Tracking Mutual Interactions of Mitochondria/Lysosomes/Lipid Droplets in DILI and NAFLD with a Viscosity and Peroxynitrite-Sensitive Single Fluorescent Probe

Drug-induced liver injury and nonalcoholic fatty liver disease are associated with the cellular microenvironment and reactive oxygen species (ROS) levels, which are expected to influence complex and frequent interactions among multiple intracellular organelles. Two stimuli, an AND-gated lipid droplets (LDs), lysosomes, and mitochondria-targeted fluorescent probe (TPAP-Mito) was developed. The probe emits strong fluorescence by stimulation of both mitochondrial peroxynitrite (ONOO^-) and polarity of LDs, or both the ONOO^- and pH of lysosomes. Using a meticulously designed molecular logic gate, it is possible to achieve real-time tracking of three organelles in a synchronized manner, revealing a significant increase in their fusion frequency, mutual contacts, and interactions. This comprehensive study underscores the potential of developing innovative multifunctional, multitargeted probes, which could significantly improve early diagnosis and treatment options for related diseases.