Control of Cardiac Growth by Ca2+Dependent Phosphorylation of Histones

通过组蛋白 Ca2+ 依赖性磷酸化控制心脏生长

基本信息

批准号：
7875526
负责人：
MARK ALAN SUSSMAN
金额：
$ 18.69万
依托单位：
SAN DIEGO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-15 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7875526
关键词：
Adenoviruses Adriamycin PFS Agonist Alanine Amino Acids Apoptosis Area Biochemical Biological Assay Ca(2+)-Calmodulin Dependent Protein Kinase Calcium Calcium Signaling Calcium/calmodulin-dependent protein kinase Cardiac Cardiac Myocytes Cardiomyopathies Cell Nucleus Cells Chromatin Chromatin Structure Confocal Microscopy Couples Coupling Doxorubicin Enzymes Eukaryotic Cell Event Gene Expression Genes Genetic Transcription Goals Growth Growth Factor Heart Heart Diseases Heart Hypertrophy Heart failure Histone Code Histone H3 Histones Human Biology In Vitro Intervention Investigation Lead Link Mass Spectrum Analysis Measures Mediating Mitosis Modeling Muscle Contraction Mutate Myocardium Nuclear Organ Pathogenesis Pathology Phospho-Specific Antibodies Phosphorylation Phosphorylation Site Physiological Physiological Processes Play Post-Translational Protein Processing Protamine Kinase Protein Isoforms Protein-Serine-Threonine Kinases RNA Interference Recombinants Regulation Role Seminal Serine Signal Transduction Site-Directed Mutagenesis Small Interfering RNA Stimulus Tail Testing Time Transcriptional Regulation Transducers Ventricular Remodeling calmodulin-dependent protein kinase II cell growth chromatin immunoprecipitation chromatin remodeling design fetal insight mutant neoplastic novel novel therapeutic intervention novel therapeutics overexpression promoter public health relevance reconstitution research study response

项目摘要

DESCRIPTION (provided by applicant): Calcium is central in the regulation of many physiological processes. In the heart, calcium signals are decoded by calcium/calmodulin-dependent protein kinases (CaMKs) that regulate muscle contraction, gene expression, cell growth and apoptosis. Understanding how calcium signals are transmitted to the nucleus to modulate various cellular activities remains an important question in human biology. We have recently made a novel discovery showing that CaMKII couples calcium signals to chromatin remodeling in cardiac cells. Preliminary results show that a specific nuclear isoform of CaMKII selectively phosphorylates histone H3 in vitro and in primary cardiomyocytes. Activation of CaMK activity by agonist treatment, which induces hypertrophic growth, increases histone H3 phosphorylation. Conversely specific elimination of nuclear CaMKII in cardiac cells significantly reduces histone H3 phosphorylation and inhibits cardiac growth. These exciting new results reveal for the first time that CaMK enzymes are linked to chromatin and lead to our current hypothesis that CaMK- dependent phosphorylation of histones controls cardiac growth and may play an important role in heart pathogenesis. The goal of this application is to demonstrate this novel function of CaMK in the heart. For this, we propose to test the following hypotheses derived from our preliminary results: 1) To demonstrate the functional role of CaMK-mediated chromatin remodeling in cardiac diseases. For this we will test whether changes in histone H3 phosphorylation is detected at fetal cardiac genes and whether in vitro transcription is altered from chromatin reconstituted with mutant histones. 2) To test whether histone H3 phosphorylation is implicated in other cardiac disorders, and to begin to elucidate the biochemical mechanisms of CaMK- mediated chromatin remodeling by investigating the various amino acids that are phosphorylated by nuclear CaMKII in histone H3. The comprehensive experimental approaches will include chromatin immunoprecipitation, manipulation of gene expression using recombinant adenovirus and RNA interference in primary cardiac cells, confocal microscopy, mass spectrometry and in vitro transcription assays. This study may lead to a new area of investigation on CaMKs in the cardiac field and possibly in other fields where CaMKs exert their functional role. Most importantly, it will lead to potential new therapautic strategies to treate cardiac diseases such as heart failure. PUBLIC HEALTH RELEVANCE: Calcium/calmodulin-dependent protein kinases (CaMKs) are enzymes that are present in all major organs and are involved in a variety of cardiac disorders. We made a new discovery showing that a specific isoform of CaMK enriched in cardiac nuclei, couples calcium signals to chromatin remodeling. The goal of this application is to demonstrate that this new function of CaMK is central to pathological events leading to cardiac hypertrophy and to other cardiac diseases. Thus, the proposed study has potential to open new avenues in the CaMK field and may bring novel therapeutic intervention for the treatment of heart disease.

描述（由申请人提供）：钙在许多生理过程的调节中是核心。在心脏中，钙信号由调节肌肉收缩，基因表达，细胞生长和凋亡的钙/钙调蛋白依赖性蛋白激酶（CAMK）解码。了解如何将钙信号传输到细胞核调节各种细胞活性仍然是人类生物学的重要问题。我们最近做出了一个新的发现，表明Camkii伴侣钙信号与心脏细胞中的染色质重塑。初步结果表明，CAMKII的特定核同工型在体外和原发性心肌细胞中选择性地磷酸化了组蛋白H3。通过诱导肥厚性生长的激动剂治疗激活CAMK活性，会增加组蛋白H3磷酸化。相反，特异性消除心脏细胞中的核CAMKII可显着降低组蛋白H3磷酸化并抑制心脏生长。这些令人兴奋的新结果首次揭示了CAMK酶与染色质联系起来，并导致我们目前的假设，即组蛋白的CAMK依赖性磷酸化控制心脏生长，并且可能在心脏发病机理中起重要作用。该应用的目的是证明CAMK在心脏中的新功能。为此，我们建议测试从我们的初步结果中得出的以下假设：1）证明CAMK介导的染色质重塑在心脏疾病中的功能作用。为此，我们将测试在胎儿心脏基因上是否检测到组蛋白H3磷酸化的变化，以及是否从与突变体组蛋白重构的染色质中改变体外转录。 2）测试组蛋白H3磷酸化是否与其他心脏病有关，并开始阐明CAMK-介导的染色质重塑的生化机制，通过研究组蛋白H3中的核Camkii磷酸化的各种磷酸化。全面的实验方法将包括染色质免疫沉淀，使用重组腺病毒对基因表达的操纵以及原代心脏细胞中的RNA干扰，共聚焦显微镜，质谱和体外转录测定法。这项研究可能导致对心脏场中CAMK的新研究领域，并可能在CAMK发挥其功能作用的其他领域。最重要的是，这将导致潜在的新治疗策略来治疗心脏衰竭等心脏病。公共卫生相关性：钙/钙调蛋白依赖性蛋白激酶（CAMK）是所有主要器官中都存在的酶，并参与多种心脏疾病。我们提出了一个新的发现，表明CAMK的特定同工型富含心脏核，伴侣钙信号与染色质重塑。该应用的目的是证明CAMK的这种新功能对于导致心脏肥大和其他心脏病的病理事件至关重要。因此，拟议的研究有可能在CAMK领域开放新途径，并可能为治疗心脏病的治疗干预带来新的治疗性干预措施。