OBJECTIVE Although often studied independently, little is known about how aortic valve endothelial cells and valve interstitial cells interact collaborate to maintain tissue homeostasis or drive valve calcific pathogenesis. Inflammatory signaling is a recognized initiator of valve calcification, but the cell-type-specific downstream mechanisms have not been elucidated. In this study, we test how inflammatory signaling via NFκB (nuclear factor κ-light-chain enhancer of activated B cells) activity coordinates unique and shared mechanisms of valve endothelial cells and valve interstitial cells differentiation during calcific progression. Approach and Results: Activated NFκB was present throughout the calcific aortic valve disease (CAVD) process in both endothelial and interstitial cell populations in an established mouse model of hypercholesterolemia-induced CAVD and in human CAVD. NFκB activity induces endothelial to mesenchymal transformation in 3-dimensional cultured aortic valve endothelial cells and subsequent osteogenic calcification of transformed cells. Similarly, 3-dimensional cultured valve interstitial cells calcified via NFκB-mediated osteogenic differentiation. NFκB-mediated endothelial to mesenchymal transformation was directly demonstrated in vivo during CAVD via genetic lineage tracking. Genetic deletion of NFκB in either whole valves or valve endothelium only was sufficient to prevent valve-specific molecular and cellular mechanisms of CAVD in vivo despite the persistence of a CAVD inducing environment. CONCLUSIONS Our results identify NFκB signaling as an essential molecular regulator for both valve endothelial and interstitial participation in CAVD pathogenesis. Direct demonstration of valve endothelial cell endothelial to mesenchymal transformation transmigration in vivo during CAVD highlights a new cellular population for further investigation in CAVD morbidity. The efficacy of valve-specific NFκB modulation in inhibiting hypercholesterolemic CAVD suggests potential benefits of multicell type integrated investigation for biological therapeutic development and evaluation for CAVD.

中文翻译：

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NFκB（激活的B细胞的核因子κ-轻链增强剂）活性调节特定于细胞类型和特定于上下文的主动脉瓣钙化敏感性。

目的尽管经常进行独立研究，但对主动脉瓣内皮细胞和瓣膜间质细胞如何相互作用以维持组织稳态或驱动瓣膜钙化发病机制了解甚少。炎症信号是公认的瓣膜钙化的起因，但​​尚未阐明细胞类型特异性的下游机制。在这项研究中，我们测试了钙化进程中通过NFκB（活化的B细胞的核因子κ-轻链增强剂）活性的炎症信号如何协调独特的和共有的瓣膜内皮细胞和瓣膜间质细胞分化的机制。方法和结果：在已建立的高胆固醇血症诱发的CAVD小鼠模型和人CAVD的内皮细胞和间质细胞群中，活化的NFκB存在于整个钙化主动脉瓣疾病（CAVD）过程中。NFκB活性诱导3维培养的主动脉瓣内皮细胞的内皮向间充质转化，以及随后的转化细胞成骨钙化。同样，通过NFκB介导的成骨分化，钙化的3维瓣膜间质细胞也被钙化。在CAVD期间，通过遗传谱系跟踪直接证明了NFκB介导的内皮细胞向间充质转化。尽管持续存在CAVD诱导环境，但在整个瓣膜或瓣膜内皮中仅通过基因缺失NFκB就足以预防CAVD体内瓣膜特异性分子和细胞机制。结论我们的结果确定了NFκB信号传导是瓣膜内皮和间质参与CAVD发病机制的必要分子调节剂。在CAVD期间体内瓣膜内皮细胞向间充质转化内皮细胞迁移的直接展示突出了一个新的细胞群体，需要进一步研究CAVD的发病率。阀特异性NFκB调节抑制高胆固醇血症CAVD的功效表明，多细胞类型整合研究对于CAVD的生物治疗发展和评估具有潜在的益处。结论我们的结果确定了NFκB信号传导是瓣膜内皮和间质参与CAVD发病机制的必要分子调节剂。在CAVD期间体内瓣膜内皮细胞向间充质转化内皮细胞迁移的直接展示突出了一个新的细胞群体，需要进一步研究CAVD的发病率。阀特异性NFκB调节抑制高胆固醇血症CAVD的功效表明，多细胞类型整合研究对于CAVD的生物治疗发展和评估具有潜在的益处。结论我们的结果确定了NFκB信号传导是瓣膜内皮和间质参与CAVD发病机制的必要分子调节剂。在CAVD期间体内瓣膜内皮细胞向间充质转化内皮细胞迁移的直接展示突出了一个新的细胞群体，需要进一步研究CAVD的发病率。阀特异性NFκB调节抑制高胆固醇血症CAVD的功效表明，多细胞类型整合研究对于CAVD的生物治疗发展和评估具有潜在的益处。在CAVD期间体内瓣膜内皮细胞向间充质转化内皮细胞迁移的直接展示突出了一个新的细胞群体，需要进一步研究CAVD的发病率。阀特异性NFκB调节抑制高胆固醇血症CAVD的功效表明，多细胞类型整合研究对于CAVD的生物治疗发展和评估具有潜在的益处。在CAVD期间体内瓣膜内皮细胞向间充质转化内皮细胞迁移的直接展示突出了一个新的细胞群体，需要进一步研究CAVD的发病率。阀特异性NFκB调节抑制高胆固醇血症CAVD的功效表明，多细胞类型整合研究对于CAVD的生物治疗发展和评估具有潜在的益处。