Identification of indocyanine green as a STT3B inhibitor against mushroom α-amanitin cytotoxicity

• Published in: Nature communications 2023-05, Vol.14 (1), p.2241-15, Article 2241
• Main Authors: Wang, Bei, Wan, Arabella H., Xu, Yu, Zhang, Ruo-Xin, Zhao, Ben-Chi, Zhao, Xin-Yuan, Shi, Yan-Chuan, Zhang, Xiaolei, Xue, Yongbo, Luo, Yong, Deng, Yinyue, Neely, G. Gregory, Wan, Guohui, Wang, Qiao-Ping
• Format: Article
• Language: English
• Subjects: 14/34; 38/43; 49/47; 631/154/1435/2418; 631/154/570; 692/308/153; 82/1; Alpha-Amanitin - pharmacology; Amanita; Amanitin; Animals; Antidotes; Biocompatibility; Biosynthesis; Blocking; Cell lines; CRISPR; Cytotoxicity; Death; Drug screening; Fatalities; Genomes; Glycan; Hepatocytes; Hexosyltransferases; Humanities and Social Sciences; Humans; In vivo methods and tests; Indocyanine Green - pharmacology; Inhibitors; Liver; Male; Membrane Proteins; Mice; Mortality; multidisciplinary; Mushroom poisoning; Mushrooms; Mycotoxins; Organoids; Poisons; Science; Science (multidisciplinary); Toxicity; Toxins
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-023-37714-3

The “death cap”, Amanita phalloides , is the world’s most poisonous mushroom, responsible for 90% of mushroom-related fatalities. The most fatal component of the death cap is α-amanitin. Despite its lethal effect, the exact mechanisms of how α-amanitin poisons humans remain unclear, leading to no specific antidote available for treatment. Here we show that STT3B is required for α-amanitin toxicity and its inhibitor, indocyanine green (ICG), can be used as a specific antidote. By combining a genome-wide CRISPR screen with an in silico drug screening and in vivo functional validation, we discover that N-glycan biosynthesis pathway and its key component, STT3B, play a crucial role in α-amanitin toxicity and that ICG is a STT3B inhibitor. Furthermore, we demonstrate that ICG is effective in blocking the toxic effect of α-amanitin in cells, liver organoids, and male mice, resulting in an overall increase in animal survival. Together, by combining a genome-wide CRISPR screen for α-amanitin toxicity with an in silico drug screen and functional validation in vivo, our study highlights ICG as a STT3B inhibitor against the mushroom toxin. There is currently no specific antidote for death cap mushroom poisoning treatment. Here, the authors identify STT3B as a druggable target and show that indocyanine green is a STT3B inhibitor that can block α-amanitin toxicity in cell lines, liver organoids and mice.