{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T04:37:45Z","timestamp":1778560665133,"version":"3.51.4"},"reference-count":42,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2022,12,10]],"date-time":"2022-12-10T00:00:00Z","timestamp":1670630400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["HL-121500"],"award-info":[{"award-number":["HL-121500"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Diaphragm weakness frequently develops in mechanically ventilated critically ill patients and is associated with increased morbidity, including ventilator weaning failure, mortality, and health care costs. The mechanisms underlying diaphragm weakness are incompletely understood but may include the elastic properties of titin, a giant protein whose layout in the muscle\u2019s sarcomeres makes it an ideal candidate to sense ventilation-induced diaphragm unloading, resulting in downstream signaling through titin-binding proteins. In the current study, we investigated whether modulating titin stiffness affects the development of diaphragm weakness during mechanical ventilation. To this end, we ventilated genetically engineered mice with reduced titin stiffness (Rbm20\u0394RRM), and robust (Ttn\u0394IAjxn) or severely (Ttn\u0394112\u2013158) increased titin stiffness for 8 h, and assessed diaphragm contractility and protein expression of titin-binding proteins. Mechanical ventilation reduced the maximum active tension of the diaphragm in WT, Ttn\u0394IAjxn and Ttn\u0394112\u2013158 mice. However, in Rbm20\u0394RRM mice maximum active tension was preserved after ventilation. Analyses of titin binding proteins suggest that muscle ankyrin repeat proteins (MARPs) 1 and 2 may play a role in the adaptation of the diaphragm to mechanical ventilation, and the preservation of diaphragm contractility in Rbm20\u0394RRM mice. Thus, Rbm20\u0394RRM mice, expressing titin isoforms with lower stiffness, are protected from mechanical ventilation-induced diaphragm weakness, suggesting that titin elasticity may modulate the diaphragm\u2019s response to unloading during mechanical ventilation.<\/jats:p>","DOI":"10.3390\/ijms232415689","type":"journal-article","created":{"date-parts":[[2022,12,12]],"date-time":"2022-12-12T01:42:27Z","timestamp":1670809347000},"page":"15689","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Rbm20\u0394RRM Mice, Expressing a Titin Isoform with Lower Stiffness, Are Protected from Mechanical Ventilation-Induced Diaphragm Weakness"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0920-6139","authenticated-orcid":false,"given":"Marloes","family":"van den Berg","sequence":"first","affiliation":[{"name":"Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ 85724, USA"},{"name":"Department of Physiology, Amsterdam UMC, 1082 HZ Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6846-2154","authenticated-orcid":false,"given":"Eva L.","family":"Peters","sequence":"additional","affiliation":[{"name":"Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ 85724, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robbert J.","family":"van der Pijl","sequence":"additional","affiliation":[{"name":"Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ 85724, USA"},{"name":"Department of Physiology, Amsterdam UMC, 1082 HZ Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shengyi","family":"Shen","sequence":"additional","affiliation":[{"name":"Department of Physiology, Amsterdam UMC, 1082 HZ Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Leo M. A.","family":"Heunks","sequence":"additional","affiliation":[{"name":"Intensive Care Medicine, Erasmus MC, 3015 GD Rotterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henk L.","family":"Granzier","sequence":"additional","affiliation":[{"name":"Department of Physiology, Amsterdam UMC, 1082 HZ Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Coen A. C.","family":"Ottenheijm","sequence":"additional","affiliation":[{"name":"Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ 85724, USA"},{"name":"Department of Physiology, Amsterdam UMC, 1082 HZ Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1164\/rccm.201209-1668OC","article-title":"Diaphragm Dysfunction on Admission to the Intensive Care Unit: Prevalence, Risk Factors, and Prognostic Impact\u2014A Prospective Study","volume":"188","author":"Demoule","year":"2013","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1007\/s00134-015-4125-2","article-title":"Diaphragmatic Dysfunction in Patients with ICU-Acquired Weakness and Its Impact on Extubation Failure","volume":"42","author":"Jung","year":"2016","journal-title":"Intensive Care Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"R120","DOI":"10.1186\/cc12792","article-title":"Diaphragm Weakness in Mechanically Ventilated Critically Ill Patients","volume":"17","author":"Supinski","year":"2013","journal-title":"Crit. Care"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1033","DOI":"10.1183\/09031936.00010206","article-title":"Weaning from Mechanical Ventilation","volume":"29","author":"Boles","year":"2007","journal-title":"Eur. Respir. J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2253","DOI":"10.1001\/jama.2010.761","article-title":"Long-Term Acute Care Hospital Utilization after Critical Illness","volume":"303","author":"Kahn","year":"2010","journal-title":"JAMA\u2014J. Am. Med. Assoc."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1164\/rccm.201602-0367OC","article-title":"Coexistence and Impact of Limb Muscle and Diaphragm Weakness at Time of Liberation from Mechanical Ventilation in Medical Intensive Care Unit Patients","volume":"195","author":"Dres","year":"2017","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1080","DOI":"10.1164\/rccm.201503-0620OC","article-title":"Evolution of Diaphragm Thickness during Mechanical Ventilation: Impact of Inspiratory Effort","volume":"192","author":"Goligher","year":"2015","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"P211","DOI":"10.1186\/cc8443","article-title":"Increased Duration of Mechanical Ventilation Is Associated with Decreased Diaphragmatic Force","volume":"14","author":"Hermans","year":"2010","journal-title":"Crit. Care"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1097\/ALN.0b013e318261e7f8","article-title":"Rapid Onset of Specific Diaphragm Weakness in a Healthy Murine Model of Ventilator-Induced Diaphragmatic Dysfunction","volume":"117","author":"Mrozek","year":"2012","journal-title":"Anesthesiology"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1325","DOI":"10.1097\/00003246-200107000-00005","article-title":"Measurement of Twitch Transdiaphragmatic, Esophageal, and Endotracheal Tube Pressure with Bilateral Anterolateral Magnetic Phrenic Nerve Stimulation in Patients in the Intensive Care Unit","volume":"29","author":"Watson","year":"2001","journal-title":"Crit. Care Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1544","DOI":"10.1164\/rccm.201703-0501OC","article-title":"Diaphragm Atrophy and Weakness in the Absence of Mitochondrial Dysfunction in the Critically Ill","volume":"196","author":"Hooijman","year":"2017","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"L460","DOI":"10.1152\/ajplung.00076.2014","article-title":"Unaffected Contractility of Diaphragm Muscle Fibers in Humans on Mechanical Ventilation","volume":"307","author":"Hooijman","year":"2014","journal-title":"Am. J. Physiol.\u2014Lung Cell. Mol. Physiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1126","DOI":"10.1164\/rccm.201412-2214OC","article-title":"Diaphragm Muscle Fiber Weakness and Ubiquitin-Proteasome Activation in Critically Ill Patients","volume":"191","author":"Hooijman","year":"2015","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1164\/rccm.201004-0670OC","article-title":"Rapidly Progressive Diaphragmatic Weakness and Injury during Mechanical Ventilation in Humans","volume":"183","author":"Jaber","year":"2012","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1136\/thoraxjnl-2015-207559","article-title":"Prolonged Controlled Mechanical Ventilation in Humans Triggers Myofibrillar Contractile Dysfunction and Myofilament Protein Loss in the Diaphragm","volume":"71","author":"Hussain","year":"2016","journal-title":"Thorax"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1327","DOI":"10.1056\/NEJMoa070447","article-title":"Rapid Disuse Atrophy of Diaphragm Fibers in Mechanically Ventilated Humans","volume":"358","author":"Levine","year":"2008","journal-title":"N. Engl. J. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1002\/mus.23748","article-title":"Diaphragm Muscle Atrophy in the Mouse after Long-Term Mechanical Ventilation","volume":"48","author":"Tang","year":"2013","journal-title":"Muscle Nerve"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"702","DOI":"10.1007\/s00134-012-2504-5","article-title":"Titin and Diaphragm Dysfunction in Mechanically Ventilated Rats","volume":"38","author":"Hees","year":"2012","journal-title":"Intensive Care Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1164\/rccm.201709-1917OC","article-title":"Positive End-Expiratory Pressure Ventilation Induces Longitudinal Atrophy in Diaphragm Fibers","volume":"198","author":"Lindqvist","year":"2018","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1851","DOI":"10.1152\/japplphysiol.00881.2001","article-title":"Mechanical Ventilation Results in Progressive Contractile Dysfunction in the Diaphragm","volume":"92","author":"Powers","year":"2002","journal-title":"J. Appl. Physiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1186\/s13054-019-2409-6","article-title":"Mechanical Ventilation Causes Diaphragm Dysfunction in Newborn Lambs","volume":"23","author":"Liang","year":"2019","journal-title":"Crit. Care"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"C167","DOI":"10.1152\/ajpcell.00509.2018","article-title":"Diaphragm Contractile Weakness Due to Reduced Mechanical Loading: Role of Titin","volume":"317","author":"Granzier","year":"2019","journal-title":"Am. J. Physiol.\u2014Cell Physiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1126\/science.270.5234.293","article-title":"Titins: Giant Proteins in Charge of Muscle Ultrastructure and Elasticity","volume":"270","author":"Labeit","year":"1995","journal-title":"Science"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"740","DOI":"10.1002\/mus.20886","article-title":"Structure-Function Relations of the Giant Elastic Protein Titin in Striated and Smooth Muscle Cells","volume":"36","author":"Granzier","year":"2007","journal-title":"Muscle Nerve"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2217","DOI":"10.1083\/jcb.105.5.2217","article-title":"The Positional Stability of Thick Filaments in Activated Skeletal Muscle Depends on Sarcomere Length: Evidence for the Role of Titin Filaments","volume":"105","author":"Horrowits","year":"1987","journal-title":"J. Cell Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e202112925","DOI":"10.1085\/jgp.202112925","article-title":"Muscle Ankyrin Repeat Protein 1 (MARP1) Locks Titin to the Sarcomeric Thin Filament and Is a Passive Force Regulator","volume":"153","author":"Shen","year":"2021","journal-title":"J. Gen. Physiol."},{"key":"ref_27","first-page":"637","article-title":"Sense and Stretchability: The Role of Titin and Titin-Associated Proteins in Myocardial Stress-Sensing and Mechanical Dysfunction","volume":"77","author":"Linke","year":"2008","journal-title":"Cardiovasc. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1152\/physiol.00036.2019","article-title":"Titin: A Tunable Spring in Active Muscle","volume":"35","author":"Nishikawa","year":"2020","journal-title":"Physiology"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1499","DOI":"10.1016\/j.bpj.2011.01.059","article-title":"Thick-Filament Strain and Interfilament Spacing in Passive Muscle: Effect of Titin-Based Passive Tension","volume":"100","author":"Irving","year":"2011","journal-title":"Biophys. J."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"947","DOI":"10.1002\/jcsm.12319","article-title":"Titin-Based Mechanosensing Modulates Muscle Hypertrophy","volume":"9","author":"Strom","year":"2018","journal-title":"J. Cachexia Sarcopenia Muscle"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e40532","DOI":"10.7554\/eLife.40532","article-title":"Downsizing the Molecular Spring of the Giant Protein Titin Reveals That Skeletal Muscle Titin Determines Passive Stiffness and Drives Longitudinal Hypertrophy","volume":"7","author":"Brynnel","year":"2018","journal-title":"Elife"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1085\/jgp.200509364","article-title":"Isoform Diversity of Giant Proteins in Relation to Passive and Active Contractile Properties of Rabbit Skeletal Muscles","volume":"126","author":"Prado","year":"2005","journal-title":"J. Gen. Physiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1924","DOI":"10.1161\/CIRCULATIONAHA.113.005610","article-title":"Experimentally Increasing Titin Compliance in a Novel Mouse Model Attenuates the Frank-Starling Mechanism but Has a Beneficial Effect on Diastole","volume":"129","author":"Methawasin","year":"2014","journal-title":"Circulation"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"804","DOI":"10.1016\/j.bbamcr.2012.08.011","article-title":"Impact of Titin Isoform on Length Dependent Activation and Cross-Bridge Cycling Kinetics in Rat Skeletal Muscle","volume":"1833","author":"Mateja","year":"2013","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"766","DOI":"10.1038\/nm.2693","article-title":"RBM20, a Gene for Hereditary Cardiomyopathy, Regulates Titin Splicing","volume":"18","author":"Guo","year":"2012","journal-title":"Nat. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"14589","DOI":"10.1073\/pnas.1411493111","article-title":"Deleting Titin\u2019s I-Band\/A-Band Junction Reveals Critical Roles for Titin in Biomechanical Sensing and Cardiac Function","volume":"111","author":"Granzier","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_37","first-page":"435236","article-title":"The Extent of Ventilator-Induced Lung Injury in Mice Partly Depends on Duration of Mechanical Ventilation","volume":"2013","author":"Hegeman","year":"2013","journal-title":"Crit. Care Res. Pract."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1165\/rcmb.2018-0100OC","article-title":"Relation between Respiratory Mechanics, Inflammation, and Survival in Experimental Mechanical Ventilation","volume":"60","author":"Szabari","year":"2019","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2903","DOI":"10.1093\/cvr\/cvab328","article-title":"Titin (TTN): From Molecule to Modifications, Mechanics, and Medical Significance","volume":"118","author":"Loescher","year":"2021","journal-title":"Cardiovasc. Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1038\/nrneurol.2017.148","article-title":"Antisense Oligonucleotides: The next Frontier for Treatment of Neurological Disorders","volume":"14","author":"Rinaldi","year":"2018","journal-title":"Nat. Rev. Neurol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"8952","DOI":"10.1126\/scitranslmed.abe8952","article-title":"Therapeutic Inhibition of RBM20 Improves Diastolic Function in a Murine Heart Failure Model and Human Engineered Heart Tissue","volume":"13","author":"Radke","year":"2021","journal-title":"Sci. Transl. Med."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.jmb.2009.08.009","article-title":"Altered Contractility of Skeletal Muscle in Mice Deficient in Titin\u2019s M-Band Region","volume":"393","author":"Ottenheijm","year":"2009","journal-title":"J. Mol. Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/24\/15689\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:37:55Z","timestamp":1760146675000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/24\/15689"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,10]]},"references-count":42,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["ijms232415689"],"URL":"https:\/\/doi.org\/10.3390\/ijms232415689","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,10]]}}}