AIMS Cardiac fibrosis is a major cause of heart failure, and mediated by the differentiation of cardiac fibroblasts into myofibroblasts. However, limited tools are available to block cardiac fibrosis. ADAMTS16 is a member of the ADAMTS superfamily of extracellular protease enzymes involved in extracellular matrix degradation and remodeling. In this study, we aimed to establish ADAMTS16 as a key regulator of cardiac fibrosis. METHODS AND RESULTS Western blot and qRT-PCR analyses demonstrated that ADAMTS16 was significantly up-regulated in mice with transverse aortic constriction (TAC) associated with left ventricular hypertrophy and heart failure, which was correlated with increased expression of Mmp2, Mmp9, Col1a1 and Col3a1. Overexpression of ADAMTS16 accelerated the AngII-induced activation of cardiac fibroblasts into myofibroblasts. Protein structural analysis and co-immunoprecipitation revealed that ADAMTS16 interacted with the latency-associated peptide (LAP)-TGF-β via a RRFR motif. Overexpression of ADAMTS16 induced the activation of TGF-β in cardiac fibroblasts, however, the effects were blocked by a mutation of the RRFR motif to IIFI, knockdown of Adamts16 expression or a TGF-β-neutralizing antibody (ΝAb). The RRFR tetrapeptide, but not control IIFI peptide, blocked the interaction between ADAMTS16 and LAP-TGF-β, and accelerated the activation of TGF-β in cardiac fibroblasts. In TAC mice, the RRFR tetrapeptide aggravated cardiac fibrosis and hypertrophy by upregulation of extracellular matrix proteins, activation of TGF-β, and increased SMAD2/SMAD3 signaling, however, the effects were blocked by TGF-β-NAb. CONCLUSIONS ADAMTS16 promotes cardiac fibrosis, cardiac hypertrophy and heart failure by facilitating cardiac fibroblasts activation via interacting with and activating LAP-TGF-β signaling. The RRFR motif of ADAMTS16 disrupts the interaction between ADAMTS16 and LAP-TGF-β, activates TGF-β, and aggravated cardiac fibrosis and hypertrophy. This study identifies a novel regulator of TGF-β signaling and cardiac fibrosis, and provides a new target for the development of therapeutic treatment of cardiac fibrosis and heart failure.

中文翻译：

---

ADAMTS16激活潜在的TGF-beta，加速压力超负荷心脏的纤维化和功能障碍。

AIMS心脏纤维化是心力衰竭的主要原因，由心脏成纤维细胞分化为成肌纤维细胞介导。但是，可用有限的工具来阻止心脏纤维化。ADAMTS16是参与细胞外基质降解和重塑的细胞外蛋白酶ADAMTS超家族的成员。在这项研究中，我们旨在将ADAMTS16建立为心脏纤维化的关键调节剂。方法和结果Western blot和qRT-PCR分析表明，ADAMTS16在伴有左心室肥大和心力衰竭的横向主动脉缩窄（TAC）的小鼠中显着上调，这与Mmp2，Mmp9，Col1a1和Col3a1的表达增加有关。ADAMTS16的过表达加速了AngII诱导的心脏成纤维细胞活化为成肌纤维细胞。蛋白质结构分析和免疫共沉淀显示ADAMTS16通过RRFR基序与潜伏期相关肽（LAP）-TGF-β相互作用。ADAMTS16的过表达诱导了心脏成纤维细胞中TGF-β的激活，但是，这种作用被RRFR基序突变为IIFI，Adamts16表达的敲低或TGF-β中和抗体（ΝAb）所阻断。RRFR四肽而非对照IIFI肽阻断了ADAMTS16与LAP-TGF-β之间的相互作用，并加速了心脏成纤维细胞中TGF-β的活化。在TAC小鼠中，RRFR四肽通过细胞外基质蛋白的上调，TGF-β的活化和SMAD2 / SMAD3信号转导的增加而加重了心脏纤维化和肥大，但是，TGF-β-NAb阻断了这种作用。结论ADAMTS16促进心脏纤维化，通过与LAP-TGF-β信号相互作用并激活LAP-TGF-β信号来促进心脏成纤维细胞的活化，从而导致心脏肥大和心力衰竭。ADAMTS16的RRFR基序破坏了ADAMTS16与LAP-TGF-β之间的相互作用，激活了TGF-β，并加剧了心脏纤维化和肥大。这项研究确定了一种新型的TGF-β信号传导和心脏纤维化调节剂，并为开发治疗心脏纤维化和心力衰竭的新靶标提供了新的靶标。