Investigating the Stimulator of Interferon Genes Pathway as a Translational Immunogenic Therapy for Soft Tissue Sarcoma

Hildebrand, Karys Maddison

Investigating the Stimulator of Interferon Genes Pathway as a Translational Immunogenic Therapy for Soft Tissue Sarcoma

dc.contributor.advisor	Monument, Michael James
dc.contributor.author	Hildebrand, Karys Maddison
dc.contributor.committeemember	Jirik, Frank
dc.contributor.committeemember	Muruve, Daniel
dc.contributor.committeemember	Dufour, Antoine
dc.date.accessioned	2024-06-18T15:57:03Z
dc.date.available	2024-06-18T15:57:03Z
dc.date.issued	2024-06-17
dc.description.abstract	Undifferentiated pleomorphic sarcoma (UPS) is one of the most common, aggressive, and metastatic soft tissue sarcomas in adults. Generally, UPS are unresponsive to conventional chemotherapies or immunotherapies, which is believed to be attributed to the immunosuppressive tumor microenvironment (TME) of these malignancies. Preclinical studies in the murine KP model of UPS showed that intratumoral (i.t.) activation of the STimulator of INterferon Genes (STING) pathway using the murine STING agonist DMXAA, elicited immune mediated UPS clearance in 50-75% of treated mice. To assess the translational potential of STING immunotherapy, I tested the anti-tumor efficacy of three STING agonists in the KP model of UPS capable of activating both human and murine STING. Excitingly, E7766 emerged as a translational STING agonist which can shift the UPS TME towards an immunologically inflamed phenotype. Thirty-eight percent of E7766 treated mice eradicated their primary tumors which was CD8+ T-cell dependent. Of the mice that eradicated primary UPS tumors, 87.5% develop protective immunity against UPS re-challenge. Next, I investigated which cell types in the UPS TME engage in STING signaling following therapy. I found that STING expression in non-malignant host cells and not UPS cells is required to observe tumor eradication following STING immunotherapy. Single cell RNA sequencing of UPS tumors revealed that neutrophils are abundant cells in the TMEs of UPS tumors treated with DMXAA and E7766 at both timepoints. Myeloid cells were identified as the cell type with the highest interferon stimulated gene (ISG) score. Both DMXAA and E7766 maintain higher ISG scores in myeloid and lymphoid cells relative to control and CDN at the 1-week timepoint. Finally, I developed a novel gene therapy tool to explore forced expression of the constitutively active mutant hSTINGN154S protein using plasmid DNA. Using these tools, I transfected TAO1 UPS and HEK293T cells and confirmed the functional status of the hSTINGN154S protein’s expression in vitro. In summary, these data suggest that E7766 is an exciting therapeutic candidate for UPS, and further investigation into the importance and consequences of STING signaling in various UPS TME cell types is required to understand therapeutic mechanisms of this therapy.
dc.identifier.citation	Hildebrand, K. M. (2024). Investigating the stimulator of interferon genes pathway as a translational immunogenic therapy for soft tissue sarcoma (Doctoral thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.
dc.identifier.uri	https://hdl.handle.net/1880/118982
dc.language.iso	en
dc.publisher.faculty	Cumming School of Medicine
dc.publisher.institution	University of Calgary
dc.rights	University 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.subject	Stimulator of interferon genes
dc.subject	Undifferentiated pleomorphic sarcoma
dc.subject	Soft tissue sarcoma
dc.subject	Immunotherapy
dc.subject	Tumor microenvironment
dc.subject.classification	Oncology
dc.subject.classification	Immunology
dc.title	Investigating the Stimulator of Interferon Genes Pathway as a Translational Immunogenic Therapy for Soft Tissue Sarcoma
dc.type	doctoral thesis
thesis.degree.discipline	Medicine – Medical Sciences
thesis.degree.grantor	University of Calgary
thesis.degree.name	Doctor of Philosophy (PhD)
ucalgary.thesis.accesssetbystudent	I do not require a thesis withhold – my thesis will have open access and can be viewed and downloaded publicly as soon as possible.