BMP2 Gene Regulation in Calcific Aortic Valve Disease

BMP2 基因在钙化主动脉瓣疾病中的调控

基本信息

批准号：
8353323
负责人：
DOUGLAS P MORTLOCK
金额：
$ 38.44万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-23 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8353323
关键词：
3&apos Untranslated Regions Adult Animal Model Area BMP2 gene Bacterial Artificial Chromosomes Bicuspid Binding Biological Markers Biology Calcified Cells Cellular biology Cholesterol Chromosome Mapping Cis Tests Collection Consensus Conserved Sequence DNA Sequence Defect Development Diagnosis Diagnostic Diet Disease Disease Progression Disease model Early Diagnosis Electrophoretic Mobility Shift Assay Elements Enhancers FGF2 gene Fishes Future Gene Expression Regulation Genes Genetic Genetic Polymorphism Genetic Transcription Goals Individual Knockout Mice Knowledge Life Mammals Maps Medical MicroRNAs Modeling Molecular Morphogenesis Mus Mutate Operative Surgical Procedures Osteoblasts Osteogenesis Pattern Predisposition Proteins RNA RNA Sequences Reagent Regulation Regulatory Element Reporter Reporter Genes Research Research Infrastructure Shapes Signal Pathway Signal Transduction Site Staging Surveys Testing Therapeutic Therapeutic Intervention Transcript Transfection abstracting aortic valve aortic valve disorder bone bone morphogenetic protein 2 calcification chromatin immunoprecipitation feeding gene repression genetic element improved interest interstitial cell malformation mouse model new therapeutic target notch protein novel strategies osteoblast differentiation osteogenic prevent recombinase response therapeutic target transcription factor

项目摘要

DESCRIPTION (provided by applicant): Congenital valve defects promote Calcific Aortic Valve Disease (CAVD). Bone morphogenetic protein 2 (BMP2) is a key signal in valve development. In diseased adult valves, the pro-calcific BMP2 and downstream responses stimulate calcification. Thus, aberrant BMP2 patterns and levels influence CAVD indirectly by disrupting valve shape and directly by inducing ectopic bone formation. Our overall hypothesis is that controlling BMP2 levels would be an effective medical therapy for CAVD. Our strategy is to elucidate how BMP2 levels are controlled in the aortic valve during normal valve development and under conditions that promote CAVD. Aim 1. To test DNA sequences that control BMP2 synthesis at the transcriptional level in the valves of mice that model CAVD and during valve development. This aim will use an existing collection of reagents and mice that were created to survey bone regulatory sequences. One region includes a conserved bone enhancer that binds the master bone differentiation transcription factor RUNX2 and other potential regulators. We will test ECR1 and map other cis-elements that control Bmp2 expression in mice that develop CAVD when fed a high cholesterol diet and during the early stages of normal aortic valve development. Aim 2. To test RNA sequences that control BMP2 synthesis at the post-transcriptional level in developing valves and in the valves of mice that model CAVD. Both positive and negative mechanisms influence BMP2 synthesis in the aortic valve. A sequence conserved between mammals and fishes strongly represses BMP2 reporter gene repression in the developing and adult aortic valve. We will test the hypothesis that this element and its flanking RNA prevent BMP2 synthesis and ectopic bone in aortic valves in the Notch-compromised CAVD model. We will also test this regulatory mechanism during normal aortic valve development. Aim 3. To identify trans-regulatory molecules that control Bmp2 transcription through ECR1. The trans-regulatory factors that bind ECR1 may promote BMP2 synthesis in calcifying aortic valves. Molecular studies will be used to define factors interacting with the ECR1 sequence. The association of RUNX2 and Hes1 will be examined in MC3T3-E1 pre-osteoblasts that express BMP2 in response to FGF2. These findings will be confirmed in primary aortic valve interstitial cells and diseased valves from the CAVD mouse model. Aim 4. To identify trans-regulatory molecules that control BMP2 synthesis at the post- transcriptional level. We will test the influence of microRNAs that target the BMP2 transcript MC3T3-E1 cells and primary valve interstitial cells. This study will provide fundamental knowledge regarding the spatial and temporal control of Bmp2 in a new model of CAVD and in the developing aortic valve. These studies will direct future tests of anti-BMP2 therapies in CAVD. This proposal explores these Specific Areas of Research Interest: Genetics of CAVD and bicuspid aortic valve, Infrastructure (improved animal models), Cell biology (signaling pathways, mechanisms, and regulation of calcification), and Diagnosis. (End of Abstract)

描述（由申请人提供）：先天性瓣膜缺陷促进钙化主动脉瓣疾病（CAVD）。骨形态发生蛋白2（BMP2）是瓣膜发育中的关键信号。在患病的成年瓣膜中，促钙化BMP2和下游反应刺激了钙化。因此，异常的BMP2模式和水平通过破坏瓣膜形状并直接通过诱导异位骨形成而间接影响CAVD。我们的总体假设是，控制BMP2水平将是CAVD的有效药物疗法。我们的策略是阐明在正常瓣膜发育过程中以及在促进CAVD的条件下如何控制BMP2水平。目的1。测试控制BMP2合成在转录水平上的DNA序列中的小鼠瓣膜和瓣膜发育过程中。该目标将使用现有的试剂和小鼠集合来调查骨调节序列。一个区域包括一个保守的骨增强子，该骨骼增强剂结合了主骨分化转录因子Runx2和其他潜在调节剂。我们将测试ECR1并绘制其他CIS元素，这些元素控制在喂养高胆固醇饮食并在正常主动脉瓣发育的早期阶段喂养CAVD的小鼠中的BMP2表达。 AIM 2。测试在发育阀和cavd模型的小鼠阀中，在转录后水平上控制BMP2合成的RNA序列。正和负机制都会影响主动脉瓣中BMP2的合成。哺乳动物和鱼之间保守的序列强烈抑制发育中和成人主动脉瓣中BMP2报告基因抑制。我们将测试以下假设：该元素及其侧翼RNA可预防Notch启动性CAVD模型中主动脉瓣中BMP2合成和异位骨。我们还将在正常主动脉瓣发育期间测试这种调节机制。目标3。确定通过ECR1控制BMP2转录的反式调节分子。结合ECR1的反式调节因子可能会促进钙化主动脉瓣中的BMP2合成。分子研究将用于定义与ECR1序列相互作用的因素。 RUNX2和HES1的关联将在响应FGF2的MC3T3-E1前成核细胞中进行检查。这些发现将在初级主动脉瓣间隙细胞和CAVD小鼠模型中患病的瓣膜中得到证实。目标4。确定在转录后水平控制BMP2合成的反式调节分子。我们将测试针对BMP2转录本MC3T3-E1细胞和原代瓣膜细胞的microRNA的影响。这项研究将提供有关BMP2在新的CAVD和发育中的主动脉瓣中的空间和时间控制的基本知识。这些研究将指导CAVD中抗BMP2疗法的未来测试。该建议探讨了研究的这些特定领域：CAVD和Bicuspid主动脉瓣的遗传学，基础设施（改善动物模型），细胞生物学（信号途径，机制和钙化调节）和诊断。（抽象的结尾）