THE NICD1-CRIP2 AXIS DRIVES GLYCOLYTIC ADAPTATION AND NEURONAL CELL DEATH IN ISCHEMIC STROKE

Ischemic stroke remains a leading cause of morbidity and mortality, characterized by insufficient blood flow that deprives brain tissue of oxygen and glucose. Notch1 is a well-characterized membrane receptor crucial in ischemic conditions. This study investigates the role of the Notch1 intracellular domain (NICD1) and its downstream effects under ischemic conditions.We analyzed the nuclear translocation of NICD1 under oxygen-glucose deprivation (OGD), with redistribution to the cytosol observed during reperfusion. Transcriptomic analysis revealed differentially expressed genes under OGD conditions. Among these, CRIP2 was identified as a key downstream target sensitively regulated by NICD1.NICD1 knockdown resulted in decreased CRIP2 expression at the transcript and protein levels. While p53 also regulates CRIP2, our results suggest that NICD1 plays a more dominant regulatory role under OGD conditions. Furthermore, NICD1 knockdown led to reduced glycolytic enzyme levels including HK2 and PDK1 and lower levels of cleaved caspase-3, underscoring its role in glycolysis and cell death.
These findings were further validated in a tMCAO mouse model, where Crip2 and Hk2 expression were significantly up-regulated in the infarct region. This study provides new insights into the molecular interactions driving ischemic stroke and positions NICD1 and CRIP2 as potential therapeutic targets. Understanding the role of NICD1 and CRIP2 in ischemic conditions could facilitate the development of novel therapeutic strategies aimed at mitigating ischemic stroke–induced brain injury.