lncRNA-OVM1调控α-SMA在眼眶静脉畸形形成中的机制研究

Orbital venous malformations is (OVM) is a common eye disease which is severe and intractable. Its main pathological features is having decreased elasticity of lesion blood vessel and expansive vascular wall. Preliminary study found: 1. The alpha smooth muscle actin (α-SMA) expression level which is the key blood vessels elastic components in OVM is significantly reduced. 2. Various of long non-coding RNAs (lncRNAs) are over expressed in OVM and lncRNA - OVM1 is the most obvious one. 3. α-SMA expression level can be improved by inhibiting lncRNA - OVM1 in OVM. Above all, we suggest that abnormally up-regulated lncRNA - OVM1 inhibited the expression level of α-SMA and leading the decreasion of blood vessel elasticity and expansion of vascular wall. This study will use the primary cultured OVM smooth muscle cells and human umbilical vein smooth muscle cell to study the mechanism of lncRNA - OVM1 regulating α-SMA by the technology of oligonucleotide precipitation (CHOP), RNA oligonucleotides chromosome conformation capture (R3C) and so on. We will explore the key factor of α-SMA down regulated by lncRNA - OVM1 and validate the function of the key factor, so as to provide theoretical basis for the pathogenesis of OVM and provide a potential target for the OVM clinical study.

眼眶静脉畸形（OVM）是常见眼眶病，危害大、治疗棘手。OVM主要病理特征为病变血管弹性降低、管壁扩张变薄。前期研究发现：1、OVM中关键血管弹力成分α-平滑肌肌动蛋白（α-SMA）表达显著减少；2、OVM中多条长非编码RNA（lncRNA）表达升高，其中lncRNA-OVM1最明显；3、OVM中lncRNA-OVM1表达位于OVM平滑肌细胞核内。据此推测：异常升高的lncRNA-OVM1抑制α-SMA表达，使血管平滑肌弹力下降，血管扩张变薄，最终促进OVM形成。本研究将借助原代培养的OVM平滑肌细胞和人脐静脉平滑肌细胞系，通过观察抑制或过表达lncRNA-OVM1对α-SMA的影响来验证该假设，并利用寡核苷酸沉淀（CHOP）、RNA染色体构象捕获（R3C）等技术，探究lncRNA-OVM1调控α-SMA的作用机制，以期部分揭示OVM形成的分子生物学机制，为OVM临床治疗研究提供潜在靶点。

眼眶静脉畸形（OVM）是一种以包膜清晰、血管通道众多为特征的良性血管疾病，是常见的原发性眼眶病变。然而，OVMs的发病机制和诊断标志物尚未见报道。为了确定眼眶静脉畸形的发病机制，我们在眼眶静脉畸形样本和正常血管组织中进行了cDNA微阵列分析。实时聚合酶链反应进一步证实了OVM队列中改变的关键信号通路的表达。为探讨OVM的诊断标志物，采用血管抗体芯片技术进行蛋白质组学筛查。酶联免疫吸附试验（ELISA）进一步证实了OVM队列中这些因子的表达。转录组测序显示自噬，VEGF信号通路升高。在OVM临床样本中，Hippo、Wnt、Hedgehod和血管平滑肌收缩信号通路均显著下调。OVM患者血浆EGF（p<0.001）和瘦素（p<0.01）水平明显升高。本文首次揭示了OVM的转录背景和诊断性血浆标志物，为OVM的发病机制提供了新的认识，有助于OVM的早期诊断。上述主要内容已被国际眼科研究协会（International Society for Eye Research）会刊Exp Eye Res接收发表，另有一篇相关论文Bone morphogenetic protein 9 delays occurrence of venous malformation by strengthening vessel walls and maintaining endothelial quiescence已经投稿。同时在具体实验实施过程中，发现OVM病人相对较少，细胞株和动物模型建立比较困难，而眼部血管增生异常的类似症状病人较多，有商品化的细胞株和成熟的动物模型等。利用同样的研究思路，我们选择转录-蛋白组联合测序手段，也开展了靶向基因测序筛选、RNA甲基化、长非编码RNA等研究，并且发表了International Journal of Cancer、Mol Cancer等7篇基金标注SCI论文。研究期间培养博士两名，硕士两名，举办国际学术大会1次，国内学术大会两次，受邀至世界眼科学大会做专题汇报一次。在项目完成期间申请人作为第2 完成人获得2018年国家科技进步奖二等奖。