Table_1_sVEGFR1 Is Enriched in Hepatic Vein Blood—Evidence for a Provisional Hepatic Factor Candidate?.docx

Background: Pulmonary arteriovenous malformations (PAVMs) are common sequelae of palliated univentricular congenital heart disease, yet their pathogenesis remain poorly defined. In this preliminary study, we used paired patient blood samples to identify potential hepatic factor candidates enriched in hepatic vein blood.
Methods: Paired venous blood samples were collected from the hepatic vein (HV) and superior vena cava (SVC) from children 0 to 10 years with univentricular and biventricular congenital heart disease (n = 40). We used three independent protein analyses to identify proteomic differences between HV and SVC blood. Subsequently, we investigated the relevance of our quantified protein differences with human lung microvascular endothelial assays.
Results: Two independent protein arrays (semi-quantitative immunoblot and quantitative array) identified that soluble vascular endothelial growth factor receptor 1 (sVEGFR1) is significantly elevated in HV serum compared to SVC serum. Using ELISA, we confirmed the previous findings that sVEGFR1 is enriched in HV serum (n = 24, p < 0.0001). Finally, we studied the quantified HV and SVC serum levels of sVEGFR1 in vitro. HV levels of sVEGFR1 decreased tip cell selection (p = 0.0482) and tube formation (fewer tubes [p = 0.0246], shorter tube length [p = 0.0300]) in vitro compared to SVC levels of sVEGFR1.
Conclusions: Based on a small heterogenous cohort, sVEGFR1 is elevated in HV serum compared to paired SVC samples, and the mean sVEGFR1 concentrations in these two systemic veins cause pulmonary endothelial phenotypic differences in vitro. Further research is needed to determine whether sVEGFR1 has a direct role in pulmonary microvascular remodeling and PAVMs in patients with palliated univentricular congenital heart disease.

研究背景：肺动静脉畸形（Pulmonary arteriovenous malformations, PAVMs）是接受姑息治疗的单心室先天性心脏病患者的常见后遗症，但其发病机制仍未明确。本初步研究采用配对患者血液样本，旨在鉴定肝静脉血液中富集的潜在肝脏因子候选物。
研究方法：本研究纳入0至10岁的单心室及双心室先天性心脏病患儿共40例（n=40），从其肝静脉（hepatic vein, HV）与上腔静脉（superior vena cava, SVC）采集配对静脉血样本。我们采用三种独立的蛋白质分析手段，鉴定HV与SVC血液的蛋白质组学差异。随后，我们探究了定量得到的蛋白质差异与人类肺微血管内皮细胞实验的相关性。
研究结果：两种独立的蛋白质芯片（半定量免疫印迹与定量蛋白芯片）分析显示，可溶性血管内皮生长因子受体1（soluble vascular endothelial growth factor receptor 1, sVEGFR1）在HV血清中的水平显著高于SVC血清。通过酶联免疫吸附试验（ELISA），我们验证了此前的发现：HV血清中sVEGFR1富集（n=24，p<0.0001）。最后，我们在体外环境下对HV与SVC血清中定量得到的sVEGFR1水平进行了研究。与SVC来源的sVEGFR1相比，HV来源的sVEGFR1在体外可降低尖端细胞选择能力（p=0.0482）并抑制管腔形成（管腔数量减少[p=0.0246]，管腔长度缩短[p=0.0300]）。
研究结论：基于小型异质性队列研究，与配对SVC样本相比，HV血清中sVEGFR1水平升高，且这两条体静脉的sVEGFR1平均浓度可在体外引发肺内皮细胞表型差异。仍需开展进一步研究，以明确sVEGFR1在接受姑息治疗的单心室先天性心脏病患者的肺微血管重构与肺动静脉畸形中是否发挥直接作用。