N-Cadherin Overexpression Mobilizes the Protective Effects of Mesenchymal Stromal Cells Against Ischemic Heart Injury Through a β-Catenin-Dependent Manner
N-Cadherin Overexpression Mobilizes the Protective Effects of Mesenchymal Stromal Cells Against Ischemic Heart Injury Through a β-Catenin-Dependent Manner
复制标题
N-钙粘蛋白过表达通过β-连环蛋白依赖性方式调动间充质基质细胞对缺血性心脏损伤的保护作用
DOI：
10.1161/circresaha.119.315806
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发表时间：
2020-03-27
影响因子：
20.1
通讯作者：
Tao, Ling
中科院分区：
文献类型：
--
作者：
Yan, Wenjun;Lin, Chen;Tao, Ling
理论基础：以间充质基质细胞为基础的治疗有望对抗缺血性心力衰竭。然而，由于细胞存留率低，旁分泌功能差，其疗效受到限制。N-钙粘蛋白是一种能够增强细胞-细胞黏附的跨膜蛋白，最近才引起干细胞生物学领域的关注。目的：研究N-钙粘蛋白是否以及如何调节间充质干细胞的滞留和对缺血性心力衰竭的心脏保护能力。方法和结果：将携带N-钙粘素、T-钙粘素的腺病毒或对照组腺病毒导入成年小鼠脂肪组织来源的间充质干细胞(ADSC)。在心肌梗死(MI)或心肌缺血/再灌注后即刻将CM-DiI标记的ADSC注射到梗死区的3个部位。检测ADSC存留/存活、心肌细胞凋亡/增殖、毛细血管密度、心肌纤维化和心功能。用发现驱动/因果分析来确定分子机制。与转染腺病毒对照的ADSC相比，高表达N-钙粘蛋白(而不是T-钙粘蛋白)可显著增加移植ADSC的存活/滞留至心肌梗死后7天。组织学分析显示，心肌梗死后3d，转染腺病毒N-钙粘素的ADSC可明显保持心肌毛细血管密度，促进心肌细胞增殖，适度减少心肌细胞凋亡。更重要的是，转染腺病毒-N-钙粘素的ADSC(而不是转染腺病毒-T-钙粘素的ADSC)显著增加了心肌梗死和心肌缺血/再灌注小鼠的左心室射血分数，减少了心肌纤维化。体外实验表明，N-钙粘附素过表达可促进ADSC与心肌细胞的黏附和迁移，增强其促进血管生成和心肌细胞增殖的能力。基质金属肽酶-10/13和肝细胞生长因子上调与N-钙粘蛋白对ADSC迁移和旁分泌血管生成的影响有关。N-钙粘附素过表达通过释放HGF促进心肌细胞增殖。在机制上，N-钙粘蛋白的过度表达显著增加N-钙粘素/β-连环蛋白复合体的形成和核内活跃的β-连环蛋白水平。结论：本研究首次证实了N-钙粘素过表达可增强间充质细胞对缺血性心力衰竭的保护作用，其机制可能是通过β-连环蛋白介导的基质细胞表达和分泌，促进ADSC/心肌细胞的黏附和ADSC的滞留。
Rationale:Mesenchymal stromal cell-based therapy is promising against ischemic heart failure. However, its efficacy is limited due to low cell retention and poor paracrine function. A transmembrane protein capable of enhancing cell-cell adhesion, N-cadherin garnered attention in the field of stem cell biology only recently.Objective:The current study investigates whether and how N-cadherin may regulate mesenchymal stromal cells retention and cardioprotective capability against ischemic heart failure.Methods and Results:Adult mice-derived adipose tissue-derived mesenchymal stromal cells (ADSC) were transfected with adenovirus harboring N-cadherin, T-cadherin, or control adenovirus. CM-DiI-labeled ADSC were intramyocardially injected into the infarct border zone at 3 sites immediately after myocardial infarction (MI) or myocardial ischemia/reperfusion. ADSC retention/survival, cardiomyocyte apoptosis/proliferation, capillary density, cardiac fibrosis, and cardiac function were determined. Discovery-driven/cause-effect analysis was used to determine the molecular mechanisms. Compared with ADSC transfected with adenovirus-control, N-cadherin overexpression (but not T-cadherin) markedly increased engrafted ADSC survival/retention up to 7 days post-MI. Histological analysis revealed that ADSC transfected with adenovirus-N-cadherin significantly preserved capillary density and increased cardiomyocyte proliferation and moderately reduced cardiomyocyte apoptosis 3 days post-MI. More importantly, ADSC transfected with adenovirus-N-cadherin (but not ADSC transfected with adenovirus-T-cadherin) significantly increased left ventricular ejection fraction and reduced fibrosis in both MI and myocardial ischemia/reperfusion mice. In vitro experiments demonstrated that N-cadherin overexpression promoted ADSC-cardiomyocyte adhesion and ADSC migration, enhancing their capability to increase angiogenesis and cardiomyocyte proliferation. MMP (matrix metallopeptidases)-10/13 and HGF (hepatocyte growth factor) upregulation is responsible for N-cadherin's effect upon ADSC migration and paracrine angiogenesis. N-cadherin overexpression promotes cardiomyocyte proliferation by HGF release. Mechanistically, N-cadherin overexpression significantly increased N-cadherin/beta-catenin complex formation and active beta-catenin levels in the nucleus. beta-catenin knockdown abolished N-cadherin overexpression-induced MMP-10, MMP-13, and HGF expression and blocked the cellular actions and cardioprotective effects of ADSC overexpressing N-cadherin.Conclusions:We demonstrate for the first time that N-cadherin overexpression enhances mesenchymal stromal cells-protective effects against ischemic heart failure via beta-catenin-mediated MMP-10/MMP-13/HGF expression and production, promoting ADSC/cardiomyocyte adhesion and ADSC retention.