Dynamic Expression Profiling in the Intact Human Heart

描述(由申请人提供)： 进展性心肌功能障碍的发病机制 心肌病、衰竭的人类心脏的重塑尚不清楚。在……里面 一般说来，这些病理生理机制可能涉及到改变 在心肌基因表达上。最近的许多研究已经证明， 为了有意义，基因的调节和表达必须在 完整的心脏。这项提案的总体目标是调查， 扩张型心肌病致心力衰竭的受试者 表型，基因表达谱的实用性是纵向执行的 因为表型是动态调节的。我们提出“动态表达” 侧写》可以识别基因类别以及特定的个体小说 表达变化与表型有潜在因果关系的基因 进步。该提案测试了一个普遍的假设，该假设得到了 初步数据：“扩张型心肌病表型的改善是 与代谢类别基因表达的增加有关，以及 细胞骨架、细胞外基质、信号的表达减少 转导、生长因子、转录/翻译/核苷酸合成， 和细胞周期/细胞凋亡基因分类。 分别处理基因、个体类别的鉴定 与表型改良相关的基因和基因变化的动力学 特发性扩张型心肌病对0-阻滞剂的反应。第四个目标 Affymetrix基因芯片方法与基因芯片定量方法的比较 定量逆转录聚合酶链式反应，38个基因。我们已经开发了一些技术来测量 大量靶基因在少量人中的表达 可通过以下方式从完整心脏连续获得的心室心肌 右室心内膜心肌活检，使用Affymetrix GeneChips和 定量RT-QPCR法。我们已经证明了RT-QPCR在一系列， 纵向时尚能够识别其改变表达的基因是 收缩功能障碍和腔/肌细胞的可能解释 改建。我们还演示了表达分析在 连续的、纵向的研究，其中表型受治疗的影响，我们 提供证据，证明这种方法优于“静态”表达 在横截面设计的背景下执行的轮廓分析。

DESCRIPTION (provided by applicant): The mechanisms responsible for progressive myocardial dysfunction and remodeling of the cardiomyopathic, failing human heart are unknown. In general, these pathophysiologic mechanisms are likely to involve alterations in myocardial gene expression. Numerous recent studies have demonstrated that, in order to be meaningful, gene regulation and expression must be examined in the intact heart. The overall objective of this proposal is to investigate, in human subjects with myocardial failure from a dilated cardiomyopathy phenotype, the utility of gene expression profiling performed longitudinally as phenotype is dynamically modulated. We propose that "dynamic expression profiling" can identify gene categories as well as specific individual novel genes whose altered expression is potentially causally related to phenotypic improvement. The proposal tests one general hypothesis supported by preliminary data: that "improvement in the dilated cardiomyopathy phenotype is associated with an increase in metabolic category gene expression, and a decrease in expression within cytoskeletal, extracellular matrix, signal transduction, growth factors, transcription /translation/nucleotide synthesis, and cell cycle/apoptosis gene categories." Three Specific Aims in the proposal deal respectively with identification of categories of genes, individual genes, and kinetics of gene changes associated with phenotypic improvement in idiopathic dilated cardiomyopathy in response to 0-blocking agents. A 4th Aim compares mRNA quantitation between the Affymetrix GeneChip method and quantitative RT-PCR, for 38 genes. We have developed techniques to measure the expression of a large number of target genes in small quantities of human ventricular myocardium that can be obtained serially from the intact heart by right ventricular (RV) endomyocardial biopsy, using Affymetrix GeneChips and quantitative RT-QPCR. We have demonstrated that RT-QPCR used in a serial, longitudinal fashion is able to identify genes whose altered expression is a potential explanation for contractile dysfunction and chamber/myocyte remodeling. We have also demonstrated the utility of expression profiling in serial, longitudinal studies where phenotype is modulated by treatment, and we provide evidence that this approach is superior to "static" expression profiling performed within the context of a cross-sectional design.