Abstract：
Post-hepatectomy liver failure (PHLF) leads to poor prognosis in patients undergoing hepatectomy, with hepatic vascular reconstitution playing a critical role. However, the regulators of hepatic vascular reconstitution remain unclear. In this study, we aimed to investigate the regulatory mechanisms of hepatic vascular reconstitution and identify biomarkers predicting PHLF in patients undergoing hepatectomy.
Candidate genes that were associated with hepatic vascular reconstitution were screened using adeno-associated virus vectors in Alb-Cre-CRISPR/Cas9 mice subjected to partial hepatectomy. The biological activities of candidate genes were estimated using endothelial precursor transfusion and associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) models. The level of candidates was detected in biopsies from patients undergoing ALPPS. Risk factors for PHLF were also screened using retrospective data.
Downregulation of Gata3 and upregulation of Ramp2 in hepatocytes promoted the proliferation of liver sinusoidal endothelial cells 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 a Gata3 inhibitor. Moreover, overexpression of Gata3 led to higher mortality in ALPPS mice, which was improved by a PEDF-neutralizing antibody. The expression of Gata3/RAMP and PEDF/VEGFA tended to have a negative correlation in patients undergoing ALPPS. A nomogram incorporating multiple factors, such as serum PEDF/VEGF index, was constructed and could efficiently predict the risk of PHLF.
The balance of Gata3 and Ramp2 in hepatocytes regulates the proliferation of liver sinusoidal endothelial cells and hepatic revascularization via changes in the expression of PEDF and VEGFA, revealing potential targets for the prevention and treatment of PHLF.
In this study, we show that the balance of Gata3 and Ramp2 in hepatocytes regulates hepatic vascular reconstitution by promoting a shift from pigment epithelium-derived factor (PEDF) to vascular endothelial growth factor A (VEGFA) expression during hepatectomy- or ALLPS (associating liver partition and portal vein ligation for staged hepatectomy)-induced liver regeneration. We also identified serum PEDF/VEGFA index as a potential predictor of post-hepatectomy liver failure in patients who underwent hepatectomy. This study improves our understanding of how hepatocytes contribute to liver regeneration and provides new targets for the prevention and treatment of post-hepatectomy liver failure.
Candidate genes that were associated with hepatic vascular reconstitution were screened using adeno-associated virus vectors in Alb-Cre-CRISPR/Cas9 mice subjected to partial hepatectomy. The biological activities of candidate genes were estimated using endothelial precursor transfusion and associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) models. The level of candidates was detected in biopsies from patients undergoing ALPPS. Risk factors for PHLF were also screened using retrospective data.
Downregulation of Gata3 and upregulation of Ramp2 in hepatocytes promoted the proliferation of liver sinusoidal endothelial cells 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 a Gata3 inhibitor. Moreover, overexpression of Gata3 led to higher mortality in ALPPS mice, which was improved by a PEDF-neutralizing antibody. The expression of Gata3/RAMP and PEDF/VEGFA tended to have a negative correlation in patients undergoing ALPPS. A nomogram incorporating multiple factors, such as serum PEDF/VEGF index, was constructed and could efficiently predict the risk of PHLF.
The balance of Gata3 and Ramp2 in hepatocytes regulates the proliferation of liver sinusoidal endothelial cells and hepatic revascularization via changes in the expression of PEDF and VEGFA, revealing potential targets for the prevention and treatment of PHLF.
In this study, we show that the balance of Gata3 and Ramp2 in hepatocytes regulates hepatic vascular reconstitution by promoting a shift from pigment epithelium-derived factor (PEDF) to vascular endothelial growth factor A (VEGFA) expression during hepatectomy- or ALLPS (associating liver partition and portal vein ligation for staged hepatectomy)-induced liver regeneration. We also identified serum PEDF/VEGFA index as a potential predictor of post-hepatectomy liver failure in patients who underwent hepatectomy. This study improves our understanding of how hepatocytes contribute to liver regeneration and provides new targets for the prevention and treatment of post-hepatectomy liver failure.
摘要：
目的：肝切除术后肝功能衰竭（PHLF）导致肝切除术患者预后不良，其中肝血管重建起着关键作用。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，并确定预测肝切除术患者PHLF的生物标志物。
方法：在腺相关病毒8（AAV8）联合Alb-Cre-CRISPR/Cas9小鼠中筛选与肝血管重建相关的候选基因。使用内皮前体输血和相关的肝分区和门静脉结扎用于分期肝切除术（ALPPS）模型来评估候选基因的生物学活性。在ALPPS患者的活检中检测到候选水平。同时使用回顾性资料筛选PHLF的危险因素。
结果：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）在调节骨髓内皮前体的迁移和肝切除术后新窦的形成中起相反的作用。Gata3限制了患者来源的肝类器官中的内皮细胞功能，已被Gata3抑制剂废除。此外，Gata3的过度表达导致ALPPS小鼠的死亡率更高，通过PEDF中和抗体改善。ALPPS患者Gata3/Ramp2与PEDF/VEGFA的表达呈负相关。构建了血清PEDF/VEGF指数（SPVI）等多因素的列线图模型，可以有效预测PHLF的风险。
结论：肝细胞中Gata3和Ramp2的平衡通过将PEDF转变为VEGFA来调节LSECs的增殖和肝血运重建，这为PHLF的预防和治疗提供了潜在的目标。
■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。我们还确定了血清PEDF/VEGFA指数（SPVI）作为肝切除术后肝衰竭（PHLF）患者的潜在预测因子。这项研究将更好地了解肝细胞如何促进肝脏再生和预防和治疗PHLF的新目标。
方法：在腺相关病毒8（AAV8）联合Alb-Cre-CRISPR/Cas9小鼠中筛选与肝血管重建相关的候选基因。使用内皮前体输血和相关的肝分区和门静脉结扎用于分期肝切除术（ALPPS）模型来评估候选基因的生物学活性。在ALPPS患者的活检中检测到候选水平。同时使用回顾性资料筛选PHLF的危险因素。
结果：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）在调节骨髓内皮前体的迁移和肝切除术后新窦的形成中起相反的作用。Gata3限制了患者来源的肝类器官中的内皮细胞功能，已被Gata3抑制剂废除。此外，Gata3的过度表达导致ALPPS小鼠的死亡率更高，通过PEDF中和抗体改善。ALPPS患者Gata3/Ramp2与PEDF/VEGFA的表达呈负相关。构建了血清PEDF/VEGF指数（SPVI）等多因素的列线图模型，可以有效预测PHLF的风险。
结论：肝细胞中Gata3和Ramp2的平衡通过将PEDF转变为VEGFA来调节LSECs的增殖和肝血运重建，这为PHLF的预防和治疗提供了潜在的目标。
■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。我们还确定了血清PEDF/VEGFA指数（SPVI）作为肝切除术后肝衰竭（PHLF）患者的潜在预测因子。这项研究将更好地了解肝细胞如何促进肝脏再生和预防和治疗PHLF的新目标。
