Adamts5-/- Mice Exhibit Altered Aggrecan Proteolytic Profiles That Correlate With Ascending Aortic Anomalies
Adamts5-/- Mice Exhibit Altered Aggrecan Proteolytic Profiles That Correlate With Ascending Aortic Anomalies
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DOI：
10.1161/atvbaha.119.313077
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发表时间：
2019-10-01
影响因子：
8.7
通讯作者：
Kern, Christine B.
中科院分区：
文献类型：
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作者：
Dupuis, Loren E.;Nelson, E. Lockett;Kern, Christine B.
目的：研究ADAMTS（一种具有血小板反应蛋白型基序的去整合素样和金属蛋白酶结构域）5缺陷和二叶式主动脉瓣的小鼠模型中主动脉壁发育期间聚集蛋白聚糖（Acan）裂解的需求。方法：将检查蛋白聚糖细胞外基质重塑改变的小鼠升主动脉壁发育异常。识别ADAMTS切割的Acan片段的新表位抗体将用于研究主动脉壁发育中Acan周转的机制要求。结果如下：Adamts 5（-/-）; Smad 2（+/-）小鼠显示出较高的主动脉畸形发生率（n=17/17）;具有二叶主动脉瓣的Adamts 5（-/-）; Smad 2（+/-）小鼠（7/17）显示出比具有三叶主动脉瓣的Adamts 5（-/-）; Smad 2（+/-）小鼠更多的异常。单突变型Adamts 5（-/-）小鼠也显示出高的主动脉畸形发生率（n=19/19），与野生型（n=1/11）相比。主动脉异常与Acan蓄积相关，这在Adamts 5（-/-）小鼠的弹性细胞生成开始时是明显的。与WT相比，识别Acan切割片段neo-FREEE、neo-GLGS和neo-SSELE中的初始氨基酸的新表位抗体在Adamts 5（-/-）Adamts中增加。相反，识别高度消化的Acan核心片段的neo-TEGE在Adamts 5（-/-）小鼠中减少。然而，在TEGE（373）向下箭头（374）ALGSV位点中含有突变（使其不可裂解）的小鼠具有低的主动脉异常发生率（n=2/4）。在主动脉外膜中观察到Acan neo-DIPEN和neo-FFGVG片段;在中层中Acan neo-FFGVG异常增加，并与Adamts 5（-/-）动脉中的平滑肌细胞丢失重叠。结论：ADAMTS 5 Acan在发育过程中的断裂与升主动脉畸形相关。这些数据表明ADAMTS 5 Acan切割机制可能对正常主动脉壁发育至关重要。
Objective: Investigate the requirement of Aggrecan (Acan) cleavage during aortic wall development in a murine model with ADAMTS (a disintegrin-like and metalloprotease domain with thrombospondin-type motifs) 5 deficiency and bicuspid aortic valves. Approach: Mice with altered extracellular matrix remodeling of proteoglycans will be examined for anomalies in ascending aortic wall development. Neo-epitope antibodies that recognize ADAMTS cleaved Acan fragments will be used to investigate the mechanistic requirement of Acan turnover, in aortic wall development. Results: Adamts5(-/-);Smad2(+/-) mice exhibited a high penetrance of aortic anomalies (n=17/17); Adamts5(-/-);Smad2(+/-) mice with bicuspid aortic valves (7/17) showed a higher number of anomalies than Adamts5(-/-);Smad2(+/-) mice with tricuspid aortic valves. Single mutant Adamts5(-/-) mice also displayed a high penetrance of aortic anomalies (n=19/19) compared with wild type (n=1/11). Aortic anomalies correlated with Acan accumulation that was apparent at the onset of elastogenesis in Adamts5(-/-) mice. Neo-epitope antibodies that recognize the initial amino acids in the Acan cleaved fragments neo-FREEE, neo-GLGS, and neo-SSELE were increased in the Adamts5(-/-) aortas compared with WT. Conversely, neo-TEGE, which recognizes highly digested Acan core fragments, was reduced in Adamts5(-/-) mice. However, mice containing a mutation in the TEGE(373)down arrow(374)ALGSV site, rendering it noncleavable, had low penetrance of aortic anomalies (n=2/4). Acan neo-DIPEN and neo-FFGVG fragments were observed in the aortic adventitia; Acan neo-FFGVG was increased abnormally in the medial layer and overlapped with smooth muscle cell loss in Adamts5(-/-) aortas. Conclusions: Disruption of ADAMTS5 Acan cleavage during development correlates with ascending aortic anomalies. These data indicate that the mechanism of ADAMTS5 Acan cleavage may be critical for normal aortic wall development.