ANXA2+ migratory hepatocytes identified as key players in human liver regeneration

Acute liver failure (ALF) represents a life-threatening condition marked by rapid deterioration of liver function, often necessitating urgent liver transplantation. In the United States, acetaminophen (APAP) overdose remains the most common cause of drug-induced ALF, accounting for nearly half of all cases. Given the scarcity of viable donor livers—worsened by the prevalence of steatotic liver disease—understanding and promoting the liver's intrinsic regenerative capacity has become an urgent priority in hepatology.

Liver regeneration following injury is known to rely primarily on hepatocyte proliferation and, to a lesser extent, trans-differentiation from cholangiocytes. However, in cases of ALF, particularly those reaching the stage requiring transplantation, histological analysis often reveals insufficient or ineffective tissue repair, suggesting gaps in our understanding of the regenerative process.

This study builds upon recent advances in single-cell and spatial transcriptomics, integrating multimodal approaches across human and mouse models to elucidate cellular dynamics during liver regeneration. The goal was to identify specific mechanisms that could be targeted to promote effective wound healing and limit necrotic progression in the liver.

Multimodal Approach to Liver Regeneration

The research team employed a robust and integrative methodology that included: (1) Single-nucleus RNA sequencing (snRNA-seq); (2) Spatial transcriptomics; (3) Multiplex single-molecule fluorescence in situ hybridization (MsmFISH); (4) Four-dimensional intravital microscopy; (5) Lineage tracing in reporter mice. This combination allowed for a high-resolution spatiotemporal map of liver cell behavior in both human APAP-ALF tissue and APAP-injured mouse models.

Key Discovery: ANXA2+ Migratory Hepatocytes

A major finding of the study was the identification of a distinct subpopulation of hepatocytes expressing annexin A2 (ANXA2), located along the border of necrotic regions. These cells exhibited: (1) Ruffled membranes and lamellipodia (hallmarks of migratory morphology); (2) Expression profiles consistent with both portal and central hepatocyte gene modules; (3) Coordinated interactions with hepatic stellate cells (HSCs) and immune cells.

Using intravital imaging and lineage tracing in mice, the researchers showed that: (1) Hepatocyte necrosis peaked around 30 hours post-APAP injury; (2) Hepatocyte proliferation peaked at 72 hours; (3) Migration of ANXA2+ hepatocytes occurred between 36 and 42 hours. Further, a majority of glutamine synthetase-positive hepatocytes near necrotic zones were BrdU-negative, supporting the notion that these cells arrived via migration rather than proliferation.

Lineage tracing also confirmed that ANXA2+ cells originated from pre-existing hepatocytes, not from cholangiocytes or hepatic progenitor cells. This finding further emphasizes the liver's ability to re-purpose differentiated hepatocytes in response to acute damage.

Functional Role of ANXA2

To determine the functional relevance of ANXA2, the authors performed knockdown experiments: (1) In vitro scratch assays using Huh7 cells and primary mouse hepatocytes showed reduced wound closure with ANXA2 knockdown; (2) In vivo knockdown of ANXA2 using AAV8-shRNA led to impaired wound closure without affecting hepatocyte proliferation. These results suggest that ANXA2 is a critical mediator of hepatocyte migration and wound healing, functioning independently of cell division.

Clinical and Therapeutic Potential

This research highlights the potential of harnessing endogenous liver repair mechanisms to treat ALF. Therapeutic strategies aimed at enhancing ANXA2 expression or function could: (1) accelerate liver regeneration; (2) reduce the need for liver transplantation; (3) improve outcomes in patients presenting beyond the metabolic phase of APAP toxicity. Furthermore, targeting hepatocyte migration, rather than solely proliferation, adds a new dimension to regenerative medicine approaches in hepatology.

In conclusion, the identification of ANXA2+ migratory hepatocytes represents a paradigm shift in our understanding of liver regeneration. By elucidating a novel mechanism of wound closure in the liver, this study opens new therapeutic avenues for acute liver failure, with broader implications for chronic liver diseases and regenerative medicine.

 

See the article:

Nguyen K, Ding WX, Ni H-M. Multimodal decoding of human liver regeneration uncovers novel ANXA2+ migratory hepatocytes for wound healing. eGastroenterology 2025;3:e100205. doi:10.1136/ egastro-2025-100205

 

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