长链非编码RNA FENDRR及转录因子FOXF1对婴幼儿血管瘤病理性血管生成作用效应及其机制研究

Infantile hemangioma was one of the most common benign vascular tumors in hemangioma, and neovascularization was one of the significant mechanisms. Long non-coding RNAs（lncRNA）could regulate genetic expression in transcriptional, epigenetic, translational levels. During the neovascularization, lncRNA MIAT & MALAT1 was up regulated in diabetic retinopathy with function of promoting angiogenesis. Our previous work indicated that lncRNA was dysregualted in infantile hemangioma through microarray analysis, and lncRNA FENDRR was the most upper regulated molecule, and its adjacent transcriptional factor FOXF1 was also up regulated. This project aimed to confirm the expression of FENDRR & FOXF1 in more hemangioma tissues through QPCR analysis; to explore the cellular function of FENDRR on the basis of shRNA, sgRNA & over expression vector construction and cell transfection; and to explore the direct and indirect mechanism of FENDRR through RIP and proteomics analysis. To explore the function of FENDRR in vivo, we construct the hemangioma nude mice model and hyperoxia induced retinal neovascularization mice model. This subject could uncover the important role of lncRNA in the pathogenesis in hemangioma especially in the phases of angiogenesis, otherwise, this subject could provide significant evidence for other angiogenic disease, and finally, lncRNA may serve as the therapeutic target in the angiogenesis related disease.

婴幼儿血管瘤是婴幼儿时期常见良性肿瘤，新生血管生成是其重要病理机制。长链非编码RNA（lncRNA）可在转录、表观遗传、翻译层面调控基因表达。在新生血管生成中，lncRNA MIAT及MALAT1在糖尿病视网膜病变中促进新生血管生成。我们前期工作发现血管瘤瘤体lncRNA存在显著差异表达，其中FENDRR及其邻近转录因子FOXF1表达上调明显。本课题拟采用扩大样本采用QPCR法验证FENDRR与FOXF1表达量；采用shRNA，sgRNA及过表达法构建慢病毒转染细胞后探究FENDRR及FOXF1的细胞学功能；采用蛋白质组学及RNA免疫共沉淀实验探究FENDRR调控血管生成间接和直接机制；采用血管瘤动物模型及高氧诱发小鼠视网膜新生血管生成模型探究FENDRR功能。探究FENDRR与新生血管生成相关机制有可能揭开lncRNA在血管瘤发病中的作用，为新生血管生成性疾病的靶向治疗靶点。

婴幼儿血管瘤（IH）是婴幼儿十七最常见的软组织良性病变，约占良性占位性病变的7%,位于重要部位或快速生长的IH可导致严重的外形及功能损害。目前其分类和鉴别诊断混乱，治疗选择不及时和不规范，可给患者留下终生遗憾。既往研究发现非编码RNA在IH的疾病进程中至关重要，一些miRNA还可作为IH的潜在诊断标志物及预后指标。因此，完善IH的表观遗传机制显得尤为重要。首先我们立足于之前IH中lncRNA测序结果，证实了増生期IH中，转录因子FOXF1及lncRNA FENDRR显著高表达，且FENDRR与FOXF1的表达量存在正相关。探究了FENDRR与新生血管生成相关机制，有可能揭开lncRNA在婴幼儿血管瘤发病中的作用，为新生血管生成性疾病的祀向治疗提供靶点。随后收集了在此期间内手术切除的IH组织标本，分别进行了circRNA表达谱分析、6-mA DNA 修饰水平比对以及染色质开放的特征分析。该系列研究于世界首次对IH中的circRNA、6-mA DNA 修饰和染色质开放特征进行分析，发现circRNA、6-mA DNA 修饰和染色质开放程度在IH的发生发展中均起到重要作用。该系列研究丰富了IHs研究的数据库，可供研究者们联合蛋白质修饰、甲基化等数据库进一步分析。对IH发病机制的探索是为了更好的治疗，普萘洛尔是治疗IH的一线药物。近年来，很多医师着重于普萘洛尔治 IHs停药后的复发的现象和机制、普萘洛尔对婴幼儿的近期副作用危险因素的分析、普萘洛尔对婴幼儿人群远期副作用随访等，并继续寻找优于普萘洛尔的药物。本研究对口服普萘洛尔治疗IHs的有效性、副作用、复发后的治疗等做出研究，并对IHs的治疗进行思考总结，为临床治疗婴幼儿血管瘤提供补充信息。