Understanding the impact of the oncogenic signaling network on cellular metabolic network in human breast cancers

We anticipate that oncogenic activation of the PI3K and Ras pathways in breast cancer cells setting will reveal both shared and unique alterations in nucleotide metabolism. While our previous study was focused on the contributions of mTORC1 on nucleotide synthesis, our approach under this proposal, could reveal parallel inputs downstream of these oncogenes that also control specific nucleotide metabolic pathways independently of mTORC1 signaling. This study focuses on alteration of cellular metabolism in a context of oncogenic signaling pathways to uncover liabilities essential for cancer cell growth and proliferation. Through a CRISPR Cas9 screen targeting a panel of key metabolic enzymes representative of all the critical metabolic routes required for cell growth, we hope to uncover metabolic vulnerabilities to significantly sensitize all type of breast cancers to chemotherapeutic agents. Taking advantage of tools for the manipulation of gene expression and advances in read count and length in massively parallel next-generation DNA sequencing (NGS), our objective is to systematically and quantitatively assess the role of the different metabolic network by using metabolic enzyme library of single guide RNA (sgRNA) in the context of oncogenes transformed (PI3K or K-RAS) epithelial breast cell line or non-transformed breast cell line (i.e., MCF10A). The data generated from this proposal will have immediate and important diagnostic and therapeutic implications. Moreover, beyond the translational implications, our data will reveal important insights in the function of cellular metabolic networks in cancer cell biology.