We developed a novel, patient-relevant three-dimensional (3D) liver spheroid model composed of primary human hepatocytes and non-parenchymal cells (hepatic stellate cells, Kupffer cells, and liver sinusoidal endothelial cells). This model replicates key histopathological features of MASH, including steatosis, inflammation, and fibrosis, closely matching patient biopsy data. The spheroids spontaneously develop fibrosis, which worsens with free fatty acid treatment. Liver endothelial cells (LSECs) were found to drive fibrosis by releasing tissue inhibitor of metalloproteinases 1 (TIMP1), which is upregulated under lipotoxic stress and enriched in fibrotic liver zones. Knockdown of TIMP1 reduced collagen accumulation, confirming its role in fibrosis regulation.
To advance therapeutic discovery, the model was combined with high-content imaging, multi-omics profiling, and chemogenomic screening. This revealed promising therapeutic targets such as muscarinic M1 receptor (CHRM1) activation and TRPM8 inhibition. Mechanistic studies showed that CHRM1 inhibits TRPM8 via phospholipase C activation, leading to strong anti-fibrotic effects.
Additionally, we developed a long-term 3D spheroid model for screening conjugated siRNAs for specificity, stability, off target effects and toxicity. These spheroids maintained viability and asialoglycoprotein receptor expression for five weeks, enabling sustained, non-toxic siRNA uptake and durable gene silencing. This human-relevant platform provides a crucial bridge between traditional in vitro systems and animal models for siRNA therapeutic development.
We built a strong network of over 20 partners, including pharmaceutical companies and academic researchers focused on MASH. The 3D MASH model has been successfully integrated into its service portfolio, with multiple industry requests for testing candidate molecules’ anti-fibrotic and anti-steatotic effects. These results validate the model’s commercialization potential for early preclinical testing. Chemogenomic screens have also identified novel drug targets, with lead-like molecules in development and evaluation through genetic tools like KO mice. Intellectual property from these efforts will be carefully managed. Notably, HepaPredict entered a licensing agreement with Shanghai Hepo Biotechnology Ltd, granting exclusive rights to deploy the 3D MASH technology in China, demonstrating the model’s maturity and commercial attractiveness for future investment.
