MASH and Human Liver Spheroids
Background and Aims
Metabolic dysfunction-associated steatohepatitis (MASH) is a progressive severe disease characterized by hepatic lipid accumulation, inflammation, and fibrosis, which lacks any approved drug therapy. Novel approaches to identify therapeutic candidates that predict clinical responses are needed. Human pre-clinical models, including organoids/spheroids, are powerful models for drug testing and discovery. Pre-clinical models combining fast, robust, high-throughput compatible, and predictive outputs are still a limiting factor in identifying novel clinical therapies.
We aimed to perform a comparison of the safety and efficacy of existing clinical candidates for MASH in a high-throughput manner using a human-derived 3D liver spheroid model and multiplex readouts.
Method
We used a previously developed human-derived 3D liver spheroid model in 96-well plates containing primary human hepatocytes, Kupffer cells, endothelial cells, and stellate cells from healthy donors. Incubation with a MASH-inducing cocktail of the 3D liver model for 10 days led to the recapitulation of the main hallmarks of MASH, including steatosis, inflammation, and fibrosis.
We screened simultaneously the safety and efficacy of 20 current clinical candidates in a concentration-dependent manner. We selected secreted lactate dehydrogenase (LDH), intracellular triglycerides (TGs), and secreted pro-collagen 1 (PC-1) as surrogate markers of cytotoxicity, steatosis, and fibrosis. LDH was measured on days 3, 5, 7, and 10; TGs on day 10; and PC-1 on days 7 and 10.
Results
The screen of 20 clinical candidates comprised different modalities, including small molecules, antibodies, and antisense oligos (ASOs) targeting different mechanisms of action. The majority of compounds did not show signs of toxicity at lower doses, but some led to high LDH release at higher doses. We generated a matrix of anti-steatotic versus anti-fibrotic effects to plot the comparison of all tested clinical candidates. Examples of successful translational effects are the FASN inhibitor TVB-2640 (Sagimet), showing a significant reduction of triglycerides, and the alpha/beta1 integrin inhibitor (PLN-1474), demonstrating a strong reduction of PC-1 secreted levels.
Conclusion
We show for the first time a screening platform that combines safety and MASH efficacy readouts. We show the translational capacity of this human in vitro 3D platform by recapitulating the clinical effects of several drug candidates.
Download the poster about the human liver spheroids
