{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:33:52Z","timestamp":1787027632680,"version":"build-2736575974"},"reference-count":35,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2018,11,19]],"date-time":"2018-11-19T00:00:00Z","timestamp":1542585600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Patients with inherited dilated cardiomyopathy (DCM) often suffer from severe heart failure based on impaired cardiac contractility leading to increased morbidity and mortality. Integrin-linked kinase (ILK) as a part of the cardiac mechanical stretch sensor was found to be an essential genetic regulator of cardiac contractility. Integrin-linked kinase localizes to z-disks and costameres in vertebrate hearts and regulates the activity of the signaling molecule protein kinase B (PKB\/Akt) by controlling its phosphorylation. Despite identification of several potential drug targets in the ILK signaling pathway, pharmacological treatment strategies to restore contractile function in ILK-dependent cardiomyopathies have not been established yet. In recent years, the zebrafish has emerged as a valuable experimental system to model human cardiomyopathies as well as a powerful tool for the straightforward high-throughput in vivo small compound screening of therapeutically active substances. Using the ILK deficient zebrafish heart failure mutant main squeeze (msq), which shows reduced PKB phosphorylation and thereby impaired cardiac contractile force, we identified here, in an automated small compound screen, the protein phosphatase inhibitors calyculin A and okadaic acid significantly restoring myocardial contractile function by reconstituting PKB phosphorylation in msq ILK-deficient zebrafish embryos.<\/jats:p>","DOI":"10.3390\/biom8040153","type":"journal-article","created":{"date-parts":[[2018,11,21]],"date-time":"2018-11-21T11:23:27Z","timestamp":1542799407000},"page":"153","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Therapeutic Chemical Screen Identifies Phosphatase Inhibitors to Reconstitute PKB Phosphorylation and Cardiac Contractility in ILK-Deficient Zebrafish"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5824-8098","authenticated-orcid":false,"given":"Alexander","family":"Pott","sequence":"first","affiliation":[{"name":"Department of Internal Medicine II, Ulm University, Albert-Einstein-Allee 23, D-89081 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maryam","family":"Shahid","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine II, Ulm University, Albert-Einstein-Allee 23, D-89081 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Doreen","family":"K\u00f6hler","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine III, University of Heidelberg, D-69120 Heidelberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian","family":"Pylatiuk","sequence":"additional","affiliation":[{"name":"Institute of Applied Computer Science, Karlsruhe Institute of Technology, D-76344 Eggenstein-Leopoldshafen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karolina","family":"Weinmann","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine II, Ulm University, Albert-Einstein-Allee 23, D-89081 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steffen","family":"Just","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine II, Ulm University, Albert-Einstein-Allee 23, D-89081 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wolfgang","family":"Rottbauer","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine II, Ulm University, Albert-Einstein-Allee 23, D-89081 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,11,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1161\/CIRCRESAHA.117.309711","article-title":"Classification, Epidemiology, and Global Burden of Cardiomyopathies","volume":"121","author":"McKenna","year":"2017","journal-title":"Circ. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1007\/s00108-011-2988-z","article-title":"Clinical and genetic aspects of hypertrophic and dilated cardiomyopathy","volume":"53","author":"Meder","year":"2012","journal-title":"Der Internist"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.hfc.2017.12.007","article-title":"Epidemiology and Clinical Aspects of Genetic Cardiomyopathies","volume":"14","author":"Masarone","year":"2018","journal-title":"Heart Fail. Clin."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1093\/eurheartj\/ehm585","article-title":"Revisiting definition and classification of cardiomyopathies in the era of molecular medicine","volume":"29","author":"Thiene","year":"2008","journal-title":"Eur. Heart J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1093\/eurheartj\/ehu301","article-title":"Atlas of the clinical genetics of human dilated cardiomyopathy","volume":"36","author":"Haas","year":"2015","journal-title":"Eur. Heart J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"750","DOI":"10.1007\/s00109-003-0488-x","article-title":"Cardiac mechanotransduction and implications for heart disease","volume":"81","author":"Knoll","year":"2003","journal-title":"J. Mol. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1161\/CIRCULATIONAHA.107.689984","article-title":"Laminin-\u03b14 and integrin-linked kinase mutations cause human cardiomyopathy via simultaneous defects in cardiomyocytes and endothelial cells","volume":"116","author":"Knoll","year":"2007","journal-title":"Circulation"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"H1313","DOI":"10.1152\/ajpheart.00816.2005","article-title":"Mechanical stress-strain sensors embedded in cardiac cytoskeleton: Z disk, titin, and associated structures","volume":"290","author":"Hoshijima","year":"2006","journal-title":"Am. J. Physiol. Heart Circ. Physiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2355","DOI":"10.1101\/gad.1458906","article-title":"Targeted ablation of ILK from the murine heart results in dilated cardiomyopathy and spontaneous heart failure","volume":"20","author":"White","year":"2006","journal-title":"Genes Dev."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2361","DOI":"10.1101\/gad.1448306","article-title":"Integrin-linked kinase, a novel component of the cardiac mechanical stretch sensor, controls contractility in the zebrafish heart","volume":"20","author":"Bendig","year":"2006","journal-title":"Genes Dev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1083\/jcb.153.3.585","article-title":"A new focal adhesion protein that interacts with integrin-linked kinase and regulates cell adhesion and spreading","volume":"153","author":"Tu","year":"2001","journal-title":"J. Cell Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1016\/j.ceb.2012.06.003","article-title":"ILK: A pseudokinase in the center stage of cell-matrix adhesion and signaling","volume":"24","author":"Qin","year":"2012","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1038\/nrm1789","article-title":"ILK, PINCH and parvin: The tIPP of integrin signalling","volume":"7","author":"Legate","year":"2006","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"7294","DOI":"10.1093\/emboj\/17.24.7294","article-title":"Inactivation and dephosphorylation of protein kinase B\u03b1 (PKB\u03b1) promoted by hyperosmotic stress","volume":"17","author":"Meier","year":"1998","journal-title":"EMBO J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3424","DOI":"10.1128\/MCB.05269-11","article-title":"PINCH proteins regulate cardiac contractility by modulating integrin-linked kinase-protein kinase B signaling","volume":"31","author":"Meder","year":"2011","journal-title":"Mol. Cell. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/S0167-4781(02)00247-6","article-title":"Promoter characterization and genomic organization of the gene encoding integrin-linked kinase 1","volume":"1575","author":"Melchior","year":"2002","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"926","DOI":"10.1101\/gad.255603","article-title":"Integrin-linked kinase (ILK) is required for polarizing the epiblast, cell adhesion, and controlling actin accumulation","volume":"17","author":"Sakai","year":"2003","journal-title":"Genes Dev."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1038\/379091a0","article-title":"Regulation of cell adhesion and anchorage-dependent growth by a new \u03b2 1-integrin-linked protein kinase","volume":"379","author":"Hannigan","year":"1996","journal-title":"Nature"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1016\/S0960-9822(02)00810-2","article-title":"C. elegans PAT-4\/ILK functions as an adaptor protein within integrin adhesion complexes","volume":"12","author":"Mackinnon","year":"2002","journal-title":"Curr. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"21886","DOI":"10.1074\/jbc.M111.240093","article-title":"Biochemical, proteomic, structural, and thermodynamic characterizations of integrin-linked kinase (ILK): Cross-validation of the pseudokinase","volume":"286","author":"Fukuda","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1177\/2211068212464223","article-title":"High-throughput screening of zebrafish embryos using automated heart detection and imaging","volume":"17","author":"Spomer","year":"2012","journal-title":"J. Lab. Autom."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1089\/zeb.2014.1002","article-title":"Automatic zebrafish heartbeat detection and analysis for zebrafish embryos","volume":"11","author":"Pylatiuk","year":"2014","journal-title":"Zebrafish"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1320","DOI":"10.2174\/0929867321666131227130218","article-title":"Functional genomics in zebrafish as a tool to identify novel antiarrhythmic targets","volume":"21","author":"Pott","year":"2014","journal-title":"Curr. Med. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1624","DOI":"10.1101\/gad.1319405","article-title":"VEGF-PLC\u03b31 pathway controls cardiac contractility in the embryonic heart","volume":"19","author":"Rottbauer","year":"2005","journal-title":"Genes Dev."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"D646","DOI":"10.2741\/Dawson","article-title":"Molecular mechanisms underlying inhibition of protein phosphatases by marine toxins","volume":"4","author":"Dawson","year":"1999","journal-title":"Front. Biosci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"11369","DOI":"10.1074\/jbc.M109.028142","article-title":"Immunosuppressive cyclosporin A activates AKT in keratinocytes through PTEN suppression: Implications in skin carcinogenesis","volume":"285","author":"Han","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1016\/0041-0101(94)90086-8","article-title":"Characterization of natural toxins with inhibitory activity against serine\/threonine protein phosphatases","volume":"32","author":"Honkanen","year":"1994","journal-title":"Toxicon"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1016\/j.diff.2012.05.005","article-title":"Zebrafish models in cardiac development and congenital heart birth defects","volume":"84","author":"Tu","year":"2012","journal-title":"Differentiation"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"650","DOI":"10.1161\/CIRCRESAHA.108.186676","article-title":"A single serine in the carboxyl terminus of cardiac essential myosin light chain-1 controls cardiomyocyte contractility in vivo","volume":"104","author":"Meder","year":"2009","journal-title":"Circ. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"871","DOI":"10.1016\/0006-291X(89)92189-X","article-title":"Calyculin A and okadaic acid: Inhibitors of protein phosphatase activity","volume":"159","author":"Ishihara","year":"1989","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1042\/BST20160154","article-title":"Biogenesis and activity regulation of protein phosphatase 1","volume":"45","author":"Verbinnen","year":"2017","journal-title":"Biochem. Soc. Trans."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Baskaran, R., and Velmurugan, B.K. (2018). Protein phosphatase 2A as therapeutic targets in various disease models. Life Sci.","DOI":"10.1016\/j.lfs.2018.08.063"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1161\/01.RES.84.6.735","article-title":"Cyclosporine attenuates pressure-overload hypertrophy in mice while enhancing susceptibility to decompensation and heart failure","volume":"84","author":"Meguro","year":"1999","journal-title":"Circ. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1002\/bdrc.20212","article-title":"The developing zebrafish (Danio rerio): A vertebrate model for high-throughput screening of chemical libraries","volume":"93","author":"Lessman","year":"2011","journal-title":"Birth Defects Res. C Embryo Today"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1016\/j.addr.2015.05.010","article-title":"Human engineered heart tissue as a model system for drug testing","volume":"96","author":"Eder","year":"2016","journal-title":"Adv. Drug Deliv. Rev."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/8\/4\/153\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:30:43Z","timestamp":1760196643000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/8\/4\/153"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,11,19]]},"references-count":35,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2018,12]]}},"alternative-id":["biom8040153"],"URL":"https:\/\/doi.org\/10.3390\/biom8040153","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,11,19]]}}}