TGFbeta2 Induces the Formation of Cross-Linked Actin Networks (CLANs) in Human Trabecular Meshwork Cells Through the Smad and Non-Smad Dependent Pathways

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dc.creatorMontecchi-Palmer, Michela
dc.creatorBermudez, Jaclyn Y.
dc.creatorWebber, Hannah C.
dc.creatorPatel, Gaurang C.
dc.creatorClark, Abbot F.
dc.creatorMao, Weiming
dc.creator.orcid0000-0003-3594-6560 (Clark, Abbot F.)
dc.date.accessioned2022-09-13T16:34:18Z
dc.date.available2022-09-13T16:34:18Z
dc.date.issued2017-02
dc.description.abstractPurpose: Increased intraocular pressure results from increased aqueous humor (AH) outflow resistance at the trabecular meshwork (TM) due to pathologic changes including the formation of cross-linked actin networks (CLANs). Transforming growth factor beta2 (TGFbeta2) is elevated in the AH and TM of primary open angle glaucoma (POAG) patients and induces POAG-associated TM changes, including CLANs. We determined the role of individual TGFbeta2 signaling pathways in CLAN formation. Methods: Cultured nonglaucomatous human TM (NTM) cells were treated with control or TGFbeta2, with or without the inhibitors of TGFbeta receptor, Smad3, c-Jun N-terminal kinases (JNK), extracellular signal regulated kinase (ERK), P38, or Rho-associated protein kinase (ROCK). NTM cells were cotreated with TGFbeta2 plus inhibitors for 10 days or pretreated with TGFbeta2 for 10 days followed by 1-hour inhibitor treatment. NTM cells were immunostained with phalloidin-Alexa-488 and 4',6-diamidino-2-phenylindole (DAPI). Data were analyzed using 1-way ANOVA and Dunnett's post hoc test. Results: TGFbeta2 significantly induced CLAN formation (n = 6 to 12, P < 0.05), which was completely inhibited by TGFbeta receptor, Smad3, and ERK inhibitors, as well as completely or partially inhibited by JNK, P38, and ROCK inhibitors, depending on cell strains. One-hour exposure to ROCK inhibitor completely resolved formed CLANs (P < 0.05), whereas TGFbeta receptor, Smad3 inhibitor, and ERK inhibitors resulted in partial or complete resolution. The JNK and P38 inhibitors showed partial or no resolution. Among these inhibitors, the ROCK inhibitor was the most disruptive to the actin stress fibers, whereas ERK inhibition showed the least disruption. Conclusions: TGFbeta2-induced CLANs in NTM cells were prevented and resolved using various pathway inhibitors. Apart from CLAN inhibition, some of these inhibitors also had different effects on actin stress fibers.
dc.description.sponsorshipSupported by National Eye Institute Grant R21EY023048 (W.M.).
dc.identifier.citationMontecchi-Palmer, M., Bermudez, J. Y., Webber, H. C., Patel, G. C., Clark, A. F., & Mao, W. (2017). TGFβ2 Induces the Formation of Cross-Linked Actin Networks (CLANs) in Human Trabecular Meshwork Cells Through the Smad and Non-Smad Dependent Pathways. Investigative ophthalmology & visual science, 58(2), 1288-1295. https://doi.org/10.1167/iovs.16-19672
dc.identifier.issn1552-5783
dc.identifier.issue2
dc.identifier.urihttps://hdl.handle.net/20.500.12503/31732
dc.identifier.volume58
dc.publisherARVO Journals
dc.relation.urihttps://doi.org/10.1167/iovs.16-19672
dc.rights.holderCopyright 2017 The Authors.
dc.rights.licenseAttribution 4.0 International (CC BY 4.0)
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.sourceInvestigative Ophthalmology & Visual Science
dc.subject.meshActins / metabolism
dc.subject.meshAnalysis of Variance
dc.subject.meshAqueous Humor / metabolism
dc.subject.meshBlotting, Western
dc.subject.meshCells, Cultured
dc.subject.meshHumans
dc.subject.meshJNK Mitogen-Activated Protein Kinases / metabolism
dc.subject.meshProtein Serine-Threonine Kinases
dc.subject.meshReceptor, Transforming Growth Factor-beta Type II
dc.subject.meshReceptors, Transforming Growth Factor beta
dc.subject.meshSignal Transduction / physiology
dc.subject.meshSmad3 Protein / metabolism
dc.subject.meshTrabecular Meshwork / drug effects
dc.subject.meshTrabecular Meshwork / metabolism
dc.subject.meshTransforming Growth Factor beta2 / antagonists & inhibitors
dc.subject.meshTransforming Growth Factor beta2 / pharmacology
dc.subject.meshTransforming Growth Factor beta2 / physiology
dc.titleTGFbeta2 Induces the Formation of Cross-Linked Actin Networks (CLANs) in Human Trabecular Meshwork Cells Through the Smad and Non-Smad Dependent Pathways
dc.typeArticle
dc.type.materialtext

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