Modeling the pathogenic mechanisms of metabolic dysfunction-associated steatotic liver disease (MASLD) progression using specialized human multicellular 3D organotypic models
Year: 2025
Type: dissertation
Abstract: Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) is the most common chronic liver disease, affecting approximately 38% of the global adult population. It encompasses a spectrum of conditions, ranging from simple steatosis (Metabolic Dysfunction-Associated Steatotic Liver, MASL) to more advanced stages, such as Metabolic Dysfunction-Associated Steatohepatitis (MASH), marked by inflammation and fibrosis, which may progress to cirrhosis and hepatocellular carcinoma. Unequivocal diagnosis still relies primarily on invasive liver biopsy. Treatment focuses on lifestyle modifications, which are challenging to sustain and less effective in advanced stages. Understanding the multifactorial mechanisms driving MASLD progression, particularly chronic inflammation, is essential for developing improved diagnostic tools and effective therapies. This study aimed to improve and enhance a recently developed 3D multicellular hepatic model by Baptista et al (2024)., which was shown to recapitulate MASLD mechanisms. Improvements included optimizing culture conditions using serum alternatives, refining biomarkers of MASL/MASH progression, and assessing the model’s ability to replicate in vivo liver architecture and injury response. It also investigated the role of Cytochrome P450 oxidoreductase (CPR, POR encoded), a redox partner in both CYP and non-CYP enzyme-mediated reactions, whose direct impact on the MASLD pathogenesis had not been confirmed. By using both 2D and 3D HepaRG models, culture conditions could be optimized, resulting in the regularization of cellular lipid homeostasis, and thereby increasing the applicability of this cell model for MASLD studies. Analysis of multiple potential biomarkers confirmed the model’s suitability for tracking MASL/MASH progression and elucidating key disease mechanisms. CPR’s role was explored via POR gene knockdown, confirming its involvement in lipid accumulation and mitochondrial stress, hallmarks of MASLD progression. This study successfully optimized the 3D hepatic models developed by Baptista et al. (2024), validated their relevance for MASLD research, and provided novel insights into CPR’s role in disease progression.
Author: Ganço, Sara
Language: English
Subfield: Hepatology
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W7115976607