COBRE: WI HOSP OF RI: P57KIP2 IN VENTRICULAR CARDIOMYOCYTE DIFFERENTIATION

COBRE：RI 的 WI HOSP：心室心肌细胞分化中的 P57KIP2

• 批准号: 7610523
• 负责人: Lazaros K. Kochilas
• 金额: $ 21.98万
• 依托单位: WOMEN AND INFANTS HOSPITAL-RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7610523
• 关键词: Animal Model; CDKN1C gene; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cell Cycle; Complementary DNA; Computer Retrieval of Information on Scientific Projects Database; Depth; Dilated Cardiomyopathy; Equilibrium; Foundations; Funding; Generations; Goals; Grant; Heart; Homologous Gene; In Situ Hybridization; Institution; Length; Mediating; Modeling; Mus; Muscle Cells; Myocardium; Natural regeneration; Pattern; Phenotype; Play; Range; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Role; Source; Testing; United States National Institutes of Health; Ventricular; Withdrawal; Zebrafish; inhibitor/antagonist; myosin light chain 2; promoter; research study; ventricular hypertrophy

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This proposal focuses on the role of the cell cycle inhibitor p57KIP2, on the differentiation of ventricular myocytes. Previous studies suggest that p57KIP2 plays an important role regulating the balance between proliferation and differentiation in the developing heart and suggests its involvement in causing a thin-walled ventricular myocardium phenotype (dilated cardiomyopathy) in the mouse. We hypothesize that cardiac over-expression of p57KIP2 depletes the ventricular proliferative zone and leads to a thin myocardium phenotype by causing early terminal differentiation of cardiomyocytes. The goal of this proposal is to test this hypothesis by examining the effects of p57 KIP2 over-expression in the mouse and zebrafish animal models. I will pursue two specific aims: 1. Examine the role of p57KIP2 in the generation of the thin-walled myocardium phenotype in the mouse. - First, I will analyze the pattern of expression of p57 KIP2 in established murine models of thin myocardium. -Second, I will examine the effects of p57 KIP2 over-expression in the mouse heart by inducing Cre-loxP mediated activation driven by the myosin light chain-2 ventricular (MLC-2v) promoter. 2. Identify the zebrafish p57 KIP2 homologue, isolate its full length cDNA and perform in depth analysis of this zebrafish homologue, including: -detailed analysis of its temporal and spatial expression pattern by whole mount in situ hybridization and RT-PCR. -study the effects of the constitutive and cardiac specific over-expression of p57 KIP2 in the zebrafish. -study the effects of the morpholino induced inactivation of p57KIP2 in the zebrafish. These experiments will form the foundation for further investigating the role of P57KIP2 in the settings of dilated cardiomyopathy, ventricular hypertrophy and cardiac regeneration. Understanding the mechanisms underlying withdrawal of cardiomyocytes from the cell cycle will be important for the treatment of a wide range of cardiovascular diseases.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 该提案重点关注细胞周期抑制剂 p57KIP2 对心室肌细胞分化的作用。先前的研究表明，p57KIP2 在调节发育中心脏的增殖和分化之间的平衡方面发挥着重要作用，并表明它参与引起小鼠薄壁心室心肌表型（扩张型心肌病）。我们假设 p57KIP2 的心脏过度表达会耗尽心室增殖区，并通过引起心肌细胞的早期终末分化而导致薄心肌表型。该提案的目的是通过检查小鼠和斑马鱼动物模型中 p57 KIP2 过度表达的影响来检验这一假设。我将追求两个具体目标： 1. 检查 p57KIP2 在小鼠薄壁心肌表型生成中的作用。 - 首先，我将分析 p57 KIP2 在已建立的薄心肌小鼠模型中的表达模式。 -其次，我将通过诱导由肌球蛋白轻链 2 心室 (MLC-2v) 启动子驱动的 Cre-loxP 介导的激活来检查 p57 KIP2 在小鼠心脏中过度表达的影响。 2. 鉴定斑马鱼p57 KIP2同源物，分离其全长cDNA并对该斑马鱼同源物进行深入分析，包括： -通过整体原位杂交和RT-PCR对其时间和空间表达模式进行详细分析。 -研究斑马鱼中 p57 KIP2 的组成型和心脏特异性过表达的影响。 -研究吗啉代诱导斑马鱼 p57KIP2 失活的影响。这些实验将为进一步研究其作用奠定基础 P57KIP2 在扩张型心肌病、心室肥厚和心脏再生中的作用。了解心肌细胞退出细胞周期的机制对于治疗多种心血管疾病非常重要。