Shinohara Tsutomu, Moonen Jan-Renier, Chun Yoon Hong, Lee-Yow Yannick C, Okamura Kenichi, Szafron Jason M, Kaplan Jordan, Cao Aiqin, Wang Lingli, Guntur Divya, Taylor Shalina, Isobe Sarasa, Dong Melody, Yang Weiguang, Guo Katherine, Franco Benjamin D, Pacharinsak Cholawat, Pisani Laura J, Saitoh Shinji, Mitani Yoshihide, Marsden Alison L, Engreitz Jesse M, Körbelin Jakob, Rabinovitch Marlene
Department of Pediatrics (T.S., J.-R.M., Y.H.C., J.M.S., J. Kaplan, A.C., L.W., D.G., S.T., S.I., M.D., W.Y., A.L.M., M.R.).
Stanford Cardiovascular Institute (T.S., J.-R.M., Y.H.C., K.O., J.M.S., J. Kaplan, A.C., L.W., S.T., S.I., M.D., A.L.M., J.M.E., M.R.), Stanford University School of Medicine, CA.
Arterioscler Thromb Vasc Biol. 2025 Feb;45(2):218-237. doi: 10.1161/ATVBAHA.124.321092. Epub 2024 Dec 26.
Computational modeling indicated that pathological high shear stress (HSS; 100 dyn/cm) is generated in pulmonary arteries (PAs; 100-500 µm) in congenital heart defects causing PA hypertension (PAH) and in idiopathic PAH with occlusive vascular remodeling. Endothelial-to-mesenchymal transition (EndMT) is a feature of PAH. We hypothesize that HSS induces EndMT, contributing to the initiation and progression of PAH.
We used the Ibidi perfusion system to determine whether HSS applied to human PA endothelial cells (ECs) induces EndMT when compared with physiological laminar shear stress (15 dyn/cm). The mechanism was investigated and targeted to prevent PAH in a mouse with HSS induced by an aortocaval shunt.
EndMT, a feature of PAH not previously attributed to HSS, was observed. HSS did not alter the induction of transcription factors KLF (Krüppel-like factor) 2/4, but an ERG (ETS-family transcription factor) was reduced, as were histone H3 lysine 27 acetylation enhancer-promoter peaks containing ERG motifs. Consequently, there was reduced interaction between ERG and KLF2/4, a feature important in tethering KLF and the chromatin remodeling complex to DNA. In PA ECs under laminar shear stress, reducing ERG by siRNA caused EndMT associated with decreased BMPR2 (bone morphogenetic protein receptor 2), CDH5 (cadherin 5), and PECAM1 (platelet and EC adhesion molecule 1) and increased SNAI1/2 (Snail/Slug) and ACTA2 (smooth muscle α2 actin). In PA ECs under HSS, transfection of ERG prevented EndMT. HSS was then induced in mice by an aortocaval shunt, causing progressive PAH over 8 weeks. An adeno-associated viral vector (AAV2-ESGHGYF) was used to replenish ERG selectively in PA ECs. Elevated PA pressure, EndMT, and vascular remodeling (muscularization of peripheral arteries) in the aortocaval shunt mice were markedly reduced by ERG delivery.
Pathological HSS reduced lung EC ERG, resulting in EndMT and PAH. Agents that upregulate ERG could reverse HSS-mediated PAH and occlusive vascular remodeling resulting from high flow or narrowed PAs.
计算模型表明，在导致肺动脉高压（PAH）的先天性心脏缺陷以及伴有闭塞性血管重塑的特发性PAH患者的肺动脉（PA；直径100 - 500 µm）中会产生病理性高剪切应力（HSS；100达因/平方厘米）。内皮 - 间充质转化（EndMT）是PAH的一个特征。我们假设HSS诱导EndMT，促进PAH的发生和发展。
我们使用Ibidi灌注系统来确定与生理层流剪切应力（15达因/平方厘米）相比，施加于人类PA内皮细胞（ECs）的HSS是否会诱导EndMT。对其机制进行了研究，并旨在预防由主动脉腔静脉分流诱导HSS的小鼠发生PAH。
观察到EndMT这一PAH的特征，此前未将其归因于HSS。HSS并未改变转录因子KLF（Krüppel样因子）2/4的诱导，但ERG（ETS家族转录因子）减少，含有ERG基序的组蛋白H3赖氨酸27乙酰化增强子 - 启动子峰也减少。因此，ERG与KLF2/4之间的相互作用减少，这是将KLF和染色质重塑复合物与DNA拴系在一起的一个重要特征。在层流剪切应力下的PA ECs中，通过小干扰RNA降低ERG会导致EndMT，同时伴有骨形态发生蛋白受体2（BMPR2）、钙黏蛋白5（CDH5）和血小板与内皮细胞黏附分子1（PECAM1）减少，以及蜗牛家族转录抑制因子1/2（SNAI1/2）和平滑肌α2肌动蛋白（ACTA2）增加。在HSS下的PA ECs中，转染ERG可预防EndMT。然后通过主动脉腔静脉分流在小鼠中诱导HSS，导致在8周内逐渐发生PAH。使用腺相关病毒载体（AAV2 - ESGHGYF）在PA ECs中选择性补充ERG。通过递送ERG，主动脉腔静脉分流小鼠升高的PA压力、EndMT和血管重塑（外周动脉肌化）明显减少。
病理性HSS降低肺EC中的ERG，导致EndMT和PAH。上调ERG的药物可逆转HSS介导的PAH以及由高流量或狭窄PA导致的闭塞性血管重塑。
