Lineage recording reveals hijacked hepatic progenitor states as a common origin of HCC and ICC

Highlights

CERAMIC enables continuous and high-capacity lineage tracing of liver tumor initiation

Fatty liver-associated hepatocytes acquire regenerative and premalignant cell states before malignant transformation

Lineage reconstruction identifies Hep_Bi-zonal cells as the cellular origin of liver tumor initiation

Transcriptional and regulatory programs distinguish tumor-fated hepatocytes from failed-to-transform lineages

Peroxisomal metabolism is required for progenitor-state formation and liver tumor initiation

Spatial remodeling identifies a macrophage niche associated with tumor-fated hepatocytes

Dual ontogenies and functional specialization of lipid-associated macrophages shape the tumor-fated hepatocyte niche

Fatty liver disease predisposes to primary liver cancer, yet the lineage routes and niche mechanisms that select rare tumor-fated hepatocytes remain unclear. Here we developed CERAMIC, a high-capacity CRISPR-Cas9 lineage recorder that co-recovers editing scars and transcriptomes from single cells, and applied it to an AKT/NRAS-driven model of MASLD-associated liver tumor initiation. Longitudinal lineage, single-cell and spatial analyses revealed a hierarchical trajectory in which bipotential bi-zonal hepatocytes (Hep_Bi-zonal), rather than pericentral-like hepatocytes (Hep_CVlike), generated regenerative and neoplastic hepatocyte progenitor states that progressed toward both hepatocellular carcinoma and intrahepatic cholangiocarcinoma lineages. Tumor-fated cells preferentially expanded along a remodeled midlobular-periportal axis and depended on ACOX1-mediated peroxisomal beta-oxidation to withstand lipotoxic and oxidative stress. Spatial and lineage analyses further identified a sequential lipid-associated macrophage niche, in which monocyte-derived LAMs engaged tumor-fated hepatocytes through an LGALS9-P4HB axis, and P4HB inhibition suppressed tumor expansion. These findings define liver tumor initiation as a lineage-restricted process licensed by peroxisomal metabolic adaptation and macrophage-derived niche signals.