High-Throughput Screening for the Discovery of Iron Homeostasis Modulators Using an Extremely Sensitive Fluorescent Probe

• Published in: ACS sensors 2020-09, Vol.5 (9), p.2950-2958
• Main Authors: Hirayama, Tasuku, Niwa, Masato, Hirosawa, Shusaku, Nagasawa, Hideko
• Format: Article
• Language: English
• Subjects: Fluorescent Dyes; High-Throughput Screening Assays; Homeostasis; Iron
• ISSN: 2379-3694; 2379-3694
• DOI: 10.1021/acssensors.0c01445

High-throughput methods for monitoring subcellular labile Fe­(II) are important for conducting studies on iron homeostasis and for the discovery of potential drug candidates for the treatment of iron deficiency or overload. Herein, a highly sensitive and robust fluorescent probe for the detection of intracellular labile Fe­(II) is described. The probe was designed through the rational optimization of the reactivity and responsiveness for an Fe­(II)-induced fluorogenic reaction based on deoxygenation of an N-oxide, which was developed in-house. The probe is ready to use for a 96-well-plate-based high-content imaging of labile Fe­(II) in living cells. Using this simple method, we were able to conduct high-throughput screening of a chemical library containing 3399 compounds. The compound lomofungin was identified as a potential drug candidate for the intracellular enhancement of labile Fe­(II) via a novel mechanism in which the ferritin protein was downregulated.