Human stem cell-derived hepatic cells recapitulate polygenic risk in MASLD development and highlight CD36 as a key mediator

Several genetic variants have been identified that impact outcomes in patients with metabolic dysfunction-associated steatohepatitis (MASH), but cellular platforms to identify molecular targets for treatment remain scarce. This study aims to employ a polygenic risk score for hepatic fat content (PRS-HFC) in a human-based in vitro model to identify potential druggable targets related to a genetic risk for MASH. Ninety-two human skin-derived precursor (hSKP) cell lines were genotyped and classified according to their PRS-HFC. Per risk category, five cell lines were selected and differentiated to hepatic progenitor-like cells (hSKP-HPCs) before being exposed to MASH-related triggers or control medium. Lipid accumulation, fatty acid uptake, generation of oxidative stress, secretion of inflammatory mediators, and alterations of the transcriptome were assessed. Potential drug targets were modulated using siRNA. hSKP-HPCs displayed a positive correlation between lipid accumulation and PRS-HFC, both in the control and ‘MASH’ condition. hSKP-HPCs with high PRS-HFC exhibited increased secretion of interleukin 6 and enhanced fatty acid uptake. Bulk RNA barcoding and sequencing revealed distinct gene expression profiles depending on genetic predisposition and predicted increased CD36-signalling as potential upstream regulator thereof. Silencing of CD36 effectively reduced lipid accumulation in cultures with high PRS-HFC and attenuated expression of genes related to de novo lipogenesis, lipolysis and inflammation. hSKP-HPCs recapitulate the effect of the PRS-HFC on lipid accumulation and inflammation . CD36 appeared to at least partially mediate these effects, and its silencing further substantiated its role in conferring high genetic risk for MASH, highlighting CD36 as a potential therapeutic target for genetically-driven MASH. Abbreviations ANOVA analysis of variance B2M beta-2-microglobulin BMI body mass index BMP4 bone morphogenetic protein 4 BRBseq bulk RNA barcoding and sequencing BSA bovine serum albumin CCL C-C motif chemokine ligand CD36 cluster of differentiation 36 cDNA copy DNA ChREBP carbohydrate-response element-binding protein DEGs differentially expressed genes DGAT2 diacylglycerol O-acyltransferase 2 DMSO dimethyl sulfoxide DMEM Dulbecco's Modified Eagle Medium ELISA enzyme-linked immunosorbent assay FASN fatty acid synthase FATP fatty acid transport protein FBS foetal bovine serum FDA Food and Drug Administration FGF fibroblast growth factor GCKR glucokinase regulator gDNA genomic DNA GPX7 glutathione peroxidase 7 HBSS Hanks' Balanced Salt Solution HCC hepatocellular carcinoma HGF hepatocyte growth factor hSKP human skin-derived precursor hSKP-HPC human skin-derived precursor hepatic progenitor-like cells IL interleukin INSIG2 insulin induced gene 2 IPA Ingenuity Pathway Analysis ITS insulin-transferrin‑selenium LPS lipopolysaccharide MASH metabolic dysfunction-associated steatohepatitis MASLD metabolic dysfunction-associated steatotic liver disease MBOAT7 membrane bound O-acyltransferase domain-containing 7 NQO1 nicotinamide adenine dinucleotide (phosphate) quinone dehydrogenase 1 PCR polymerase chain reaction PNPLA3 patatin-like phospholipase domain-containing protein 3 PRS-HFC polygenic risk score for hepatic fat content ROS reactive oxygen species SAA serum amyloid A SCAP SREBP cleavage-activating protein SCD stearoyl-coenzyme A desaturase SD standard deviation siRNA small interfering RNA SNP single-nucleotide polymorphism SOD2 superoxide dismutase 2 SREBP sterol regulatory element binding protein SSO sodium sulfosuccinimidyl oleate STAT3 signal transducer and activator of transcription 3 TBS/T Tris-buffered saline with 0.1% (v/v) Tween® 20 TM6SF2 transmembrane 6 superfamily member 2 TNFα tumour necrosis factor alpha UBC ubiquitin C URA upstream regulator analysis VLDL very low-density lipoprotein.