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dc.contributor.advisorDeans, Julie
dc.contributor.authorSharma, Ritu
dc.date.accessioned2017-02-02T18:15:04Z
dc.date.available2017-02-02T18:15:04Z
dc.date.issued2017-01
dc.date.submittedJanuary 2017en
dc.identifier.citationSharma, R. (2017). The Role of Endothelial Focal Adhesion Kinase (FAK) and FAK Related Non-Kinase (FRNK) in Leukocyte Recruitment (Unpublished master's thesis). University of Calgary, Calgary, AB. doi:10.11575/PRISM/28323en_US
dc.identifier.urihttp://hdl.handle.net/11023/3640
dc.description.abstractEndothelial cells form the first barrier that leukocytes have to breach in order to reach the site of inflammation. Focal adhesions are the cellular structures that connect the endothelial cytoskeleton to the extracellular matrix. Focal adhesion kinase (FAK) is a signalling protein that is localized to focal adhesions and becomes phosphorylated during leukocyte recruitment. The main aim of this thesis research was to test the hypothesis that FAK is required for leukocyte transmigration. We used in vitro models of TNF-α - and IL-4-mediated inflammation to examine the recruitment and transmigration of neutrophils and eosinophils, respectively. We observed shear-independent loss of FAK in the proximity of neutrophil transmigration. Downregulation of FAK by siRNA, or disruption of FAK signalling by overexpression of FRNK (FAK-related non kinase, an inhibitor of FAK) decreased neutrophil transmigration without interfering with the TNF-α signalling pathway, implicating a functional role for FAK during neutrophil transmigration. In contrast, downregulation of FAK had no effect on eosinophil transmigration. Surprisingly, FRNK overexpression reduced eosinophil transmigration and this was shown to occur independently of FAK. FRNK blocked eosinophil recruitment and transmigration by preventing transcription and protein expression of IL-4 mediated VCAM-1 and CCL26. Interestingly, the FAK inhibitor FAK-II also blocked VCAM-1 and CCL26 expression through an unknown mechanism that we propose involves enhanced availability of the scaffolding domains of inactivated FAK. Finally, we attempted to determine how FRNK regulates expression of VCAM-1. Phosphorylation and nuclear localization of the transcription factor STAT6 were unaffected by FRNK. We found that, in addition to STAT6, IL-4 also regulates expression of GATA6, which induced VCAM-1 expression, and that loss of GATA6 attenuated eosinophil recruitment and transmigration. FRNK reduced IL-4-induced transcription of GATA6 but not its translation. Although we were unable to determine how FRNK regulates VCAM-1, endogenous FRNK was shown to be regulated by IL-4, suggesting an anti-inflammatory role for FRNK in an IL-4 model of leukocyte recruitment.en_US
dc.language.isoeng
dc.rightsUniversity of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.
dc.subjectEducation--Sciences
dc.titleThe Role of Endothelial Focal Adhesion Kinase (FAK) and FAK Related Non-Kinase (FRNK) in Leukocyte Recruitment
dc.typemaster thesis
dc.publisher.facultyGraduate Studies
dc.publisher.institutionUniversity of Calgaryen
dc.identifier.doihttp://dx.doi.org/10.11575/PRISM/28323
thesis.degree.nameMaster of Science
thesis.degree.nameMS
thesis.degree.nameMSc
thesis.degree.disciplineImmunology
thesis.degree.grantorUniversity of Calgary
atmire.migration.oldid5298
dc.contributor.committeememberDeVinney, Rebekah
dc.contributor.committeememberMuruve, Daniel
dc.publisher.placeCalgaryen


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