Department of Pathology, University of Washington, Seattle, WA, USA.
Pathology and Laboratory Medicine Service (S-113-Lab), VA Puget Sound Health Care System, Seattle, WA, USA.
Angiogenesis. 2019 Aug;22(3):421-431. doi: 10.1007/s10456-019-09667-z. Epub 2019 Apr 9.
Progression of atherosclerotic plaques into life-threatening lesions is associated with angiogenesis which contributes to intraplaque hemorrhages and plaque instability. The lack of adequate models for the study of human plaque-induced angiogenesis has limited progress in this field. We describe here a novel ex vivo model which fills this gap. Plaques obtained from 15 patients who underwent endarterectomy procedures were co-cultured in collagen gels with rat aorta rings which served as read-out of human plaque angiogenic activity. The majority of plaque fragments markedly stimulated angiogenic sprouting from the aortic rings while concurrently promoting the outgrowth of resident macrophages from the aortic adventitia. This stimulatory activity correlated with the presence of intraplaque macrophages. Proteomic analysis of plaque secretomes revealed heterogeneity of macrophage-stimulatory cytokine and angiogenic factor production by different plaques. VEGF was identified in some of the plaque secretomes. Antibody-mediated blockade of VEGF had significant but transient inhibitory effect on angiogenesis, which suggested redundancy of plaque-derived angiogenic stimuli. Pharmacologic ablation of adventitial macrophages permanently impaired the angiogenic response of aortic rings to plaque stimuli. Our results show that human plaque-induced angiogenesis can be reproduced ex vivo using rat aortic rings as read-out of plaque angiogenic activity. This model can be used to identify key cellular and molecular mechanisms responsible for the neovascularization of human plaques.
动脉粥样硬化斑块进展为危及生命的病变与血管生成有关，血管生成有助于斑块内出血和斑块不稳定。缺乏用于研究人类斑块诱导的血管生成的充分模型，限制了该领域的进展。我们在这里描述了一种新的离体模型来填补这一空白。从接受内膜切除术的 15 名患者中获得的斑块与大鼠主动脉环在胶原凝胶中共培养，作为人类斑块血管生成活性的读出器。大多数斑块碎片明显刺激了主动脉环的血管生成发芽，同时促进了主动脉外膜中固有巨噬细胞的生长。这种刺激活性与斑块内巨噬细胞的存在相关。斑块分泌组的蛋白质组学分析显示不同斑块产生的巨噬细胞刺激细胞因子和血管生成因子存在异质性。一些斑块分泌组中存在 VEGF。VEGF 的抗体介导阻断对血管生成有显著但短暂的抑制作用，这表明斑块来源的血管生成刺激物具有冗余性。外膜巨噬细胞的药理学消融永久损害了主动脉环对斑块刺激的血管生成反应。我们的结果表明，使用大鼠主动脉环作为斑块血管生成活性的读出器，可以在离体条件下重现人类斑块诱导的血管生成。该模型可用于鉴定负责人类斑块新生血管形成的关键细胞和分子机制。
