/KRIT1, CCM2, or CCM3 proteins. Here, we demonstrate that zebrafish Krit1 regulates the formation of cardiac valves. Expression of heg1, which encodes a binding partner of Krit1, is positively regulated by blood-flow. In turn, Heg1 stabilizes levels of Krit1 protein, and both Heg1 and Krit1 dampen expression levels of klf2a, a major mechanosensitive gene. Conversely, loss of Krit1 results in increased expression of klf2a and notch1b throughout the endocardium and prevents cardiac valve leaflet formation. Hence, the correct balance of blood-flow-dependent induction and Krit1 protein mediated repression of klf2a and notch1b ultimately shapes cardiac valve leaflet morphology. Y1 - 2018 U6 - https://doi.org/10.7554/eLife.28939 SN - 2050-084X VL - 7 PB - eLife Sciences Publications CY - Cambridge ER -