Effect of Microbial Short-Chain Fatty Acids on CYP3A4-Mediated Metabolic Activation of Human Pluripotent Stem Cell-Derived Liver Organoids

Abstract

The early and accurate prediction of the hepatotoxicity of new drug targets during nonclinical drug development is important to avoid postmarketing drug withdrawals and late-stage failures. We previously established long-term expandable and functional human-induced pluripotent stem cell (iPSC)-derived liver organoids as an alternative source for primary human hepatocytes. However, PSC-derived organoids are known to present immature fetal characteristics. Here, we treated these liver organoids with microbial short-chain fatty acids (SCFAs) to improve metabolic maturation based on microenvironmental changes in the liver during postnatal development. The effects of the three main SCFA components (acetate, propionate, and butyrate) and their mixture on liver organoids were determined. Propionate (1 µM) significantly promoted the CYP3A4/CYP3A7 expression ratio, and acetate (1 µM), propionate (1 µM), and butyrate (1 µM) combination treatment, compared to no treatment (control), substantially increased CYP3A4 activity and albumin secretion, as well as gene expression. More importantly, mixed SCFA treatment accurately revealed troglitazone-induced hepatotoxicity, which was redeemed on a potent CYP3A4 inhibitor ketoconazole treatment. Overall, we determined, for the first time, that SCFA mixture treatment might contribute to the accurate evaluation of the CYP3A4-dependent drug toxicity by improving metabolic activation, including CYP3A4 expression, of liver organoids.

Investigation of Gynura segetum root extract (GSrE) induced hepatotoxicity based on metabolomic signatures and microbial community profiling in rats

“…For example, butyrate has a detoxification effect on a genipin-induced liver injury by promoting colonic integrity and promoting Nrf2 activation (Luo et al, 2021 ). A study by Mun et al ( 2021 ) showed that an SCFA mixture treatment improved metabolic activation of liver organoids and contributed to the accurate evaluation of the CYP3A4-dependent drug toxicity. In our metabolomics results, the level of pentanoic acid in urine was decreased after GSrE-induced hepatotoxicity, and the level on day 21 was lower than that on day 10.…”

Liver organoids: From fabrication to application in liver diseases

“…The authors embedded foregut globules in a stromal gel, followed by co-culture with retinoic acid (RA) to directionally differentiate PSCs into foregut-derived organoids, thereby generating more complete organoids containing both epithelial components and mesenchymal cells which can differentiate into supporting lineages. Recently, in response to the disadvantage that PSC-derived organoids exhibit only immature fetal characteristics, Mun et al (2021) used microbial short-chain fatty acids (SCFAs) to simulate changes in the microenvironment of the liver during postnatal development. This approach could increase albumin secretion and P450 activity, as well as the expression of hepatocyte genes ( Aloia et al, 2019 ), which improved the metabolic ripening of IPSC-derived liver organoids.…”

Advanced human liver models for the assessment of drug-induced liver injury

“…Hepatocytes in pericentral zone 3 express high levels of CYP enzymes and execute xenobiotic metabolism. Artificial hepatic zonation via a Wnt gradient represents zone 3-specific APAP toxicity that forms toxic reactive metabolites through CYP2E1, CYP1A2, and CYP3A4 [5,[37][38][39][40]. Furthermore, inter-organ interactions can be simulated using this bioengineering technology, which mirrors absorption (gut), distribution, metabolism (liver), and excretion (kidney) [34].…”