Novel role of the lncRNA NEAT1 in smooth muscle phenotypic modulation

lncRNA NEAT1在平滑肌表型调节中的新作用

• 批准号: 9251903
• 负责人: Jiliang Zhou
• 金额: $ 38万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2020-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9251903
• 关键词: American; Angioplasty; Apolipoprotein E; Architecture; Area; Arterial Injury; Arteries; Atherosclerosis; Attenuated; Binding; Biological Assay; Biotin; Blood Vessels; CREB1 gene; Cardiovascular Diseases; Cardiovascular system; Carotid Arteries; Cell Proliferation; Chromatin; Complex; Contractile Proteins; Cyclic AMP Response Element; Data; Development; Disease; Down-Regulation; Evolution; Family; Fatty acid glycerol esters; Fishes; Foundations; Gel; Gene Expression; Genes; Genetic Transcription; Genomic DNA; Goals; Growth Factor; Histone H3; Histones; Human; Hyperplasia; Hypertension; IL8 gene; Immunohistochemistry; In Vitro; Injury; Knockout Mice; Lesion; Luciferases; Lysine; Malignant neoplasm of prostate; Mammals; Maps; Mediating; Methylation; Methyltransferase; Mitogens; Muscle Cells; Nuclear; Pathologic; Pathology; Pharmacology; Phenotype; Play; Prevention; Procedures; Production; Proteins; Quantitative Reverse Transcriptase PCR; RNA; RNA Binding; Rattus; Regulator Genes; Reporter; Research; Role; Signal Pathway; Smooth Muscle; Stimulus; Testing; Therapeutic; Therapeutic Agents; Tissues; Transactivation; Transcript; Untranslated RNA; Vascular Diseases; Vascular Smooth Muscle; Virus Diseases; WD Repeat; Western Blotting; Work; cell motility; design; experimental study; femoral artery; gain of function; in vivo; inhibitor/antagonist; injured; insight; loss of function; malignant breast neoplasm; migration; mortality; mutant; novel; platelet-derived growth factor BB; prevent; promoter; public health relevance; response; restenosis; transcription factor; tumorigenesis

 DESCRIPTION (provided by applicant): The overall goal of the proposed research is to determine the previously unrecognized role of the (long non-coding RNA) lncRNA NEAT1 (nuclear paraspeckle assembly transcript 1) and its novel underlying mechanism in regulating the phenotypic switching of vascular smooth muscle cells (VSMCs). Many vascular diseases in humans, such as intimal hyperplasia post-angioplasty, are largely dependent upon VSMC phenotypic switching from a contractile to a "synthetic" phenotype that is associated with reduced smooth muscle-specific gene expression and increased cell proliferation and migration. Therefore, unraveling the mechanisms for the VSMC phenotypic switching is crucial for understanding the pathology of VSMC related vascular diseases and ultimately designing therapeutic agents for treatment. Emerging evidences demonstrate that lncRNAs represent a novel class of regulators for gene expression. In an effort to search for lncRNAs involved in the vascular injury, we conducted a large-scale lncRNA array screen using the rat carotid artery injured by a balloon denudation, a procedure resembling angioplasty in humans. This screen revealed that the lncRNA NEAT1 is induced in response to the arterial injury in vivo. Furthermore, expression of NEAT1 is also induced upon the stimulation of smooth muscle phenotypic modulation in vitro. Loss- and gain-of-function NEAT1 revealed that NEAT1 not only increases proliferation and migration of VSMCs but also decreases expression of smooth muscle-specific contractile proteins. Therefore, experiments described in this proposal will test the hypothesis that, the induction of NEAT1 plays a critical role in promoting smooth muscle phenotypic switching. In Aim 1, we will determine the role of NEAT1 in neointima hyperplasia following arterial injury. We will perform femoral artery wire injuries in the control and NEAT1 knock out mice and assess the effects of NEAT1 KO on the progression of neointima hyperplasia. In Aim 2, we will determine the mechanism by which NEAT1 abrogates smooth muscle-specific gene expression. Specifically we will evaluate the role of WDR5, a critical histone modifier for inducing an active form of chromatin, in mediating NEAT1 function in VSMCs. In Aim 3, we will determine the mechanism by which the transcription of NEAT1 is up-regulated upon the stimulation of smooth muscle phenotypic modulation by testing the role of an evolutionally conserved cAMP response element identified in the NEAT1 proximal promoter. Completion of these studies will provide novel insights into the mechanisms controlling smooth muscle phenotypic switching and determine if preventing the induction of NEAT1 may be an attractive therapeutic strategy for ameliorating occlusive vascular diseases.

 描述（由适用提供）：拟议的研究的总体目标是确定（长的非编码RNA）lncRNA neat1（核副羊皮纸组装转录本）及其在确定血管平滑肌细胞的表型切换方面的新基础机制（VSMC）。人类中的许多血管疾病，例如血管成形术后内膜增生，在很大程度上取决于VSMC表型从收缩的转变为“合成”表型的转换，这些转变与平滑肌特异性基因表达减少和增加细胞增殖和迁移有关。因此，阐明VSMC表型转换的机制对于理解VSMC相关血管疾病的病理和最终设计治疗剂以进行治疗至关重要。新兴的证据表明，lncRNA代表了一种新型的基因表达调节剂。为了寻找涉及血管损伤的LNCRNA，我们使用球囊剥落受伤的大鼠颈动脉进行了大规模的LncRNA阵列筛网，这是一种类似于人类血管成形术的手术。该屏幕表明，lncRNA neat1响应于体内动脉损伤。此外，在刺激平滑肌表型调节中，Neat1的表达也被诱导，在功能障碍和功能获得neat1中表明，Neat1不仅增加了VSMC的增殖和迁移，而且还会降低平滑肌特异性收缩蛋白的表达。因此，本提案中描述的实验将检验以下假设：NEAT1的诱导在促进平滑肌表型转换中起关键作用。在AIM 1中，我们将确定Neat1在动脉损伤后Neintima增生中的作用。我们将在对照中进行股动脉损伤，而Neat1敲除小鼠，并评估Neat1 KO对新内膜增生的进展的影响。在AIM 2中，我们将确定Neat1废除平滑肌特异性基因表达的机制。具体而言，我们将评估WDR5的作用，WDR5是诱导染色质的活性形式的关键组蛋白修饰剂，在介导VSMC中的Neat1功能中。 AIM 3，我们将通过测试Neat1代理启动子中确定的Evolvely组成的CAMP响应元件的作用来确定neat1的转录在刺激平滑肌表型调制时上调的机制。这些研究的完成将为控制平滑肌表型转换的机制提供新的见解，并确定防止Neat1的诱导是否可能是改善闭塞性血管疾病的有吸引力的治疗策略。