MicroRNA-133a engineered mesenchymal stem cells augment cardiac function and cell survival in the infarct heart.
MicroRNA-133a engineered mesenchymal stem cells augment cardiac function and cell survival in the infarct heart.
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DOI：
10.1097/fjc.0000000000000183
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发表时间：
2015-03
影响因子：
3
通讯作者：
Khan M
中科院分区：
文献类型：
--
作者：
Dakhlallah D;Zhang J;Yu L;Marsh CB;Angelos MG;Khan M
心血管疾病是美国发病率和死亡率的头号原因。心血管疾病最常见的表现是心肌梗死（MI），最终可导致充血性心力衰竭（CHF）。细胞疗法（心肌成形术）是一种新的潜在的治疗方法，为受损的心脏。最近的临床前和临床研究表明，间充质干细胞（MSC）是一种有前途的细胞类型，用于心肌成形术的应用。然而，一个主要的限制是移植的干细胞在梗死心脏中的存活率很低。miR-133 a是心肌中大量表达的microRNA，在MI患者中下调。我们假设使用microRNA模拟物（双链寡核苷酸）重编程MSC将改善受损心脏中干细胞的存活率。用miR-133 a模拟物和miR-133 a抑制剂转染MSC，并通过qRT-PCR测量miR-133 a的水平。使大鼠心脏经受MI，并将用miR-133 a模拟物或miR-133 a模拟物转染的MSC植入缺血心脏中。MI后4周，检测心脏功能、心肌纤维化、miR-133 a水平和凋亡相关基因Apaf-1、Capase-9和Caspase-3。我们发现用miR-133 a模拟物转染MSC改善了MSC的存活，如通过MTT测定所确定的。类似地，当与仅MSC或MI组相比时，miR-133 a模拟物转染的MSC在经受MI的大鼠心脏中的移植导致细胞植入、心脏功能的显著增加和纤维化的减少。在分子水平上，qRT-PCR数据表明miR-133 a模拟物移植组中促凋亡基因Apaf-1、caspase-9和caspase-3的表达显著降低。此外，荧光素酶报告基因测定证实miR-133 a是Apaf-1的直接靶标。总的来说，通过miRNAs操作的干细胞生物工程可能会改善接受干细胞移植治疗心肌梗死的患者的治疗结果。
Cardiovascular disease is the number one cause of morbidity and mortality in the United States. The most common manifestation of cardiovascular disease is myocardial infarction (MI), which can ultimately lead to congestive heart failure (CHF). Cell therap (cardiomyoplasty) is a new potential therapeutic treatment alternative for the damaged heart. Recent preclinical and clinical studies have shown that mesenchymal stem cells (MSCs) are a promising cell type for cardiomyoplasty applications. However, a major limitation is the poor survival rate of transplanted stem cells in the infarcted heart. miR-133a is an abundantly expressed microRNA in the cardiac muscle and is down-regulated in patients with MI. We hypothesized that reprogramming MSCs using microRNA-mimics (double-stranded oligonucleotides) will improve survival of stem cells in the damaged heart. MSCs were transfected with miR-133a mimic and antagomirs and the levels of miR-133a were measured by qRT-PCR. Rat hearts were subjected to MI and MSCs transfected with miR-133a mimic or antagomir were implanted in the ischemic heart. Four weeks after MI, cardiac function, cardiac fibrosis, miR-133a levels and apoptosis related genes (Apaf-1, Capase-9 and Caspase-3) were measured in the heart. We found that transfecting MSCs with miR-133a mimic improves survival of MSCs as determined by the MTT assay. Similarly, transplantation of miR-133a mimic transfected MSCs in rat hearts subjected to MI led to a significant increase in cell engraftment, cardiac function and decreased fibrosis when compared with MSCs only or MI groups. At the molecular level, qRT-PCR data demonstrated a significant decrease in expression of the pro-apoptotic genes; Apaf-1, caspase-9 and caspase-3 in the miR-133a mimic transplanted group. Further, luciferase reporter assay confirmed that miR- 133a is a direct target for Apaf-1. Overall, bioengineering of stem cells through miRNAs manipulation could potentially improve the therapeutic outcome of patients undergoing stem cell transplantation for myocardial infarction.