Joladarashi Darukeshwara, Garikipati Venkata Naga Srikanth, Thandavarayan Rajarajan A, Verma Suresh K, Mackie Alexander R, Khan Mohsin, Gumpert Anna M, Bhimaraj Arvind, Youker Keith A, Uribe Cesar, Suresh Babu Sahana, Jeyabal Prince, Kishore Raj, Krishnamurthy Prasanna
Department of Cardiovascular Sciences, Center for Cardiovascular Regeneration, Houston Methodist Research Institute, Houston, Texas.
Center for Translational Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.
J Am Coll Cardiol. 2015 Nov 17;66(20):2214-2226. doi: 10.1016/j.jacc.2015.09.009.
MicroRNA (miR) dysregulation in the myocardium has been implicated in cardiac remodeling after injury or stress.
The aim of this study was to explore the role of miR in human CD34(+) cell (hCD34(+)) dysfunction in vivo after transplantation into the myocardium under ischemia-reperfusion (I-R) conditions.
In response to inflammatory stimuli, the miR array profile of endothelial progenitor cells was analyzed using a polymerase chain reaction-based miR microarray. miR-377 expression was assessed in myocardial tissue from human patients with heart failure (HF). We investigated the effect of miR-377 inhibition on an hCD34(+) cell angiogenic proteome profile in vitro and on cardiac repair and function after I-R injury in immunodeficient mice.
The miR array data from endothelial progenitor cells in response to inflammatory stimuli indicated changes in numerous miR, with a robust decrease in the levels of miR-377. Human cardiac biopsies from patients with HF showed significant increases in miR-377 expression compared with nonfailing control hearts. The proteome profile of hCD34(+) cells transfected with miR-377 mimics showed significant decrease in the levels of proangiogenic proteins versus nonspecific control-transfected cells. We also validated that serine/threonine kinase 35 is a target of miR-377 using a dual luciferase reporter assay. In a mouse model of myocardial I-R, intramyocardial transplantation of miR-377 silenced hCD34(+) cells in immunodeficient mice, promoting neovascularization (at 28 days, post-I-R) and lower interstitial fibrosis, leading to improved left ventricular function.
These findings indicate that HF increased miR-377 expression in the myocardium, which is detrimental to stem cell function, and transplantation of miR-377 knockdown hCD34(+) cells into ischemic myocardium promoted their angiogenic ability, attenuating left ventricular remodeling and cardiac fibrosis.
心肌中的微小RNA（miR）失调与损伤或应激后的心脏重塑有关。
本研究旨在探讨miR在缺血再灌注（I-R）条件下移植到心肌中的人CD34+细胞（hCD34+）功能障碍中的作用。
针对炎症刺激，使用基于聚合酶链反应的miR微阵列分析内皮祖细胞的miR阵列谱。评估了心力衰竭（HF）患者心肌组织中miR-377的表达。我们研究了miR-377抑制对体外hCD34+细胞血管生成蛋白质组谱以及免疫缺陷小鼠I-R损伤后心脏修复和功能的影响。
内皮祖细胞对炎症刺激的miR阵列数据表明众多miR发生变化，其中miR-377水平显著降低。与非衰竭对照心脏相比，HF患者的人体心脏活检显示miR-377表达显著增加。与非特异性对照转染细胞相比，用miR-377模拟物转染的hCD34+细胞的蛋白质组谱显示促血管生成蛋白水平显著降低。我们还使用双荧光素酶报告基因测定法验证了丝氨酸/苏氨酸激酶35是miR-377的靶标。在心肌I-R小鼠模型中，在免疫缺陷小鼠心肌内移植miR-377沉默的hCD34+细胞，促进了新生血管形成（I-R后28天）并降低了间质纤维化，从而改善了左心室功能。
这些发现表明，HF增加了心肌中miR-377的表达，这对干细胞功能有害，将miR-377敲低的hCD34+细胞移植到缺血心肌中可促进其血管生成能力，减轻左心室重塑和心脏纤维化。
