Valve endothelial-interstitial interactions drive emergent complex calcific lesion formation in vitro.
Valve endothelial-interstitial interactions drive emergent complex calcific lesion formation in vitro.
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瓣膜内皮-间质相互作用驱动体外紧急复杂钙化病变形成。
DOI：
10.1016/j.biomaterials.2021.120669
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发表时间：
2021-03
期刊：
影响因子：
14
通讯作者：
中科院分区：
文献类型：
--
作者：
钙化性主动脉瓣疾病（CAVD）是一种主动调节的退行性疾病过程。临床病变表现出明显的3D复杂性，在目前的体外系统中没有表现出来。我们在这里提出了一个独特的机械应力三维培养系统，重演瓣膜间质细胞（维克）诱导的基质钙化，通过成肌纤维细胞活化和成骨细胞分化。我们测试的假设，即瓣膜内皮（VEC）间质协作相互作用调节的风险和复杂性的钙化发病机制内的机械应力和促炎环境。将猪主动脉瓣内皮细胞和间质细胞（VEC和维克）单独或以共培养配置接种在机械约束的胶原水凝胶中。当在成骨培养基（OGM）中培养时，在7天内形成凸起的3D VIC-filled病变，并且令人惊讶地通过内皮共培养而加剧。我们确定了一个空间协调的pro-endochondrial与pro-osteogenic信号程序内病变。VEC经历了内皮-间充质转化（EndMT）并聚集在病变中心。该3D体外CAVD系统的分子和细胞特征的空间复杂性与人类患病主动脉瓣组织学一致。SNAI 1在VEC和内皮下直接维克中高表达，证实了人CAVD病变。Sox 9与Runx 2表达在已发展病变内的空间分布（Sox 9病变周围与Runx 2主要在病变内）反映了它们在严重钙化的人主动脉瓣中的表达。最后，我们证明了该平台通过BAY 11-7082阻断经典NFκB通路来筛选潜在药物治疗的适用性。我们的研究结果表明，VEC通过EndMT和旁分泌信号积极诱导维克病理性重构和钙化。这种机械约束的培养平台使得能够询问由这种细胞反馈回路支持的加速的细胞介导的基质重塑行为。这种复杂的3D模型系统对人类CAVD机制的高保真度支持其用于测试瓣膜中细胞间通讯的机制及其药理学控制。
Calcific aortic valve disease (CAVD) is an actively regulated degenerative disease process. Clinical lesions exhibit marked 3D complexity not represented in current in vitro systems. We here present a unique mechanically stressed 3D culture system that recapitulates valve interstitial cell (VIC) induced matrix calcification through myofibroblastic activation and osteoblastic differentiation. We test the hypothesis that valve endothelial (VEC) – interstitial collaborative interactions modulate the risk and complexity of calcific pathogenesis within mechanically stressed and pro-inflammatory environments. Porcine aortic valve endothelial and interstitial cells (VEC and VIC) were seeded in a mechanically constrained collagen hydrogels alone or in co-culture configurations. Raised 3D VIC-filled lesions formed within 7 days when cultured in osteogenic media (OGM), and surprisingly exacerbated by endothelial coculture. We identified a spatially coordinated pro-endochondral vs. pro-osteogenic signaling program within the lesion. VEC underwent Endothelial-to-Mesenchymal Transformation (EndMT) and populated the lesion center. The spatial complexity of molecular and cellular signatures of this 3D in vitro CAVD system were consistent with human diseased aortic valve histology. SNAI1 was highly expressed in the VEC and sub-endothelial direct VIC corroborates with human CAVD lesions. Spatial distribution of Sox9 vs. Runx2 expression within the developed lesions (Sox9 peri-lesion vs. Runx2 predominantly within lesions) mirrored their expression in heavily calcified human aortic valves. Finally, we demonstrate the applicability of this platform for screening potential pharmacologic therapies through blocking the canonical NFκB pathway via BAY 11–7082. Our results establish that VEC actively induce VIC pathological remodeling and calcification via EndMT and paracrine signaling. This mechanically constrained culture platform enables the interrogation of accelerated cell-mediated matrix remodeling behavior underpinned by this cellular feedback circuit. The high fidelity of this complex 3D model system to human CAVD mechanisms supports its use to test mechanisms of intercellular communication in valves and their pharmacological control.
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影响因子：
14
作者：
Farrar, Emily J.;Pramil, Varsha;Richards, Jennifer M.;Mosher, Christopher Z.;Butcher, Jonathan T.
通讯作者：
Butcher, Jonathan T.
影响因子：
2.9
作者：
Gilda JE;Gomes AV
通讯作者：
Gomes AV
蜗牛1在心脏肌纤维细胞中的作用对于低氧损伤后心脏纤维化很重要。
影响因子：
3.7
作者：
Biswas H;Longmore GD
通讯作者：
Longmore GD
DOI：
10.1161/atvbaha.115.306091
发表时间：
2016-02
期刊：
Arteriosclerosis, thrombosis, and vascular biology
影响因子：
--
作者：
Huk DJ;Austin BF;Horne TE;Hinton RB;Ray WC;Heistad DD;Lincoln J
通讯作者：
Lincoln J
影响因子：
5
作者：
Anstine, Lindsey J.;Bobba, Chris;Lincoln, Joy
通讯作者：
Lincoln, Joy