Mass Spectrometry Reveals that Oxysterols are Secreted from Non-Alcoholic Fatty Liver Disease Induced Organoids

Abstract

Oxysterols are potential biomarkers for liver metabolism that are altered under disease 13 conditions such as non-alcoholic fatty liver disease (NAFLD). We here apply sterolomics to 14 organoids used for disease modeling of NAFLD. Using liquid chromatography-mass spectrometry 15 with on-line sample clean-up and enrichment, we establish that liver organoids produce and 16 secrete oxysterols. We find elevated levels of 26-hydroxycholesterol, an LXR agonist and the 17 first oxysterol in the acidic bile acid synthesis, in medium from steatotic liver organoids 18 compared to untreated organoids. Other upregulated sterols in medium from steatotic liver 19 organoids are dihydroxycholesterols, such as 7α,26–dihydroxycholesterol, and 7α,25- 20 dihydroxycholesterol.