The liver has a unique ability to regenerate^1,2; however, in the setting of acute liver failure (ALF), this regenerative capacity is often overwhelmed, leaving emergency liver transplantation as the only curative option^3,4,5. Here, to advance understanding of human liver regeneration, we use paired single-nucleus RNA sequencing combined with spatial profiling of healthy and ALF explant human livers to generate a single-cell, pan-lineage atlas of human liver regeneration. We uncover a novel ANXA2⁺ migratory hepatocyte subpopulation, which emerges during human liver regeneration, and a corollary subpopulation in a mouse model of acetaminophen (APAP)-induced liver regeneration. Interrogation of necrotic wound closure and hepatocyte proliferation across multiple timepoints following APAP-induced liver injury in mice demonstrates that wound closure precedes hepatocyte proliferation. Four-dimensional intravital imaging of APAP-induced mouse liver injury identifies motile hepatocytes at the edge of the necrotic area, enabling collective migration of the hepatocyte sheet to effect wound closure. Depletion of hepatocyte ANXA2 reduces hepatocyte growth factor-induced human and mouse hepatocyte migration in vitro, and abrogates necrotic wound closure following APAP-induced mouse liver injury. Together, our work dissects unanticipated aspects of liver regeneration, demonstrating an uncoupling of wound closure and hepatocyte proliferation and uncovering a novel migratory hepatocyte subpopulation that mediates wound closure following liver injury. Therapies designed to promote rapid reconstitution of normal hepatic microarchitecture and reparation of the gut–liver barrier may advance new areas of therapeutic discovery in regenerative medicine.

肝脏具有独特的再生能力^1,2；然而，在急性肝衰竭 (ALF) 的情况下，这种再生能力常常被压垮，使得紧急肝移植成为唯一的治疗选择^3,4,5。在这里，为了加深对人类肝脏再生的理解，我们使用配对单核 RNA 测序结合健康和 ALF 外植体人类肝脏的空间分析，生成人类肝脏再生的单细胞、全谱系图谱。我们发现了人类肝脏再生过程中出现的新型 ANXA2 ⁺迁移肝细胞亚群，以及对乙酰氨基酚 (APAP) 诱导的肝再生小鼠模型中的推论亚群。对 APAP 诱导的小鼠肝损伤后多个时间点的坏死伤口闭合和肝细胞增殖的研究表明，伤口闭合先于肝细胞增殖。 APAP 诱导的小鼠肝损伤的四维活体成像可识别坏死区域边缘的活动肝细胞，使肝细胞片集体迁移以实现伤口闭合。肝细胞 ANXA2 的消耗可减少肝细胞生长因子诱导的人和小鼠肝细胞体外迁移，并消除 APAP 诱导的小鼠肝损伤后的坏死性伤口闭合。我们的工作共同剖析了肝再生的意想不到的方面，证明了伤口闭合和肝细胞增殖的解耦，并揭示了一种新的迁移肝细胞亚群，可介导肝损伤后的伤口闭合。旨在促进正常肝脏微结构快速重建和肠肝屏障修复的疗法可能会推动再生医学治疗发现的新领域。