P4‐513: ACTIVATION OF FERROPTOSIS, AN IRON‐DEPENDENT FORM OF NON‐APOPTOTIC DEATH IN NEURONS

Alzheimer's disease (AD) is characterized abnormal accumulation of amyloid-β, tau, neuroinflammation, high oxidative stress, mitochondrial dysfunction, iron deposition, progressive synaptic loss and by severe neuronal loss. However, the mechanisms by which neurons die have remained elusive. The massive neuronal death in AD patients has been linked to apoptosis and necroptosis processes, but recently a novel form of iron-dependent regulated cell death process termed ferroptosis was described and confirmed as an important cell death mechanism for dopaminergic neurons. Some biochemical and morphological features of ferroptosis have been observed in AD: overall alteration of redox homeostasis, marked lipid peroxidation, high amounts of ROS, mitochondrial dysfunction and iron overload. We hypothesize that ferroptosis is activated in AD as a complex mechanism of neuronal death linked with oxidative stress, abnormal iron mobilization, and mitochondrial dysfunction. Differentiated neurons and neuroblastoma cultures were challenged (0 to 200 uM, up to 96 h) with ferroptosis inductors or ferroptosis inductors+inhibitors to determine ferroptotic responses. We employed analytical and advanced imaging techniques, including bioassays, transcriptomics, optical/confocal/electron microscopy and mass spectrometry to determine and compare cell responses, pathways and other features characteristic of ferroptosis. The objective is to determine whether ferroptosis is activated in AD as underlying mechanisms and elucidate the pathways of neuronal loss, identifying new potential molecular targets for interventions related to altered brain homeostasis, oxidative stress, and neuronal death. The approach established, first determine the neuronal responses and mechanisms of ferroptosis in neurodegeneration, using bioassays and high-throughput sequencing of transcriptomics. Also, characterization of morphological features of ferroptotic neurons with advanced analytical imaging techniques, and finally the quantification of ferroptotic biomarkers with mass spectrometry. This project offers a detailed understanding of the molecular mechanisms of iron-dependent cell death in human neurons, identifying key regulatory genes and proteins, unregulated pathways and morphological features of ferroptotic neurons in the brain correlated with neurodegeneration. In addition to generating important basic neuroscience knowledge, it will be possible to identify potential interventions into the altered aging process and translational research that will offer innovative diagnostic and prognostic targets to regulate cell survival and death in neurodegeneration.