Spatial single-cell transcriptomics of MASLD-affected liver biopsy samples.

Metabolic dysfunction-associated steatotic liver disease (MASLD) liver biopsies were analyzed using spatial single-cell transcriptomics (CosMx Spatial Molecular Imager, NanoString) to elucidate molecular alterations associated with hepatic fibrosis morphology, as quantified by the artificial intelligence-based FibroNest algorithm (PharmaNest). This analysis identified distinct morphological fiber phenotypes (FibroPCs), among which FibroPC4, characterized by reticular fiber structures, was associated with a hepatic stellate cell (HSC) phenotype that promotes hepatocellular carcinoma (HCC). This HCC-promoting HSC phenotype was driven by insulin-like growth factor-binding protein 7 (IGFBP-7) secreted from senescent periportal endothelial cells. Spatial single-cell transcriptomic profiling using the NanoString CosMx Spatial Molecular Imager was performed on formalin-fixed paraffin-embedded (FFPE) liver biopsy specimens obtained from four patients with metabolic dysfunction-associated steatotic liver disease (MASLD) and fibrosis stage F2. Two biopsy sections from different patients were mounted on each slide for analysis.