Endothelial Hyaluronan Synthase 3 Augments Postischemic Arteriogenesis Through CD44/eNOS Signaling
Endothelial Hyaluronan Synthase 3 Augments Postischemic Arteriogenesis Through CD44/eNOS Signaling
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DOI：
10.1161/atvbaha.121.315478
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发表时间：
2021-10-01
影响因子：
8.7
通讯作者：
Grandoch, Maria
中科院分区：
文献类型：
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作者：
Schneckmann, Rebekka;Suvorava, Tatsiana;Grandoch, Maria
目的：动脉形成的主要驱动因素是内皮糖萼感知的剪应力升高，从而促进动脉向外重构。透明质酸是内皮糖萼的重要组成部分，由质膜上的3种HAS同工酶（透明质酸合成酶1-3）合成。进一步考虑到HAS3对平滑肌细胞和免疫细胞功能的重要性，我们旨在评估其在侧支动脉生长中的作用。方法与结果：雄性has3缺陷（Has3-KO）小鼠后肢缺血。用激光多普勒血流显像监测血流灌注，用体内血流介导的舒张测量评估内皮功能。采用高分辨率磁共振血管造影监测侧支血管重构。腹腔注射CD44中和抗体（克隆KM201），分析体内透明质酸信号传导。后肢缺血后，Has3-KO小鼠的动脉生成反应降低，侧支重塑减少，灌注恢复受损。虽然缺血后白细胞浸润未受影响，但血流介导的扩张减弱表明内皮细胞功能受损。事实上，在Has3-KO大腿肌肉中，内皮AKT（蛋白激酶B）依赖的eNOS（内皮一氧化氮合酶）在Ser1177位点的磷酸化显著降低。内皮特异性Has3-KO小鼠模拟了在全球has3缺乏症中观察到的后肢缺血诱导的灌注恢复受损表型。在机制上，选择性阻断CD44的透明质酸结合位点可减少血流介导的扩张，从而表明通过CD44传递透明质酸信号是潜在的信号通路。结论：综上所述，通过透明质酸/ cd44介导的eNOS Ser1177磷酸化刺激，HAS3参与后肢缺血的动脉发生。因此，可以设想增加内皮细胞HAS3或CD44的策略来增强病理条件下的血管化。
Objective: The dominant driver of arteriogenesis is elevated shear stress sensed by the endothelial glycocalyx thereby promoting arterial outward remodeling. Hyaluronan, a critical component of the endothelial glycocalyx, is synthesized by 3 HAS isoenzymes (hyaluronan synthases 1-3) at the plasma membrane. Considering further the importance of HAS3 for smooth muscle cell and immune cell functions we aimed to evaluate its role in collateral artery growth. Approach and Results: Male Has3-deficient (Has3-KO) mice were subjected to hindlimb ischemia. Blood perfusion was monitored by laser Doppler perfusion imaging and endothelial function was assessed by measurement of flow-mediated dilation in vivo. Collateral remodeling was monitored by high resolution magnetic resonance angiography. A neutralizing antibody against CD44 (clone KM201) was injected intraperitoneally to analyze hyaluronan signaling in vivo. After hindlimb ischemia, Has3-KO mice showed a reduced arteriogenic response with decreased collateral remodeling and impaired perfusion recovery. While postischemic leukocyte infiltration was unaffected, a diminished flow-mediated dilation pointed towards an impaired endothelial cell function. Indeed, endothelial AKT (protein kinase B)-dependent eNOS (endothelial nitric oxide synthase) phosphorylation at Ser1177 was substantially reduced in Has3-KO thigh muscles. Endothelial-specific Has3-KO mice mimicked the hindlimb ischemia-induced phenotype of impaired perfusion recovery as observed in global Has3-deficiency. Mechanistically, blocking selectively the hyaluronan binding site of CD44 reduced flow-mediated dilation, thereby suggesting hyaluronan signaling through CD44 as the underlying signaling pathway. Conclusions: In summary, HAS3 contributes to arteriogenesis in hindlimb ischemia by hyaluronan/CD44-mediated stimulation of eNOS phosphorylation at Ser1177. Thus, strategies augmenting endothelial HAS3 or CD44 could be envisioned to enhance vascularization under pathological conditions.