Dissecting compounded hepatocyte injury in a model of MASLD progression from human induced pluripotent stem cells

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a complex, multifactorial condition influenced by genetic predisposition, inter-organ signaling, and immune cell infiltration. The difficulty of modeling the complex nature of the disease makes it increasingly challenging to identify treatments that intercept its progression. Here, we show the suitability of cumulative injury modeling toward steatohepatitis from hepatocyte-like cells (HLCs) generated from human induced pluripotent stem cells (iPSCs). These include insults such as free fatty acid excess, soluble extra-hepatic signaling, pro-inflammatory immune cell co-culture and genomic modifiers (PNPLA3-I148M variant). We harness this model to elucidate the molecular and functional consequences of hepatocellular injury progression and identify suitable points of therapeutic interception. Both healthy and PNPLA3-I148M iPSC-donor derived HLCs developed high-level generic steatosis response, yet the PNPLA3-I148M line was more susceptible to mitochondrial injury by the adipokine resistin and the myokine myostatin. Co-culture with peripheral blood mononuclear cells (PBMCs) compounded the previous multi-hit injury. Further, transcriptomic data revealed inflammatory HLC reprogramming and loss of genomically encoded mitochondrial metabolism along injury progression. We demonstrate a nuanced spectral metabolic injury model, mirroring clinical MASLD progression, suitable for testing stage and donor-specific disease interceptions.