Université de Paris, Innovative Therapies in Haemostasis, INSERM, France (N.G., A.B., E.R., S.I., S.L., A. Cras, N.N., J.R., P.G., D.M.S.).
Hematology Department and Biosurgical Research Lab (Carpentier Foundation) (N.G., A.B., E.R., S.L., N.N., J.R., P.G., D.M.S.), AH-HP, Georges Pompidou European Hospital, France.
Arterioscler Thromb Vasc Biol. 2021 Jan;41(1):415-429. doi: 10.1161/ATVBAHA.120.314287. Epub 2020 Nov 5.
The study's aim was to analyze the capacity of human valve interstitial cells (VICs) to participate in aortic valve angiogenesis. Approach and Results: VICs were isolated from human aortic valves obtained after surgery for calcific aortic valve disease and from normal aortic valves unsuitable for grafting (control VICs). We examined VIC in vitro and in vivo potential to differentiate in endothelial and perivascular lineages. VIC paracrine effect was also examined on human endothelial colony-forming cells. A pathological VIC (VIC) mesenchymal-like phenotype was confirmed by CD90/CD73/CD44 expression and multipotent-like differentiation ability. When VIC were cocultured with endothelial colony-forming cells, they formed microvessels by differentiating into perivascular cells both in vivo and in vitro. VIC and control VIC conditioned media were compared using serial ELISA regarding quantification of endothelial and angiogenic factors. Higher expression of VEGF (vascular endothelial growth factor)-A was observed at the protein level in VIC-conditioned media and confirmed at the mRNA level in VIC compared with control VIC. Conditioned media from VIC induced in vitro a significant increase in endothelial colony-forming cell proliferation, migration, and sprouting compared with conditioned media from control VIC. These effects were inhibited by blocking VEGF-A with blocking antibody or siRNA approach, confirming VIC involvement in angiogenesis by a VEGF-A dependent mechanism.
We provide here the first proof of an angiogenic potential of human VICs isolated from patients with calcific aortic valve disease. These results point to a novel function of VIC in valve vascularization during calcific aortic valve disease, with a perivascular differentiation ability and a VEGF-A paracrine effect. Targeting perivascular differentiation and VEGF-A to slow calcific aortic valve disease progression warrants further investigation.
本研究旨在分析人心脏瓣膜间质细胞（VICs）参与主动脉瓣血管生成的能力。
我们从因钙化性主动脉瓣疾病而手术切除的人主动脉瓣以及不适合移植的正常主动脉瓣（对照 VICs）中分离出 VICs。我们在体外和体内检查了 VIC 向血管内皮细胞和血管周细胞谱系分化的潜能。还研究了 VIC 的旁分泌作用对人内皮集落形成细胞的影响。通过 CD90/CD73/CD44 表达和多能样分化能力证实了病理性 VIC（VIC）间充质样表型。当 VIC 与内皮集落形成细胞共培养时，它们在体内和体外均分化为血管周细胞形成微血管。通过连续 ELISA 比较 VIC 和对照 VIC 条件培养基，以定量检测内皮和血管生成因子。VIC 条件培养基中 VEGF-A（血管内皮生长因子-A）的蛋白水平表达较高，VIC 中的 mRNA 水平也证实了这一点，与对照 VIC 相比。与对照 VIC 的条件培养基相比，VIC 的条件培养基在体外诱导内皮集落形成细胞增殖、迁移和出芽显著增加。这些作用被 VEGF-A 阻断抗体或 siRNA 方法阻断，证实了 VIC 通过 VEGF-A 依赖机制参与血管生成。
我们在此首次提供了从钙化性主动脉瓣疾病患者中分离出的人 VIC 具有血管生成潜力的证据。这些结果表明 VIC 在钙化性主动脉瓣疾病期间瓣膜血管化中具有新的功能，具有血管周细胞分化能力和 VEGF-A 的旁分泌作用。针对血管周细胞分化和 VEGF-A 以减缓钙化性主动脉瓣疾病进展值得进一步研究。
