肝切除术后肝衰竭（PHLF）导致接受肝切除术的患者预后不良，其中肝血管重建发挥着关键作用。然而，肝血管重建的调节因子仍不清楚。本研究旨在探讨肝血管重建的调控机制，并鉴定预测肝切除患者 PHLF 的生物标志物。结合 Alb-Cre-CRISPR/ 技术，在腺相关病毒 8（AAV8）中筛选与肝血管重建相关的候选基因。 Cas9小鼠接受了部分肝切除术。使用内皮前体输注和用于分期肝切除术（ALPPS）模型的相关肝脏分区和门静脉结扎来评估候选基因的生物活性。在 ALPPS 患者的活组织检查中检测到候选物的水平。利用回顾性数据筛选PHLF的危险因素。肝细胞中Gata3的下调和Ramp2的上调促进了肝窦内皮细胞（LSEC）的增殖和肝血运重建。色素上皮衍生因子（PEDF）和血管内皮生长因子A（VEGFA）在调节肝切除后内皮前体从骨髓的迁移和新血窦的形成中发挥相反的作用。 Gata3 限制患者来源的肝类器官中的内皮细胞功能，但 Gata3 抑制剂可消除这种功能。此外，Gata3 的过度表达导致 ALPPS 小鼠死亡率较高，PEDF 中和抗体可改善这种情况。 ALPPS患者中Gata3/Ramp2与PEDF/VEGFA的表达呈负相关。构建了结合血清PEDF/VEGF指数（SPVI）等多种因素的列线图模型，可以有效预测PHLF的风险。肝细胞中Gata3和Ramp2的平衡通过将PEDF转移到VEGFA来调节LSEC的增殖和肝血运重建，从而促进肝血管重建。为预防和治疗PHLF提供了潜在的靶点。在这项研究中，我们揭示了一种新的机制，即肝细胞中Gata3和Ramp2的平衡通过在肝切除或ALLPS诱导的肝再生过程中将PEDF转变为VEGFA来调节肝血管重建。我们还发现血清 PEDF/VEGFA 指数 (SPVI) 是接受肝切除术的患者肝切除术后肝衰竭 (PHLF) 的潜在预测因子。这项研究将有助于更好地了解肝细胞如何促进肝脏再生以及预防和治疗 PHLF 的新靶点。版权所有 © 2023。由 Elsevier B.V. 出版。

Post-hepatectomy liver failure (PHLF) leads to poor prognosis in patients receiving hepatectomy, where hepatic-vascular reconstitution plays a critical role. However, the regulators of hepatic-vascular reconstitution remain unclear. This study aimed to investigate the regulatory mechanisms of hepatic-vascular reconstitution and identify biomarkers predicting PHLF in patients underwent hepatectomy.Candidate genes that were associated with hepatic vascular reconstitution were screened in adeno-associated virus 8 (AAV8) combining Alb-Cre-CRISPR/Cas9 mice underwent partial hepatectomy. The biological activities of candidate genes were estimated using endothelial precursors transfusion and an associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) model. The level of candidates was detected in biopsies from ALPPS patients. Risk factors of PHLF were also screened using retrospective data.The downregulation of Gata3 and upregulation of Ramp2 in hepatocytes promoted the proliferation of liver sinusoidal endothelial cells (LSECs) and hepatic revascularization. Pigment epithelium-derived factor (PEDF) and vascular endothelial growth factor A (VEGFA) played opposite roles in regulating the migration of endothelial precursors from bone marrow and the formation of new sinusoids after hepatectomy. Gata3 restricted endothelial cell function in patient-derived hepatic-organoids, which was abrogated by Gata3 inhibitor. Moreover, overexpression of Gata3 led to higher mortality in ALPPS mice, which was improved by PEDF-neutralizing antibody. The expression of Gata3/Ramp2 and PEDF/VEGFA tended to have negative correlation in ALPPS patients. A nomogram model incorporating multiple factors such as serum PEDF/VEGF index (SPVI) was constructed and could efficiently predict the risk of PHLF.The balance of Gata3 and Ramp2 in hepatocytes regulated the proliferation of LSECs and hepatic revascularization via shifting PEDF to VEGFA, which provided potential targets for the prevention and treatment of PHLF.In this study, we revealed a novel mechanism that the balance of Gata3 and Ramp2 in hepatocyte regulated hepatic vascular reconstitution via shifting PEDF to VEGFA during hepatectomy- or ALLPS-induced liver regeneration. We also identified serum PEDF/VEGFA index (SPVI) as a potential predictor for post-hepatectomy liver failure (PHLF) in patients who underwent hepatectomy. This study will provide a better understanding on how hepatocytes contribute to liver regeneration and new targets for the prevention and treatment of PHLF.Copyright © 2023. Published by Elsevier B.V.