Design and Characterization of Antibody-Conjugated T-Cells and Mimetic Nanovesicles for Cancer Therapy
Date
2023-12-04
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
ORCID
0000-0002-1869-9986
Type
Thesis
Degree Level
Masters
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.
Description
Keywords
ICTS, CAR-T cells, Ac4ManNAz, DBCO, Click Chemistry, EGFR, ACTs, Antibody conjugated T-cells, Nanovesicles
Citation
Degree
Master of Science (M.Sc.)
Department
Biochemistry
Program
Biochemistry