Identifying Compounds that Alleviate Acute Hypoxic Stress by Fast-tracking Cellular Adaptation

Severe acute hypoxic stress is a major contributor to the pathology of human diseases, including ischemic disorders. Current treatments focus on managing consequences of hypoxia, with few addressing cellular adaptation to low-oxygen environments. Here, we hypothesize that accelerating hypoxia adaptation could provide a strategy to alleviate acute hypoxic stress. We developed a high-content phenotypic screening platform to identify compounds that fast-track adaptation to hypoxic stress. Our platform captures a high-dimensional phenotypic hypoxia response trajectory consisting of normoxic, acutely stressed, and chronically adapted cell states. Leveraging this trajectory, we identify compounds that phenotypically shift cells from the acutely stressed towards the adapted state, revealing mTOR/PI3K or BET inhibition as strategies to induce this phenotypic shift. Importantly, our compound hits promote the survival of liver cells exposed to ischemia-like stress, and rescue cardiomyocytes from hypoxic stress. Our platform offers a general, target-agnostic strategy to accelerate cellular adaptation, applicable across various stress conditions. Overall design: RNA-seq profiling of HepG2 cells under 3 oxygen conditions: Normoxia (21% oxygen), 24hr treatment of hypoxia (1% oxygen), and 6day treatment of hypoxia (1% oxygen).