A clinical need exists for improved methods to identify cancer patients with tumors prone to drug resistance, and prescribe effective alternative therapies. Cellular metabolism is a powerful marker of tumor response to treatment, because the oncogenic drivers targeted by anti-cancer drugs often regulate cellular metabolism. In this dissertation, optical metabolic imaging (OMI) is developed to detect drug-induced metabolism changes in breast cancer cells, in vivo xenograft tumors, and primary-tumor derived organoids. OMI probes the auto-fluorescence intensity and lifetime of NAD(P)H and FAD, coenzymes of metabolism, to quantify protein concentrations and protein-binding dynamics. A composite endpoint, the OMI index, provides a robust, dynamic readout of cellular metabolism. OMI endpoints were evaluated and characterized in a panel of breast cancer cell lines and three xenograft models in vivo with and without drug treatment. For optimal clinical utility, protocols were developed to generate 3D organoid cultures of primary tumor biopsies for high-throughput testing of potential drugs. In xenograft tumor derived organoids, the OMI index decreased in responsive organoids treated with anti-cancer drugs, corroborating with in vivo tumor growth curves. Feasibility experiments on organoids derived from human patient biopsies demonstrated viability of organoids and drug-induced changes in OMI index. The high-resolution images of OMI allow cellular-level analysis of heterogeneity. Heterogeneity profiles of resistant cell populations within organoids were identified and tracked over 72 hours. With these findings, OMI shows potential for development into a high-throughput organoid screen to test the efficacy of a panel of drugs and drug combinations to direct clinical therapy selection and expedite pre-clinical studies.
| Identifer | oai:union.ndltd.org:VANDERBILT/oai:VANDERBILTETD:etd-03092015-132912 |
| Date | 11 March 2015 |
| Creators | Walsh, Alexandra Jule |
| Contributors | H. Charles Manning, Anita Mahadevan-Jansen, Rebecca S. Cook, Melissa C. Skala, Jamey Young |
| Publisher | VANDERBILT |
| Source Sets | Vanderbilt University Theses |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.vanderbilt.edu/available/etd-03092015-132912/ |
| Rights | restrictone, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to Vanderbilt University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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