A single fluorescent probe to examine the dynamics of mitochondria-lysosome interplay and extracellular vesicle role in ferroptosis.

Ferroptosis is a non-apoptotic form of cell death characterized by iron-dependent lipid peroxidation and glutathione (GSH) depletion. Despite recent advances, challenges remain in understanding the bidirectional interactions or interplay between organelles during ferroptosis. In this study, we aimed to understand the interplay between mitochondria (Mito) and lysosomes (Lyso) in cell homeostasis and ferroptosis. For this purpose, we designed a single fluorescent probe that marks GSH in Mito and hypochlorous acid (HOCl) in Lyso with two distinct emissions. Using this dual-targeted single fluorescent probe (9-morphorino pyronine), we detected Mito-Lyso interplay in ferroptosis. We disclosed differences in Mito-Lyso interplay depending on the induction of ferroptosis. Although erastin treatment decreased GSH, RSL3 triggered a HOCl burst, and FIN56- and FINO2-induced ferroptosis increased GSH and HOCl. Additionally, we showed that only extracellular vesicles generated during erastin-induced ferroptosis could spontaneously move and dock to neighboring cells, resulting in accelerated cell death. [Display omitted] • Simultaneous tracking of the GSH level in mitochondria (Mito) and the HOCl level in lysosomes (Lyso), respectively • Detected distinct Mito-Lyso interplay dynamics and GSH-HOCl crosstalk in ferroptosis • Diverse extracellular vesicle (EV) dynamics in different ferroptosis pathways • Spontaneous movement and docking of the EV accelerate neighboring cell death Wang et al. show the different dynamics of mitochondria-lysosome interplay in the four ferroptosis pathways. They identify diverse extracellular vesicles (EVs) in ferroptosis, of which only erastin-induced EVs can spontaneously move and dock to neighboring cells, resulting in accelerated cell death. [ABSTRACT FROM AUTHOR]