Basic Mechanisms of Human Calcific Aortic Valve Disease

人类钙化性主动脉瓣疾病的基本机制

• 批准号: 8535815
• 负责人: Lauren D. Black III
• 金额: $ 37.39万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-23 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8535815
• 关键词: AXIN1 gene; Adverse drug effect; Affect; Animals; Awareness; Bicuspid; Biological; Biological Testing; Biology; Blood; CDH13 gene; Cadherins; Calcified; Calcium; Candidate Disease Gene; Cell Communication; Cells; Characteristics; Congenital Heart Defects; Cyclin-Dependent Kinases; Data Set; Deposition; Development; Disease; Echocardiography; Elderly; Endoglin; Engineering; Extracellular Matrix; Framingham Heart Study; Gene Expression Profile; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Growth; Heart; Heart Valve Diseases; Human; Human Genetics; Life; Measures; Medical; Modeling; Mus; Mutation; Nucleotides; Operative Surgical Procedures; Osteogenesis; Pathway Analysis; Pathway interactions; Patients; Phenotype; Phosphorylation; Physiologic calcification; Population; Property; Qualifying; RNA Splicing; Regulation; Research; Retinoblastoma Protein; Role; Signal Pathway; Signal Transduction; Single Nucleotide Polymorphism; Stem cells; Stenosis; Stimulus; Surveys; Testing; Therapeutic; Tissues; Transcript; Translating; Variant; Work; abstracting; aortic valve; aortic valve disorder; bone; calcification; cell growth regulation; disease phenotype; experience; gain of function; genetic analysis; genome wide association study; genome-wide; insight; interstitial cell; loss of function mutation; mineralization; mouse model; notch protein; osteogenic; osteoprogenitor cell; prevent; programs; promoter; response; soft tissue; transcription factor; transcriptome sequencing; valve replacement

DESCRIPTION (provided by applicant): Progressive calcific aortic valve disease (CAVD) is characterized by calcium deposition in the valve cusps as well as frank bone formation. People born with a bicuspid aortic valve (BAV), a common congenital heart defect, can develop significant CAVD during the prime of life. BAV and CAVD are regulated by genetic factors including mutations in the Notch1 gene. We analyzed human genetic datasets to identify gene variants associated with BAV and/or CAVD. Our genetic analysis of the Tufts BAV Genetics Study identified gene variants in the WNT/b-catenin and TGFb signaling pathways, which are known to contribute to aortic valve development as well as valve calcification. Our unpublished genome-wide association analysis of the Framingham Heart Study has further identified gene variants in the cyclin dependent kinase (CDK), the soft tissue calcification and the cell-cell communication pathways associated with CAVD. The association of CDK7 gene SNPs with aortic valve calcification is provocative because we have previously demonstrated that the retinoblastoma protein (pRb) and CDK pathway regulate bone formation through the runx2 transcription factor and by regulating cellular responses to Notch1. Taken together we put forth an over-arching hypothesis that the pRb-CDK pathway regulates CAVD through transcriptional effects on soft tissue calcification and cell-cell communication genes regulated by runx2 and Notch1. We will test this hypothesis in our first specific aim by comparing pRb-deficient and control valve interstitial cell (VIC) growh and expression of bone and calcification gene markers in response to factors that stimulate mineralization. Next, we will compare pRb-deficient and control VIC extracellular matrix stiffness characteristics and calcification. Finally we will determine whether pRb regulates runx2 occupancy of soft tissue calcification gene promoters in VICs. In our second aim we will study mice engineered to have a loss- or a gain-of-function in CDK6 that have significant bone and mineralization phenotypes. Further, we will define the role of cdk6 in regulation of VIC osteogenic responses to Notch, and we will survey pRb-cdk mouse models for aortic valve calcification. In our third aim we will analyze expression and activity of pRb-cdk components in human calcified aortic valve tissue. Finally, to broadly survey expression of the pRb-cdk pathway, as well as genes that regulate soft-tissue calcification and cell-cell communication, we will sequence the CAVD transcriptome (RNA-seq). We have assembled a team of experts with experience in CAVD genetics, the regulation of the osteogenic transcriptional program and calcification by the pRb-CDK pathway, and the heart extracellular matrix. We anticipate that our studies will provide unique insights to the biology of CAVD and that this work will translate meaningfully to the understanding of calcific aortic valve disease as well as help guide potential therapeutic options or raise awareness of potential adverse drug effects on the aortic valve. (End of Abstract)

描述（由申请人提供）： 进行性钙化性主动脉瓣疾病（CAVD）的特征是瓣尖钙沉积以及明显的骨形成。出生时带有二叶式主动脉瓣（BAV）的人，是一种常见的先天性心脏病，在生命的黄金时期可能会发生严重的CAVD。BAV和CAVD受包括Notch 1基因突变在内的遗传因素调节。我们分析了人类遗传数据集，以确定与BAV和/或CAVD相关的基因变异。我们对Tufts BAV遗传学研究的遗传分析确定了WNT/b-连环蛋白和TGF β信号通路中的基因变异，已知这些基因变异有助于主动脉瓣发育以及瓣膜钙化。我们未发表的全基因组关联分析的心脏研究进一步确定了基因变异的细胞周期蛋白依赖性激酶（CDK），软组织钙化和细胞间通讯途径与CAVD。CDK 7基因SNPs与主动脉瓣钙化的相关性是具有挑衅性的，因为我们先前已经证明视网膜母细胞瘤蛋白（pRb）和CDK通路通过runx 2转录因子和调节细胞对Notch 1的反应来调节骨形成。综上所述，我们提出了一个过度假设，即pRb-CDK通路通过对软组织钙化和细胞间通讯基因的转录作用调节CAVD，这些基因由runx 2和Notch 1调节。我们将在我们的第一个具体目标中通过比较pRb缺陷和对照瓣膜间质细胞（维克）生长以及骨和钙化基因标记物对刺激矿化的因素的反应来检验这一假设。接下来，我们将比较pRb缺陷和对照维克细胞外基质硬度特征和钙化。最后，我们将确定是否pRb调节软组织钙化基因启动子runx 2占用VIC。在我们的第二个目标中，我们将研究具有显著骨和矿化表型的CDK 6功能缺失或获得的小鼠。此外，我们将确定cdk 6在调节维克对Notch的成骨反应中的作用，我们将调查pRb-cdk小鼠模型的主动脉瓣钙化。在我们的第三个目标中，我们将分析pRb-cdk组分在人钙化主动脉瓣组织中的表达和活性。最后，为了广泛调查pRb-cdk通路的表达，以及调节软组织钙化和细胞间通讯的基因，我们将对CAVD转录组（RNA-seq）进行测序。我们已经组建了一个专家团队，他们在CAVD遗传学、通过pRb-CDK途径调节成骨转录程序和钙化以及心脏细胞外基质方面具有丰富的经验。我们预计，我们的研究将为CAVD的生物学提供独特的见解，这项工作将有意义地转化为对钙化性主动脉瓣疾病的理解，并有助于指导潜在的治疗选择或提高对主动脉瓣潜在不良药物影响的认识。 (End摘要）