Systemic injection of AAV9 carrying a periostin promoter targets gene expression to a myofibroblast-like lineage in mouse hearts after reperfused myocardial infarction
Systemic injection of AAV9 carrying a periostin promoter targets gene expression to a myofibroblast-like lineage in mouse hearts after reperfused myocardial infarction
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DOI：
10.1038/gt.2016.20
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发表时间：
2016-05-01
期刊：
影响因子：
5.1
通讯作者：
French, B. A.
中科院分区：
文献类型：
--
作者：
Piras, B. A.;Tian, Y.;French, B. A.
腺相关病毒（AAV）已被用于指导基因转移到各种组织，包括心脏、肝脏、骨骼肌、大脑、肾脏和肺，但它以前没有被证明能有效地靶向体内的成纤维细胞，包括心脏成纤维细胞。我们使用改良的骨膜蛋白启动子构建了表达盒，以驱动心肌成纤维细胞样谱系中的基因表达，仅偶尔溢出到心肌细胞样细胞中。我们比较了AAV血清型6和血清型9，发现在心肌梗死（MI）后全身注射载体时，基因表达强劲，而在健康的非梗死小鼠中表达很少。与AAV6相比，AAV9在更多的细胞中表达，通过荧光素酶和cre激活的绿色荧光蛋白表达评估，报告基因在心肌梗死后5至62天在心肌梗死区和边界区可见。尽管常见的肌成纤维细胞标记物的表达丰度较低，但大多数靶向细胞表达肌球蛋白IIb，这是一种胚胎形式的平滑肌肌球蛋白重链，以前与心肌再灌注后的肌成纤维细胞有关。这项研究首次证明了心肌梗死后小鼠心脏中aav介导的潜在的新型肌成纤维细胞样细胞系的表达，可能为心肌梗死存活患者的基因治疗开辟新的途径。
Adeno-associated virus (AAV) has been used to direct gene transfer to a variety of tissues, including heart, liver, skeletal muscle, brain, kidney and lung, but it has not previously been shown to effectively target fibroblasts in vivo, including cardiac fibroblasts. We constructed expression cassettes using a modified periostin promoter to drive gene expression in a cardiac myofibroblast-like lineage, with only occasional spillover into cardiomyocyte-like cells. We compared AAV serotypes 6 and 9 and found robust gene expression when the vectors were delivered by systemic injection after myocardial infarction (MI), with little expression in healthy, non-infarcted mice. AAV9 provided expression in a greater number of cells than AAV6, with reporter gene expression visible in the cardiac infarct and border zones from 5 to 62 days post MI, as assessed by luciferase and Cre-activated green fluorescent protein expression. Although common myofibroblast markers were expressed in low abundance, most of the targeted cells expressed myosin IIb, an embryonic form of smooth muscle myosin heavy chain that has previously been associated with myofibroblasts after reperfused MI. This study is the first to demonstrate AAV-mediated expression in a potentially novel myofibroblast-like lineage in mouse hearts post MI and may open new avenues of gene therapy to treat patients surviving MI.