microRNA-29a Regulates ADAM12 Through Direct Interaction With ADAM12 mRNA and Modulates Postischemic Perfusion Recovery.
microRNA-29a Regulates ADAM12 Through Direct Interaction With ADAM12 mRNA and Modulates Postischemic Perfusion Recovery.
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DOI：
10.1161/jaha.122.025727
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发表时间：
2022-08-16
影响因子：
5.4
通讯作者：
Dokun, Ayotunde O.
中科院分区：
文献类型：
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作者：
Lamin, Victor;Verry, Joseph;Dokun, Olumayowa S.;Kronemberger, Ana;Wong, Thomas;Lira, Vitor A.;Dokun, Ayotunde O.
外周动脉疾病是由心脏外血管的动脉粥样硬化闭塞引起的，最常见的是影响下肢血管。血管生成是缺血后适应的一部分，参与恢复外周动脉疾病的血流。以前，在外周动脉疾病的小鼠后肢缺血模型中，我们确定了ADAM 12（去整合素和金属蛋白酶基因12）作为缺血后灌注恢复的关键遗传修饰因子。然而，对缺血中的ADAM 12调节知之甚少。MicroRNA是一类小的、非编码的单链RNA，主要通过信使RNA（mRNA）的转录抑制来调节基因表达。我们发现microRNA-29 a（miR-29 a）在糖尿病和缺血的情况下调节ADAM 12的表达。然而，miR-29 a如何调节ADAM 12尚不清楚。此外，miR-29 a调节在非糖尿病环境中的生理作用尚不清楚。我们在缺血小鼠腓肠肌和胫骨前肌中过表达或抑制miR-29 a，并量化了对灌注恢复、ADAM 12表达、血管生成和骨骼肌再生的影响。此外，使用基于RNA免疫沉淀的抗miR竞争性测定，我们研究了小鼠微血管内皮细胞、骨骼肌和人内皮细胞裂解物中miR-29 a和ADAM 12 mRNA的相互作用。缺血小鼠后肢中miR-29 a的异位表达降低了ADAM 12 mRNA的表达，增加了骨骼肌损伤，降低了骨骼肌功能，降低了血管生成和灌注恢复，对后肢缺血后骨骼肌再生和肌纤维横截面积没有影响。基于RNA免疫沉淀的抗miR竞争性测定研究显示，miR-29 a Ekomir以剂量依赖性方式从AGO-2（Argonaut-2）复合物中置换miR-29 a和ADAM 12 mRNA。综上所述，数据显示miR-29 a通过直接结合其mRNA来抑制ADAM 12的表达，导致骨骼肌功能受损、血管生成和灌注不良。因此，如糖尿病和衰老中所见，miR-29 a水平升高可能导致血管病理学，并且miR-29 a的调节可能是治疗靶点。
Peripheral artery disease is caused by atherosclerotic occlusion of vessels outside the heart and most commonly affects vessels of the lower extremities. Angiogenesis is a part of the postischemic adaptation involved in restoring blood flow in peripheral artery disease. Previously, in a murine hind limb ischemia model of peripheral artery disease, we identified ADAM12 (a disintegrin and metalloproteinase gene 12) as a key genetic modifier of postischemic perfusion recovery. However, less is known about ADAM12 regulation in ischemia. MicroRNAs are a class of small, noncoding, single‐stranded RNAs that regulate gene expression primarily through transcriptional repression of messenger RNA (mRNA). We showed microRNA‐29a (miR‐29a) modulates ADAM12 expression in the setting of diabetes and ischemia. However, how miR‐29a modulates ADAM12 is not known. Moreover, the physiological effects of miR‐29a modulation in a nondiabetic setting is not known. We overexpressed or inhibited miR‐29a in ischemic mouse gastrocnemius and tibialis anterior muscles, and quantified the effect on perfusion recovery, ADAM12 expression, angiogenesis, and skeletal muscle regeneration. In addition, using RNA immunoprecipitation–based anti‐miR competitive assay, we investigated the interaction of miR‐29a and ADAM12 mRNA in mouse microvascular endothelial cell, skeletal muscle, and human endothelial cell lysates. Ectopic expression of miR‐29a in ischemic mouse hind limbs decreased ADAM12 mRNA expression, increased skeletal muscle injury, decreased skeletal muscle function, and decreased angiogenesis and perfusion recovery, with no effect on skeletal muscle regeneration and myofiber cross‐sectional area following hind limb ischemia. RNA immunoprecipitation–based anti‐miR competitive assay studies showed miR‐29a antagomir displaced miR‐29a and ADAM12 mRNA from the AGO‐2 (Argonaut‐2) complex in a dose dependent manner. Taken together, the data show miR‐29a suppresses ADAM12 expression by directly binding to its mRNA, resulting in impaired skeletal muscle function, angiogenesis, and poor perfusion. Hence, elevated levels of miR‐29a, as seen in diabetes and aging, likely contribute to vascular pathology, and modulation of miR‐29a could be a therapeutic target.