The Molecular Basis of Cardiac Senescence

心脏衰老的分子基础

• 批准号: 7131115
• 负责人: Kenneth R Boheler
• 金额: --
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7131115
• 关键词: aging; biopsy; cell senescence; functional /structural genomics; gene expression; genetic regulation; heart ventricle; human tissue; microarray technology; myocardium; polymerase chain reaction; serial analysis of gene expression

The Laboratory of Cardiovascular Science has a strong commitment to the study of aging myocardium. To identify gene products in heart potentially involved in aging, functional genomic analyses (cDNA microarrays and Serial Analysis of Gene Expression) have been employed to analyze mRNA from left ventricles of Fisher 344 rats with or without caloric restriction, Wistar rats, C57Bl/6 mice during the perinatal period and with aging, CBA mice treated with biopeptides implicated in gerontoprotection, and human biopsies from failing and non-failing myocardium. The aim of these projects are to determine which gene products are regulated as a function of age or disease, and then use independent methods to determine the underlying mechanisms responsible for the altered changes in gene expression. In 2002, we published a reference dataset (SAGE analysis) of the mouse myocardium that is publicly available, and we have expanded these studies to examine male versus female mice and young versus old mice. We have completed a large-scale transcriptome analysis of an aging Fisher 344 aging rat model and compared these results with other rodent strains. We are completing an extensive quantitative-PCR analysis to validate these data, and by independent techniques, to determine the significance of the changes in gene expression with the aging process. We have expanded our aging studies in rodent to include in vitro analyses of cultured cardiomyoyctes and fibroblasts to delineate mechanisms underlying the aging response of cardiac genes, and are in the process of elucidating genetic correlates underlying these changes in transcript abundance. Finally, we have employed functional genomic techniques (microarrays) to examine the transcriptomes of LVs from failing (n=8) and non-failing human myocardium (n=7). Following identification of a pool of HF-responsive candidate genes by microarrays and statistical methods, we employed Q-PCR on a larger sample population (n=34) to validate and examine the role of contributing biological variables (age and gender). We find that most of the HF-candidate genes (including transcription factors, modifying enzymes, ECM proteins and metabolic enzymes) demonstrated significant changes in gene expression; however, the majority of the putative changes depended on variables such as sex and age, and not on HF alone. Additionally, some putative HF-responsive gene products demonstrated highly significant changes in expression as a function of age and/or sex, but independent of HF. These studies with human samples have now expanded to include hypertension, and we have begun a meta analysis of human heart failure data. The data from each of these projects are being compared in an effort to identify and analyze the function of candidate genes in aging and disease.

心血管科学实验室致力于衰老心肌的研究。为了鉴定心脏中可能与衰老相关的基因产物，已采用功能基因组分析（cDNA 微阵列和基因表达系列分析）来分析来自有或没有热量限制的 Fisher 344 大鼠、Wistar 大鼠、C57Bl/6 小鼠左心室的 mRNA在围产期和随着年龄的增长，CBA 小鼠接受了与老年保护有关的生物肽治疗，并对衰竭和非衰竭心肌进行了人体活检。这些项目的目的是确定哪些基因产物作为年龄或疾病的函数受到调节，然后使用独立的方法来确定导致基因表达改变的潜在机制。 2002 年，我们发布了公开的小鼠心肌参考数据集（SAGE 分析），并且我们将这些研究扩展到检查雄性小鼠与雌性小鼠以及年轻小鼠与老年小鼠。 我们已经完成了对老化 Fisher 344 衰老大鼠模型的大规模转录组分析，并将这些结果与其他啮齿动物品系进行了比较。我们正在完成广泛的定量 PCR 分析来验证这些数据，并通过独立技术来确定基因表达随衰老过程变化的重要性。我们已经扩大了啮齿类动物的衰老研究，包括对培养的心肌细胞和成纤维细胞的体外分析，以描绘心脏基因衰老反应的机制，并且正在阐明转录丰度这些变化背后的遗传相关性。 最后，我们采用功能基因组技术（微阵列）来检查失败（n = 8）和非失败人类心肌（n = 7）的左心室的转录组。通过微阵列和统计方法鉴定出一系列 HF 反应候选基因后，我们在更大的样本群体 (n=34) 上采用 Q-PCR 来验证和检查贡献生物变量（年龄和性别）的作用。我们发现大多数 HF 候选基因（包括转录因子、修饰酶、ECM 蛋白和代谢酶）在基因表达上表现出显着的变化；然而，大多数假定的变化取决于性别和年龄等变量，而不仅仅取决于心力衰竭。此外，一些假定的心力衰竭反应基因产物表现出高度显着的表达变化，作为年龄和/或性别的函数，但与心力衰竭无关。这些针对人类样本的研究现已扩展到包括高血压，我们已经开始对人类心力衰竭数据进行荟萃分析。正在对每个项目的数据进行比较，以识别和分析候选基因在衰老和疾病中的功能。