Dynamic Expression Profiling in the Intact Human Heart

描述（由申请人提供）： 导致进行性心肌功能障碍的机制， 心肌病性衰竭的人类心脏的重塑是未知的。在 一般来说，这些病理生理机制可能涉及改变 心肌基因表达。最近的许多研究表明， 为了有意义，基因调控和表达必须在 完整的心脏本提案的总体目标是调查， 患有扩张型心肌病引起的心肌衰竭的人类受试者 表型，基因表达谱的效用进行纵向 因为表型是动态调节的。我们提出“动态表达 分析”可以识别基因类别以及特定的个体新的 表达改变可能与表型相关的基因 改进.该提案检验了一个普遍假设， 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增加相关， 细胞骨架、细胞外基质、信号内表达减少 转导，生长因子，转录/翻译/核苷酸合成， 和细胞周期/凋亡基因类别。“提案中的三个具体目标 分别处理基因、个体 基因，以及与表型改善相关的基因变化动力学， 特发性扩张型心肌病对氧阻断剂的反应。第四个目标 比较了Affyssin基因芯片方法和 定量RT-PCR检测38个基因。我们已经开发出技术来测量 在少量的人类细胞中表达大量的靶基因， 心室心肌，可通过以下方法从完整心脏连续获得 右心室（RV）肌内膜活检，使用Affyssin基因芯片， 定量RT-QPCR。我们已经证明了RT-QPCR用于系列， 纵向方式能够识别基因，其改变的表达是 收缩功能障碍和心室/肌细胞的可能解释 重塑我们还证明了表达谱分析的实用性， 一系列纵向研究，其中表型受治疗调节，我们 提供证据证明这种方法上级“静态”表达 在横截面设计的背景下进行的仿形。

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.