Single-Cell RNA Sequencing Uncovers Novel Cell States in Regenerating Liver Tissue
DOI:
https://doi.org/10.5281/zenodo.22687944Keywords:
single-cell RNA sequencing, liver regeneration, partial hepatectomy, hepatocyte heterogeneity, pseudotime, macrophage polarisationAbstract
The mammalian liver is unusual among solid organs in being able to restore its full mass within days of substantial
tissue loss, yet the cellular logic of that recovery is still described mostly in terms of bulk proliferation indices rather
than the identities of the cells doing the work. We performed droplet-based single-cell RNA sequencing on liver
tissue from C57BL/6 mice at six time points after two-thirds partial hepatectomy (0, 24, 48, 72, 96 and 168 hours),
profiling 58,412 cells that passed quality control across eighteen animals. Unsupervised clustering resolved the
expected hepatocyte, endothelial, Kupffer cell, stellate cell and cholangiocyte compartments, but also identified
two transient cell states not present in sham-operated controls: a periportal 'regenerative hepatocyte' population
marked by co-expression of Afp, Spp1 and cell-cycle genes at 48-72 hours, and a distinct Ly6c-high, Vegfa-high
macrophage state that peaked at 24-48 hours and preceded the appearance of the regenerative hepatocyte
cluster in pseudotime. Ligand-receptor analysis linked this macrophage state to hepatocyte proliferation via a
Vegfa-Flt1/Kdr axis and an Spp1-Cd44 axis, and depletion of Kupffer cells with clodronate liposomes prior to
hepatectomy delayed the emergence of the regenerative hepatocyte cluster and reduced Ki67-positive hepatocyte
fraction at 72 hours by roughly half. Together, these data reposition regenerative hepatocytes and Vegfa-high
macrophages as coupled, time-restricted cell states whose sequential appearance forms part of the coordination
logic of liver regeneration, rather than as regeneration being explained by hepatocyte proliferation in isolation.
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