Abstract Despite cancer impressive strides in conventional small molecule therapeutics and novel cancer immunotherapies, continues to be a devastating disease. Certain cancers are refractory to available therapies, and patients' responses show large individual variability. Cancer involves striking dysregulation of epigenetic pathways, with pharmacological approaches targeting chromatin regulators in the clinic and under development. There is also profound involvement of metabolic pathways in cancer, including fascinating nuclear-localized metabolic enzymes. These paradigm-shifting pathways represent an entirely new avenue for targeted therapies, with the potential to directly and specifically modulate cancer-related gene expression programs. However, our understanding of epigenetic mechanisms in cancer, especially as they connect to nuclear metabolic pathways, remains in its infancy. Our history of groundbreaking research revealing new chromatin biology and uncovering genomics and proteomics of the transcription factor p53, both in its role as a crucial tumor suppressor and as a ruinous oncogene, underpins our proposed novel directions. In addition, we have new findings of a chromatin-localized role of a cancer-linked metabolic enzyme directly “fueling” a histone modifying enzyme. Utilizing this background, in this revised proposal, we propose to investigate novel epigenetic regulation and its intersection with nuclear metabolism, using a variety of normal and cancer cell lines, as well as translational mouse models of cancer. We will investigate pivotal developmental- and disease-relevant DNA regulatory elements, called enhancers, which our published findings expose as crucial in p53 function, but which remain understudied in both wildtype and mutant p53 function. In addition, we have recently illuminated a with will speckles chromatin, expression. enzyme wholly novel and unanticipated mechanism of wildtype p53 associating membrane-less bodies, called nuclear speckles, to traffic p53's gene targets for enhanced expression; we dive deeply into the underlying mechanisms for wildtype and oncogenic p53, and explore alterations of in cancer. Other recent findings implicate a nuclear metabolic-epigenetic axis to coordinate, directly at metabolic enzyme production of cofactors with chromatin enzyme function to activate gene We will unravel mechanisms underlying this organization, identify new examples of metabolic coordination with epigenetic enzymes, and determinewhether the nuclear metabolic-epigenetic axis is critical to cancer. Crucially, we have developed an inhibitor targeting ACSS2, a nuclear metabolic enzyme, which presages potential new advancement in therapy. Taken together, our combined focus on roles and interactions of epigenetics and metabolism related to cancer promises to delineate novel mechanisms involved in tumor formation. This paradigm-shifting, multidisciplinary work will bridge separate but related areas of cancer biology and ...