Utilizing patient-derived premalignant colon organoid biobank to interrogate existing and novel chemoprevention strategies - Chemoprevention is an important, but challenging field of study due to the lack of pre-cancer model systems for research studies. Familial adenomatous polyposis (FAP), a hereditary condition characterized by polyp formation and a high risk of cancer development, has been the focus of tens of chemoprevention clinical trials all investigating the same target, cyclooxygenase-2 (COX-2). However, due to inconsistent efficacy across studies and significant off-target effects, no agent is currently recommended for FAP chemoprevention. Furthermore, there is limited research on why these agents have failed, leaving a critical gap in understanding that needs to be addressed in order to guide the development of more effective therapeutic strategies. This proposal aims to better understand how signaling and metabolic pathways are altered in premalignant polyps in effort to design more targeted and effective treatments for FAP patients. To facilitate this goal, I have generated a biobank of normal and premalignant organoid samples derived from extensive multi-sampling of individual patients, capturing both inter-polyp and inter-patient heterogeneity. From this, I discovered that polyp organoids derived from the same patient display variable growth rates and differing responses to two COX-2 inhibitors, which could not be rescued by the downstream primary effector PGE2. Moreover, I found polyp organoids exhibit altered metabolism beyond COX-2, which I hypothesize is indicative of a broader metabolic reprogramming that can be explored for future chemoprevention strategies. With the advisory of a team of expert scientists in single cell analyses, proteomics, metabolomics, and CRISPR screens, techniques I will add to my skillset following completion of this proposal, I plan to uncover the mechanisms underlying polyp variable response to COX-2 inhibitors and explore novel metabolic strategies for chemoprevention. In Aim 1 I will investigate the mechanisms of resistance in polyp organoids that show poor responses to COX-2 inhibitors, with the goal of identifying combination therapies to lower toxicity. In Aim 2, I will investigate the mechanisms of COX-2 sensitivity by examining how COX-2 signaling and downstream eicosanoid molecules regulate the activation of signaling cascades that influence cell growth and cell state. Finally, Aim 3 will move beyond COX-2 signaling to identify new metabolic pathways driving cell growth in polyps, providing insight into pre-cancer metabolic rewiring and offering potential new therapeutic strategies. Ultimately, this research has the potential to uncover novel therapeutic strategies that can significantly improve the effectiveness of chemoprevention in FAP patients and pave the way for broader applications in the prevention of sporadic colorectal cancer.
