Nutrient availability is often compromised within the tumour microenvironment (TME), with concentrations of critical molecules, including glucose and amino acids, falling below the levels typically required to support cell proliferation. Despite these challenges, cancer cells are able to survive and thrive within the TME. While cancer cells are subjected to chronic nutrient stress within the TME, previous studies have largely focused on characterising the cellular response to acute nutrient deprivation. In this project, we have utilised physiologically relevant cell culture media to develop novel models of acute and chronic nutrient deprivation. Interestingly, while acute and chronic nutrient deprivation induce widespread transcriptional changes in cancer cells, unique gene expression changes are observed between the two conditions. Notably, enrichment of signatures associated with the stress-responsive transcription factor Activating Transcription Factor 4 (ATF4) are observed in response to chronic nutrient deprivation, but not in response to acute deprivation. Employing ATF4 knockdown cancer cell lines, we are investigating the impact of ATF4 disruption on the cellular response to nutrient deprivation. Moreover, unbiased CRISPR screening approaches will identify genes required for cell survival during nutrient stress. Future studies will investigate the impact of nutrient deprivation on the ability of cells to overcome other microenvironmental stressors including exposure to chemotherapy/targeted therapy agents. Through understanding the molecular mechanisms that cancer cells employ to adapt to changes in microenvironmental nutrient availability we hope to identify dependencies or vulnerabilities that can be targeted for cancer therapy.