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dc.contributor.advisorMcLellan, Alexander
dc.contributor.advisorMuhsin, Morad-Remy
dc.contributor.authorDinh, Thu Nho
dc.date.available2018-11-19T19:54:36Z
dc.date.copyright2018
dc.identifier.citationDinh, T. N. (2018). Disarming tissue factor: generating a model tumour antigen (Thesis, Bachelor of Biomedical Sciences with Honours). University of Otago. Retrieved from http://hdl.handle.net/10523/8606en
dc.identifier.urihttp://hdl.handle.net/10523/8606
dc.description.abstractCoagulative abnormalities are a well-recognised feature of malignancy. The risk of developing thrombotic disorders is increased four-fold in cancer patients and more than six-fold higher in those receiving chemotherapy. Consequently, coagulative disorders are the second leading cause of death in cancer patients. One of the principal mechanisms through which cancer cells promote coagulation is through the upregulation of tissue factor (TF), a transmembrane receptor expressed on cells surrounding the blood vessels and a potent initiator of coagulation. Upon binding to circulating coagulation factor VIIa (FVIIa), TF and FVIIa form a catalytic complex, initiating the extrinsic coagulation pathway and resulting in the rapid formation of blood clots. Chimeric Antigen Receptor (CAR) T cell therapy is an emerging form of immunotherapy which combines the specificity of antibodies with the cytolytic activity of T cells, enabling potent killing of tumours in an antigen-antibody defined interaction. TF has been explored as a therapeutic target for a range of cancer treatments, however its potential as a target antigen for CAR T cell therapy is yet to be realised. The anti-TF monoclonal antibody, TF8-5G9, has been shown to effectively bind to and inhibit the procoagulant function of human TF making it an ideal candidate for a novel CAR structure. The efficacy of the construct would first need to be demonstrated in a pre-clinical setting. However, human TF is highly efficient at complexing with murine FVIIa. Thus, expression of the antigen in a live model could result in major coagulopathies. This research tackles this issue by generating a truncated form of human TF, using inverse PCR, thereby deleting the FVIIa binding sites while disabling any TF-driven coagulation. Flow cytometric and western blot analyses showed that the expression of the truncated TF was mainly intracellular. However, no coagulation was triggered by the low-level surface expression of the truncated TF, as shown by our in vitro coagulation assay. Failure to efficiently export TF to the surface may be attributed to protein misfolding and thus demands different strategies to truncate the TF gene. Together, these results provide the model framework for TF as a novel target in immunotherapy.
dc.format.mimetypeapplication/pdf
dc.language.isoen
dc.publisherUniversity of Otago
dc.rightsAll items in OUR Archive are provided for private study and research purposes and are protected by copyright with all rights reserved unless otherwise indicated.
dc.subjectTissue Factor
dc.subjectCAR T Cells
dc.subjectCancer
dc.subjectImmunotherapy
dc.titleDisarming tissue factor: generating a model tumour antigen
dc.typeThesis
dc.date.updated2018-11-19T03:53:25Z
dc.language.rfc3066en
thesis.degree.disciplineMicrobiology and Immunology
thesis.degree.nameBachelor of Biomedical Sciences with Honours
thesis.degree.grantorUniversity of Otago
thesis.degree.levelHonours
otago.openaccessOpen
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