{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,26]],"date-time":"2026-07-26T23:24:51Z","timestamp":1785108291586,"version":"3.55.0"},"reference-count":52,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,7,24]],"date-time":"2024-07-24T00:00:00Z","timestamp":1721779200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Pharmacol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Mirabegron is available for treatment of overactive bladder (OAB). However, mechanisms underlying symptom improvements and long-term effects on bladder smooth muscle cells are uncertain. Contractility and growth of bladder smooth muscle contribute to OAB, and depend on smooth muscle phenotypes, and on muscarinic receptor expression. Here, we examined prolonged exposure to mirabegron (20\u201348\u00a0h) on phenotype markers, muscarinic receptor expression, and phenotype-dependent functions in human bladder smooth muscle cells (hBSMC).<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Expression of markers for contractile (calponin, MYH11) and proliferative (MYH10, vimentin) phenotypes, proliferation (Ki-67), and of muscarinic receptors were assessed by RT-PCR. Proliferation, viability, actin organization and contractions in cultured hBSMC were examined by EdU, CCK-8, phalloidin staining and matrix contraction assays.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Calponin-1 mRNA decreased with 100\u00a0nM and 150\u00a0nM mirabegron applied for 20\u00a0h (0.56\u20130.6 fold of controls). Decreases were resistant to the \u03b2<jats:sub>3<\/jats:sub>-AR antagonist L-748,337 (0.34\u20130.55 fold, 100\u2013150\u00a0nM, 20\u00a0h). After 40\u00a0h, decreases occured in the presence of L-748,337, but not without L-748,337. MYH11 mRNA increased with 150\u00a0nM mirabegron (40\u00a0h, 1.9 fold). This was partly preserved with L-748,337, but not observed after 20\u00a0h mirabegron exposure. Vimentin mRNA reduced with 150\u00a0nM mirabegron after 20\u00a0h, but not after 40\u00a0h, with and without L-748,337 (0.71\u20130.63 fold). MYH10 mRNA expression remained unaffected by mirabegron. Exposure to 150\u00a0nM mirabegron increased Ki-67 mRNA after 20\u00a0h in the presence of, but not without L-748,337, and after 40\u00a0h without, but not with L-748,337. Proliferation rates and actin organization were stable with 50\u2013150\u00a0nM mirabegron (24\u00a0h, 48\u00a0h). Viability increased significantly after mirabegron exposure for 20\u00a0h, and by trend after 40\u00a0h, which was fully sensitive to L-748,337. M2 mRNA was reduced by 20\u00a0h mirabegron, which was resistant to L-748,337. Carbachol (3\u00a0\u00b5M) enhanced time-dependent contractions of hBSMC, which was inhibited by mirabegron (150\u00a0nM) in late phases (24\u00a0h), but not in early phases of contractions.<\/jats:p><jats:p><jats:bold>Conclusion:<\/jats:bold> Mirabegron induces dynamic phenotype alterations and M2 downregulation in hBSMC, which is paralleled by time-shifted anticontractile effects. Phenotype transitions may be involved in improvements of storage symptoms in OAB by mirabegron.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fphar.2024.1446831","type":"journal-article","created":{"date-parts":[[2024,7,24]],"date-time":"2024-07-24T04:22:18Z","timestamp":1721794938000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Dynamic phenotypic shifts and M2 receptor downregulation in bladder smooth muscle cells induced by mirabegron"],"prefix":"10.3389","volume":"15","author":[{"given":"A. E.","family":"Muderrisoglu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Ciotkowska","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B.","family":"Rutz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S.","family":"Hu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S.","family":"Qian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Tamalunas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C. G.","family":"Stief","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Hennenberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,7,24]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"415","DOI":"10.1111\/bph.13367","article-title":"Mirabegron relaxes urethral smooth muscle by a dual mechanism involving \u03b23 -adrenoceptor activation and \u03b11 -adrenoceptor blockade","volume":"173","author":"Alexandre","year":"2016","journal-title":"Br. J. Pharmacol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"363","DOI":"10.1111\/j.1365-201x.1998.tb10694.x","article-title":"Cross-bridge cycling in smooth muscle: a short review","volume":"164","author":"Arner","year":"1998","journal-title":"Acta Physiol. Scand."},{"key":"B3","doi-asserted-by":"publisher","first-page":"1033","DOI":"10.1113\/jphysiol.2006.118828","article-title":"Effects of h1-calponin ablation on the contractile properties of bladder versus vascular smooth muscle in mice lacking SM-B myosin","volume":"577","author":"Babu","year":"2006","journal-title":"J. Physiol."},{"key":"B4","doi-asserted-by":"publisher","first-page":"1114355","DOI":"10.3389\/fcvm.2023.1114355","article-title":"Age-dependent phenotypic modulation of smooth muscle cells in the normal ascending aorta","volume":"10","author":"Balint","year":"2023","journal-title":"Front. Cardiovasc Med."},{"key":"B5","first-page":"649","article-title":"Potent and selective human beta(3)-adrenergic receptor antagonists","volume":"290","author":"Candelore","year":"1999","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"B6","doi-asserted-by":"publisher","first-page":"180","DOI":"10.1186\/s12964-022-00993-2","article-title":"How vascular smooth muscle cell phenotype switching contributes to vascular disease","volume":"20","author":"Cao","year":"2022","journal-title":"Cell. Commun. Signal"},{"key":"B7","unstructured":"Pharmacology\/toxicology NDA\/BLA review and evaluation (NDA 202-611)2012"},{"key":"B8","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1016\/j.yjmcc.2019.01.026","article-title":"Double deletion of calponin 1 and calponin 2 in mice decreases systemic blood pressure with blunted length-tension response of aortic smooth muscle","volume":"129","author":"Feng","year":"2019","journal-title":"J. Mol. Cell. Cardiol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"259","DOI":"10.1159\/000209255","article-title":"Muscarinic receptor subtypes in the lower urinary tract","volume":"83","author":"Giglio","year":"2009","journal-title":"Pharmacology"},{"key":"B10","doi-asserted-by":"publisher","first-page":"207","DOI":"10.1016\/j.eururo.2023.04.008","article-title":"Summary paper on the 2023 European association of urology guidelines on the management of non-neurogenic male lower urinary tract symptoms","volume":"84","author":"Gravas","year":"2023","journal-title":"Eur. Urol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"419","DOI":"10.1016\/s0024-3205(98)00581-5","article-title":"Muscarinic receptor subtypes modulating smooth muscle contractility in the urinary bladder","volume":"64","author":"Hegde","year":"1999","journal-title":"Life Sci."},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1007\/164_2023_678","article-title":"Adrenoceptors in the lower urinary tract","author":"Hennenberg","year":"2023","journal-title":"Handb. Exp. Pharmacol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"180","DOI":"10.1016\/j.bbrc.2015.04.028","article-title":"Myosin heavy chain 10 (MYH10) is required for centriole migration during the biogenesis of primary cilia","volume":"461","author":"Hong","year":"2015","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"B14","doi-asserted-by":"publisher","first-page":"666047","DOI":"10.3389\/fphar.2021.666047","article-title":"Concentration-dependent alpha1-adrenoceptor antagonism and inhibition of neurogenic smooth muscle contraction by mirabegron in the human prostate","volume":"12","author":"Huang","year":"2021","journal-title":"Front. Pharmacol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"176","DOI":"10.1124\/jpet.121.001029","article-title":"Inhibition of full smooth muscle contraction in isolated human detrusor tissues by mirabegron is limited to off-target inhibition of neurogenic contractions","volume":"381","author":"Huang","year":"2022","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"B16","doi-asserted-by":"publisher","first-page":"2525","DOI":"10.1111\/bph.14658","article-title":"beta3 -Adrenoceptors in the normal and diseased urinary bladder-What are the open questions?","volume":"176","author":"Igawa","year":"2019","journal-title":"Br. J. Pharmacol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"2126","DOI":"10.1096\/fj.201901301R","article-title":"Increased expression of desmin and vimentin reduces bladder smooth muscle contractility via JNK2","volume":"34","author":"Javed","year":"2020","journal-title":"FASEB J."},{"key":"B18","doi-asserted-by":"publisher","first-page":"389","DOI":"10.1007\/s00210-008-0365-9","article-title":"Suitability of muscarinic acetylcholine receptor antibodies for immunohistochemistry evaluated on tissue sections of receptor gene-deficient mice","volume":"379","author":"Jositsch","year":"2009","journal-title":"Naunyn Schmiedeb. Arch. Pharmacol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"2144","DOI":"10.1016\/j.clinthera.2012.09.010","article-title":"Pharmacokinetic properties of mirabegron, a \u03b23-adrenoceptor agonist: results from two phase I, randomized, multiple-dose studies in healthy young and elderly men and women","volume":"34","author":"Krauwinkel","year":"2012","journal-title":"Clin. Ther."},{"key":"B20","doi-asserted-by":"publisher","first-page":"17872","DOI":"10.1002\/jcb.29056","article-title":"\u03b2-Adrenoceptor signaling regulates proliferation and contraction of human bladder smooth muscle cells under pathological hydrostatic pressure","volume":"120","author":"Lai","year":"2019","journal-title":"J. Cell. Biochem."},{"key":"B21","doi-asserted-by":"publisher","first-page":"469","DOI":"10.1007\/s12041-018-0930-z","article-title":"Loss of smooth muscle myosin heavy chain results in the bladder and stomach developing lesion during foetal development in mice","volume":"97","author":"Li","year":"2018","journal-title":"J. Genet."},{"key":"B22","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1016\/j.gene.2016.02.040","article-title":"Calponin isoforms CNN1, CNN2 and CNN3: regulators for actin cytoskeleton functions in smooth muscle and non-muscle cells","volume":"585","author":"Liu","year":"2016","journal-title":"Gene"},{"key":"B23","doi-asserted-by":"publisher","first-page":"1357","DOI":"10.1056\/NEJMoa012630","article-title":"Myocardial gene expression in dilated cardiomyopathy treated with beta-blocking agents","volume":"346","author":"Lowes","year":"2002","journal-title":"N. Engl. J. Med."},{"key":"B24","doi-asserted-by":"publisher","first-page":"1862","DOI":"10.1016\/j.biocel.2006.03.014","article-title":"(+)Insert smooth muscle myosin heavy chain (SM-B): from single molecule to human","volume":"38","author":"Low","year":"2006","journal-title":"Int. J. Biochem. Cell. Biol."},{"key":"B25","doi-asserted-by":"publisher","first-page":"1159","DOI":"10.1016\/s0024-3205(01)01488-6","article-title":"Functional muscarinic M2 and M3 receptors and beta-adrenoceptor in cultured rat bladder smooth muscle","volume":"70","author":"Ma","year":"2002","journal-title":"Life Sci."},{"key":"B26","doi-asserted-by":"publisher","first-page":"173287","DOI":"10.1016\/j.ejphar.2020.173287","article-title":"Intermittent \u03b2-adrenergic blockade downregulates the gene expression of \u03b2-myosin heavy chain in the mouse heart","volume":"882","author":"Maccari","year":"2020","journal-title":"Eur. J. Pharmacol."},{"key":"B27","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1002\/nau.20121","article-title":"Altered expression of thin filament-associated proteins in hypertrophied urinary bladder smooth muscle","volume":"25","author":"Mannikarottu","year":"2006","journal-title":"Neurourol. Urodyn."},{"key":"B28","doi-asserted-by":"publisher","first-page":"1089","DOI":"10.1038\/sj.bjp.0706147","article-title":"Muscarinic receptor subtypes in human bladder detrusor and mucosa, studied by radioligand binding and quantitative competitive RT-PCR: changes in ageing","volume":"144","author":"Mansfield","year":"2005","journal-title":"Br. J. Pharmacol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"554","DOI":"10.1124\/pharmrev.121.000523","article-title":"Current and emerging pharmacological targets and treatments of urinary incontinence and related disorders","volume":"75","author":"Michel","year":"2023","journal-title":"Pharmacol. Rev."},{"key":"B30","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1124\/mol.119.118927","article-title":"New author guidelines for displaying data and reporting data analysis and statistical methods in experimental biology","volume":"97","author":"Michel","year":"2020","journal-title":"Mol. Pharmacol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"385","DOI":"10.1007\/s00210-009-0395-y","article-title":"How reliable are G-protein-coupled receptor antibodies?","volume":"379","author":"Michel","year":"2009","journal-title":"Naunyn Schmiedeb. Arch. Pharmacol."},{"key":"B32","doi-asserted-by":"publisher","first-page":"2489","DOI":"10.1007\/s12325-022-02114-4","article-title":"What are realistic expectations to become free of overactive bladder symptoms? Experience from non-interventional studies with propiverine","volume":"39","author":"Muderrisoglu","year":"2022","journal-title":"Adv. Ther."},{"key":"B33","doi-asserted-by":"publisher","first-page":"596","DOI":"10.1016\/j.eururo.2017.12.031","article-title":"EAU guidelines on assessment and nonsurgical management of urinary incontinence","volume":"73","author":"Nambiar","year":"2018","journal-title":"Eur. Urol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"366","DOI":"10.1080\/00365590500192918","article-title":"Transforming growth factor-beta1 regulates cell growth and causes downregulation of SMemb\/non-muscle myosin heavy chain B mRNA in human prostate stromal cells","volume":"39","author":"Obara","year":"2005","journal-title":"Scand. J. Urol. Nephrol."},{"key":"B35","doi-asserted-by":"publisher","first-page":"118","DOI":"10.1016\/j.eururo.2013.03.004","article-title":"EAU guidelines on the treatment and follow-up of non-neurogenic male lower urinary tract symptoms including benign prostatic obstruction","volume":"64","author":"Oelke","year":"2013","journal-title":"Eur. Urol."},{"key":"B36","doi-asserted-by":"publisher","first-page":"397","DOI":"10.1007\/s00210-009-0393-0","article-title":"Lack of specificity of commercially available antisera against muscarinergic and adrenergic receptors","volume":"379","author":"Pradidarcheep","year":"2009","journal-title":"Naunyn Schmiedeb. Arch. Pharmacol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"1280","DOI":"10.1002\/jcb.20016","article-title":"Proliferation marker pKi-67 occurs in different isoforms with various cellular effects","volume":"91","author":"Schmidt","year":"2004","journal-title":"J. Cell. Biochem."},{"key":"B38","doi-asserted-by":"publisher","first-page":"1148","DOI":"10.1124\/jpet.103.063735","article-title":"Signal transduction underlying carbachol-induced contraction of human urinary bladder","volume":"309","author":"Schneider","year":"2004","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"B39","doi-asserted-by":"publisher","first-page":"375","DOI":"10.1007\/978-3-642-23274-9_16","article-title":"Muscarinic agonists and antagonists: effects on the urinary bladder","author":"Sellers","year":"2012","journal-title":"Handb. Exp. Pharmacol."},{"key":"B40","doi-asserted-by":"publisher","first-page":"271","DOI":"10.1007\/s004410000262","article-title":"Increased expression of non-muscle myosin heavy chain-B in connective tissue cells of hypertrophic rat urinary bladder","volume":"304","author":"Sjuve","year":"2001","journal-title":"Cell. Tissue Res."},{"key":"B41","doi-asserted-by":"publisher","first-page":"C869","DOI":"10.1152\/ajpcell.00154.2007","article-title":"Intermediate filaments in smooth muscle","volume":"294","author":"Tang","year":"2008","journal-title":"Am. J. Physiol. Cell. Physiol."},{"key":"B42","doi-asserted-by":"publisher","first-page":"381","DOI":"10.1016\/j.ejps.2012.03.001","article-title":"An aryloxypropanolamine h\u03b23-adrenoceptor agonist as bladder smooth muscle relaxant","volume":"46","author":"Tasler","year":"2012","journal-title":"Eur. J. Pharm. Sci."},{"key":"B43","doi-asserted-by":"publisher","first-page":"1182","DOI":"10.1016\/j.theriogenology.2016.04.008","article-title":"The role of adrenergic activation on murine luteal cell viability and progesterone production","volume":"86","author":"Wang","year":"2016","journal-title":"Theriogenology"},{"key":"B44","doi-asserted-by":"publisher","first-page":"L240","DOI":"10.1152\/ajplung.00199.2006","article-title":"Dissociation of Crk-associated substrate from the vimentin network is regulated by p21-activated kinase on ACh activation of airway smooth muscle","volume":"292","author":"Wang","year":"2007","journal-title":"Am. J. Physiol. Lung Cell. Mol. Physiol."},{"key":"B45","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1016\/j.yjmcc.2013.04.025","article-title":"Anti-hypertrophic and anti-oxidant effect of beta3-adrenergic stimulation in myocytes requires differential neuronal NOS phosphorylation","volume":"62","author":"Watts","year":"2013","journal-title":"J. Mol. Cell. Cardiol."},{"key":"B46","doi-asserted-by":"publisher","first-page":"432","DOI":"10.1159\/000430366","article-title":"Contribution of Nrf2 to atherogenic phenotype switching of coronary arterial smooth muscle cells lacking CD38 gene","volume":"37","author":"Xu","year":"2015","journal-title":"Cell. Physiol. Biochem."},{"key":"B47","doi-asserted-by":"publisher","first-page":"201","DOI":"10.1124\/jpet.120.000301","article-title":"Possible involvement of muscarinic receptor blockade in mirabegron therapy for patients with overactive bladder","volume":"377","author":"Yamada","year":"2021","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"B48","doi-asserted-by":"publisher","first-page":"634","DOI":"10.1016\/j.bbrc.2016.11.011","article-title":"Beta-adrenergic signaling affect osteoclastogenesis via osteocytic MLO-Y4 cells' RANKL production","volume":"488","author":"Yao","year":"2017","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"B49","doi-asserted-by":"publisher","first-page":"e021889","DOI":"10.1136\/bmjopen-2018-021889","article-title":"Real-world persistence and adherence to oral antimuscarinics and mirabegron in patients with overactive bladder (OAB): a systematic literature review","volume":"8","author":"Yeowell","year":"2018","journal-title":"BMJ Open"},{"key":"B50","doi-asserted-by":"publisher","first-page":"H1012-H1018","DOI":"10.1152\/ajpheart.00329.2018","article-title":"Proteomic analysis of aortic smooth muscle cell secretions reveals an association of myosin heavy chain 11 with abdominal aortic aneurysm","volume":"315","author":"Yokoyama","year":"2018","journal-title":"Am. J. Physiol. Heart Circ. Physiol."},{"key":"B51","doi-asserted-by":"publisher","first-page":"119365","DOI":"10.1016\/j.lfs.2021.119365","article-title":"Glutamine switches vascular smooth muscle cells to synthetic phenotype through inhibiting miR-143 expression and upregulating THY1 expression","volume":"277","author":"Zhang","year":"2021","journal-title":"Life Sci."},{"key":"B52","doi-asserted-by":"publisher","first-page":"65","DOI":"10.1007\/s11010-017-3136-4","article-title":"CD137-CD137L interaction modulates neointima formation and the phenotype transformation of vascular smooth muscle cells via NFATc1 signaling","volume":"439","author":"Zhong","year":"2018","journal-title":"Mol. Cell. Biochem."}],"container-title":["Frontiers in Pharmacology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2024.1446831\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,7,24]],"date-time":"2024-07-24T04:22:23Z","timestamp":1721794943000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2024.1446831\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,7,24]]},"references-count":52,"alternative-id":["10.3389\/fphar.2024.1446831"],"URL":"https:\/\/doi.org\/10.3389\/fphar.2024.1446831","relation":{},"ISSN":["1663-9812"],"issn-type":[{"value":"1663-9812","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,7,24]]},"article-number":"1446831"}}