Author ORCID Identifier
Date Available
12-12-2026
Year of Publication
2024
Document Type
Doctoral Dissertation
Degree Name
Doctor of Philosophy (PhD)
College
Arts and Sciences
Department/School/Program
Chemistry
Advisor
Dr. Christopher I. Richards
Abstract
The mortality rate of cancer establishes it as a leading global health concern, prompting significant investment into cancer research. While the effects of cancer are well known, the specific mechanisms behind therapeutic resistance remain less well-defined. The goal of this study was to develop innovative methods to address current shortcomings in cancer treatment and understanding. To do this, we studied exosome-mimetic nanovesicles as an immunotherapeutic vaccine platform and fluorescence lifetime imaging as a means to measure cancer-associated enzyme activity at the single cell level.
Through the use of a novel method of production, we generated dendritic cell plasma membrane-derived nanovesicles in high yields and leveraged the antigen-presenting and costimulatory properties of dendritic cells for induction of a T cell immune response. We demonstrate that these nanovesicles are able to present antigens in functional immune stimulatory complexes, retaining the parental cell ability to activate CD8+ T cells. Additionally, these nanovesicles were shown to mediate activation of T cells through indirect means. Here, nanovesicles are taken up by bystander dendritic cells, thereby delivering antigen to the dendritic cell and conferring T cell stimulatory capability. Next, we investigated the application of fluorescence lifetime imaging to measure cancer-associated cytochrome P450 enzyme activity at the single-cell level. We demonstrated this approach provides detailed insights into cellular heterogeneity and localized enzyme activity. Additionally, we showed that sensitivity and dynamic range can be tuned to enzyme activity and levels by altering excitation and emission wavelengths.
These advancements offer new and promising avenues to enhance nanoparticle-based immunotherapy and understanding the role of enzyme activity and cellular heterogeneity in cancer progression. Ultimately, the methods developed contribute to improving therapeutic strategies and personalized medicine.
Digital Object Identifier (DOI)
https://doi.org/10.13023/etd.2024.427
Funding Information
This study was supported by the
- Kentucky Pediatric Cancer Research Trust Fund PON2 728 2200002655 (2020-2022)
- National Institutes of Health GM138837 (2021-2022)
- National Institutes of Health GM138882 (2022-2024)
- Craig H. Neilsen Foundation 1000401213 (2022-2024)
Recommended Citation
Harvey, Brock, "Methodological Innovations for Improved Cancer Treatment and Enzyme Activity Insight" (2024). Theses and Dissertations--Chemistry. 199.
https://uknowledge.uky.edu/chemistry_etds/199
