Secreted Factors From Proinflammatory Macrophages Promote an Osteoblast-Like Phenotype in Valvular Interstitial Cells.
Secreted Factors From Proinflammatory Macrophages Promote an Osteoblast-Like Phenotype in Valvular Interstitial Cells.
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DOI：
10.1161/atvbaha.120.315261
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发表时间：
2020-11
期刊：
影响因子：
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通讯作者：
Anseth KS
中科院分区：
文献类型：
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作者：
Grim JC;Aguado BA;Vogt BJ;Batan D;Andrichik CL;Schroeder ME;Gonzalez-Rodriguez A;Yavitt FM;Weiss RM;Anseth KS
在主动脉瓣狭窄（AVS）进展期间，驻留的瓣膜间质细胞（VIC）活化为肌成纤维细胞，这进一步促进纤维化，甚至分化为成骨细胞样细胞，可导致瓣膜组织钙化。炎症是AVS的标志，因此我们旨在确定M1巨噬细胞分泌的促炎细胞因子，这些细胞因子引起短暂的维克表型，导致瓣膜组织钙化。我们设计了水凝胶生物材料作为瓣膜细胞外基质模拟物，使VIC能够在其静止成纤维细胞或活化肌成纤维细胞表型中培养，以响应局部基质硬度。当用THP-1衍生的M1巨噬细胞的条件培养基处理维克成纤维细胞和肌成纤维细胞时，我们观察到α-平滑肌肌动蛋白（α SMA）表达的显著降低、应力纤维形成减少和增殖增加，表明具有强效抗纤维化作用。我们进一步确定了M1培养基中与观察到的抗纤维化作用相关的两种细胞因子为肿瘤坏死因子α（TNF-α）和白细胞介素1 β（IL-1 β）。在M1条件培养基中培养7天后，VIC开始分化为成骨细胞样细胞，如通过增加RUNX2和骨桥蛋白的表达所测量的。我们还确定并验证了IL-6作为观察到的促成骨作用的关键介质。M1条件培养基中的促炎细胞因子，特别是TNF-α、IL-1 β和IL-6，抑制VIC中的肌成纤维细胞反应并促进其成骨分化。总之，我们的工作表明，炎症M1巨噬细胞可能驱动肌成纤维细胞到成骨中间维克表型，这可能介导从纤维化到钙化的转换在AVS进展。
Resident valvular interstitial cells (VICs) activate to myofibroblasts during aortic valve stenosis (AVS) progression, which further promotes fibrosis or even differentiate into osteoblast-like cells that can lead to calcification of valve tissue. Inflammation is a hallmark of AVS, so we aimed to determine pro-inflammatory cytokines secreted from M1 macrophages that give rise to a transient VIC phenotype that leads to calcification of valve tissue. We designed hydrogel biomaterials as valve extracellular matrix mimics enabling the culture of VICs in either their quiescent fibroblast or activated myofibroblast phenotype in response to the local matrix stiffness. When VIC fibroblasts and myofibroblasts were treated with conditioned media from THP-1-derived M1 macrophages, we observed robust reduction of α-smooth muscle actin (αSMA) expression, reduced stress fiber formation, and increased proliferation, suggesting a potent anti-fibrotic effect. We further identified that two cytokines in M1 media attributed to the observed anti-fibrotic effects were tumor necrosis factor alpha (TNF-α) and interleukin 1 beta (IL-1β). After 7 days of culture in M1 conditioned media, VICs began differentiating into osteoblast-like cells, as measured by increased expression of RUNX2 and osteopontin. We also identified and validated IL-6 as a critical mediator of the observed pro-osteogenic effect. Pro-inflammatory cytokines in M1 conditioned media, notably TNF-α, IL-1β, and IL-6, inhibit the myofibroblast response in VICs and promote their osteogenic differentiation. Together, our work suggests inflammatory M1 macrophages may drive a myofibroblast-to-osteogenic intermediate VIC phenotype, which may mediate the switch from fibrosis to calcification during AVS progression.