Colorectal cancer (CRC) remains a public health problem worldwide, with dietary factors being the leading cause of CRC-related death. Growing evidence suggests that diet and nutrition promote tumor initiation and progression by altering stem cell metabolism. Therefore, identifying metabolic pathways that determine stem cell activity in CRC may offer a new category of therapeutic targets. Here, we propose to perform a retrospective analysis for single-cell RNA-seq data of 34 CRC patients to find potential candidate genes encoding rate-limiting enzymes of major metabolic pathways that are differentially expressed in the stem cell clusters between normal and tumor tissue. Our preliminary analyses indicate that HMGCS2, and to a lesser extent, CPT1A, are significantly upregulated in stem cells in normal tissue. On the other hand, FASN, and to a lesser degree, G6PD, PFKM, and PPAT, are upregulated in stem cells in tumor tissue. We will further evaluate the tumor-suppressive or pro-tumorigenic potential of the identified metabolic genes by performing a functional assay using organoid culture systems. Specifically, we will introduce genetic perturbations and metabolic rescuing to examine the regulatory role of the identified metabolic pathways. This project aims to identify key metabolic pathways that critically determine the tumorigenic activity of the stem cells in colorectal cancer. We expect the identified metabolic genes to serve as diagnostic and prognostic markers, the genetically modified organoids to be used as disease models for drug screening, and the biomedical agents for metabolic rescuing to inspire nutraceutical strategies for preventing or treating CRC.