From the Cardiovascular Translational Research, Navarrabiomed, Complejo Hospitalario de Navarra (CHN), Universidad Pública de Navarra (UPNA), IdiSNA, Pamplona, Spain (L.M., J.I., V. Arrieta, A.G.-P., A.F.-C., A.N., V. Álvarez, A.G., R.S., N.L.-A.).
Laboratory of Atherothrombosis, Program of Cardiovascular Diseases, CIMA Universidad de Navarra, IdiSNA, Pamplona, Spain (C.R., J.O., J.A.R.).
Arterioscler Thromb Vasc Biol. 2020 May;40(5):1370-1382. doi: 10.1161/ATVBAHA.120.314143. Epub 2020 Mar 19.
Aortic valve (AV) calcification plays an important role in the progression of aortic stenosis (AS). MMP-10 (matrix metalloproteinase-10 or stromelysin-2) is involved in vascular calcification in atherosclerosis. We hypothesize that MMP-10 may play a pathophysiological role in calcific AS. Approach and Results: Blood samples (n=112 AS and n=349 controls) and AVs (n=88) from patients undergoing valve replacement were analyzed. Circulating MMP-10 was higher in patients with AS compared with controls (<0.001) and correlated with TNFα (tumor necrosis factor α; r=0.451; <0.0001). MMP-10 was detected by immunochemistry in AVs from patients with AS colocalized with aortic valve interstitial cells markers α-SMA (α-smooth muscle actin) and vimentin and with calcification markers Runx2 (Runt-related transcription factor 2) and SRY (sex-determining region Y)-box 9. MMP-10 expression in AVs was further confirmed by RT-qPCR and western blot. Ex vivo, MMP-10 was elevated in the conditioned media of AVs from patients with AS and associated with interleukin-1β (r=0.5045, <0.001) and BMP (bone morphogenetic protein)-2 (r=0.5003, <0.01). In vitro, recombinant human MMP-10 induced the overexpression of inflammatory, fibrotic, and osteogenic markers (interleukin-1β, α-SMA, vimentin, collagen, BMP-4, Sox9, OPN [osteopontin], BMP-9, and Smad 1/5/8; <0.05) and cell mineralization in aortic valve interstitial cells isolated from human AVs, in a mechanism involving Akt (protein kinase B) phosphorylation. These effects were prevented by TIMP-1 (tissue inhibitor of metalloproteinases type 1), a physiological MMP inhibitor, or specifically by an anti-MMP-10 antibody.
MMP-10, which is overexpressed in aortic valve from patients with AS, seems to play a central role in calcification in AS through Akt phosphorylation. MMP-10 could be a new therapeutic target for delaying the progression of aortic valve calcification in AS.
主动脉瓣（AV）钙化在主动脉瓣狭窄（AS）的进展中起着重要作用。基质金属蛋白酶 10（MMP-10 或基质溶解素 2）参与动脉粥样硬化中的血管钙化。我们假设 MMP-10 可能在钙化性 AS 中发挥病理生理作用。方法和结果：分析了 88 例接受瓣膜置换术的患者的血液样本（AS 患者 112 例，对照组 349 例）和 AV 样本。与对照组相比，AS 患者的循环 MMP-10 水平更高（<0.001），并与 TNFα（肿瘤坏死因子α；r=0.451；<0.0001）相关。免疫化学检测到 AS 患者的 AV 中 MMP-10 与主动脉瓣间质细胞标志物 α-SMA（α-平滑肌肌动蛋白）和波形蛋白共定位，并与钙化标志物 Runx2（Runt 相关转录因子 2）和 SRY（性别决定区 Y）-box 9 共定位。AV 中的 MMP-10 表达进一步通过 RT-qPCR 和 Western blot 得到证实。在体外，AS 患者的 AV 条件培养基中 MMP-10 升高，并与白细胞介素 1β（r=0.5045，<0.001）和 BMP（骨形态发生蛋白）-2（r=0.5003，<0.01）相关。在体外，重组人 MMP-10 诱导从人 AV 分离的主动脉瓣间质细胞中炎症、纤维化和成骨标志物（白细胞介素 1β、α-SMA、波形蛋白、胶原、BMP-4、Sox9、OPN[骨桥蛋白]、BMP-9 和 Smad 1/5/8）的过度表达和细胞矿化，在这种机制中涉及 Akt（蛋白激酶 B）磷酸化。这些作用可通过 TIMP-1（基质金属蛋白酶抑制剂 1 型），一种生理 MMP 抑制剂，或特异性抗 MMP-10 抗体来预防。结论：在 AS 患者的主动脉瓣中过度表达的 MMP-10 似乎通过 Akt 磷酸化在 AS 中的钙化中发挥核心作用。MMP-10 可能成为延迟 AS 中主动脉瓣钙化进展的新治疗靶点。
