Predictive approaches to mitigate risk of hepatotoxicity of drugs in children - ABSTRACT Understanding and mitigating drug-induced liver injury (DILI) in children is crucial for pediatric healthcare. Children are more prone to liver damage due to differences in drug metabolism, liver development, and unique susceptibilities. DILI can cause immediate health risks and long-term issues like liver failure or chronic liver disease. Since children often need medications for chronic diseases such as epilepsy, infections, and pain, ensuring the safety of drugs is vital. Addressing DILI in children through advanced methods not only improves drug safety but also supports personalized medicine, enhancing care quality and reducing adverse drug reactions. For example, acetaminophen, commonly used for pain and fever, can cause severe liver damage. Valproic acid, used for epilepsy, also poses a risk of liver toxicity, especially with long-term use. In this proposal, we would like to investigate mechanisms of toxicity of acetaminophen, valproic acid, amoxicillin- clavulanate, cannabidiol (CBD), and isoniazid, which covers critical pharmaceuticals in pediatric healthcare, and develop biomarkers of pediatric DILI caused by these pharmaceuticals. The proposal has two main aims. Aim 1 is to evaluate the effect of selected drugs on liver toxicity markers using a sandwich cultured human hepatocyte model leveraging primary human hepatocytes from pediatric and adult donors. This will involve measuring proteomic and metabolomic biomarkers in drug-treated hepatocyte lots from infants, children, adolescents, and adults to understand age-specific mechanisms of DILI. The study will also analyze proteomics data of extracellular vesicles and untargeted metabolomics of media to identify molecular changes and drug-protein adducts. These data will be integrated into mathematical models using quantitative systems toxicology (QST) models to predict pathways and mechanisms of DILI. Aim 2 will measure the effects of acute and prolonged drug exposure using liver microphysiological system (MPS) using standardized liver tissue chip. By analyzing proteins and metabolites released in the media from these systems, the study will assess immediate and long-term impacts on liver function and toxicity in both pediatric and adult livers. Overall, utilizing advanced methodologies such as proteomics, metabolomics, and complex in vitro models, including MPS, the research will assess DILI across various pediatric age groups. Integrating these findings into mathematical models will provide a deeper understanding of DILI mechanisms and potentially identify specific biomarkers relevant to children. Ultimately, when validated in clinic, pediatric DILI candidate biomarkers will enhance drug safety profiles, inform better prescribing practices, and support personalized treatment approaches in children.
