Design and Characterization of Antibody-Conjugated T-Cells and Mimetic Nanovesicles for Cancer Therapy
| dc.contributor.advisor | Geyer, Ron | |
| dc.contributor.committeeMember | Luo, Yu | |
| dc.contributor.committeeMember | Lukong, Erique | |
| dc.contributor.committeeMember | Uppalapati , Maruti | |
| dc.contributor.committeeMember | Price, Eric | |
| dc.creator | Medina Salguero, Andres X | |
| dc.creator.orcid | 0000-0002-1869-9986 | |
| dc.date.accessioned | 2023-12-04T20:54:28Z | |
| dc.date.available | 2023-12-04T20:54:28Z | |
| dc.date.copyright | 2023 | |
| dc.date.created | 2023-11 | |
| dc.date.issued | 2023-12-04 | |
| dc.date.submitted | November 2023 | |
| dc.date.updated | 2023-12-04T20:54:28Z | |
| dc.description.abstract | The development of immune cell therapies (ICTs) such as chimeric antigen receptor T (CAR-T) cells and bi-specific T cell engagers (BiTEs) are revolutionizing cancer treatment. However, production of these therapies is challenging, tedious, and costly. To overcome these obstacles, we are developing a more efficient and cost-effective ICT that does not require genetic engineering of T cells. This therapy involves metabolically engineering T cells to incorporate tetraacetylated N-azidoacetyl-D-mannosamine (Ac4ManNAz) into cell surface proteins, allowing dibenzocyclooctyne (DBCO)-labelled antibodies to be conjugated on the cell surface using a strain-promoted alkyne-azide cycloaddition (SPAAC) reaction. The conjugation process does not depend on the identity of the antibody, enabling antibody-conjugated T cells (ACTs) to be personalized with one or more antibodies, according to their intended application. In this study, we engineered T cells by conjugating them to the epidermal growth factor receptor (EGFR) antibody nimotuzumab. Nimotuzumab-conjugated ACTs interacted better with EGFR-positive cell lines and enhanced the killing efficacy compared to unmodified T cells. Current ICTs such as CAR-T cells have a limited efficacy against solid tumors. To overcome this limitation, we developed a second strategy to target and kill tumor cells. We produced mimetic nanovesicles (M-NVs) from activated-T cells. The small size of M-NVs should allow them to better penetrate solid tumors. M-NVs were labelled with 6-Azidohexanoic acid NHS Ester (NHS-AZ) followed by DBCO-nimotuzumab conjugation. Nimotuzumab-conjugated M-NVs inhibited EGFR-positive cancer cell growth better than non-targeted M-NVs. In summary, our strategy to construct nimotuzumab-conjugated ACTs and M-NVs in a simple, robust, and cost-effective manner allows for an adaptive platform that is translatable to other research labs, being a promising strategy to enhance cancer immune therapies. | |
| dc.format.mimetype | application/pdf | |
| dc.identifier.uri | https://hdl.handle.net/10388/15314 | |
| dc.language.iso | en | |
| dc.subject | ICTS, CAR-T cells, Ac4ManNAz, DBCO, Click Chemistry, EGFR, ACTs, Antibody conjugated T-cells, Nanovesicles | |
| dc.title | Design and Characterization of Antibody-Conjugated T-Cells and Mimetic Nanovesicles for Cancer Therapy | |
| dc.type | Thesis | |
| dc.type.material | text | |
| thesis.degree.department | Biochemistry | |
| thesis.degree.discipline | Biochemistry | |
| thesis.degree.grantor | University of Saskatchewan | |
| thesis.degree.level | Masters | |
| thesis.degree.name | Master of Science (M.Sc.) |