MCP-1 induced gene expression in cardiovascular disease

MCP-1 诱导心血管疾病基因表达

• 批准号: 6769538
• 负责人: PAPPACHAN KOLATTUKUDY KOLATTUKUDY
• 金额: $ 36万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-20 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6769538
• 关键词: apoptosis; cardiac myocytes; cytokine; flow cytometry; gene expression; gene induction /repression; genetically modified animals; heart ventricle; immunocytochemistry; laboratory mouse; microarray technology; monocyte; monocyte chemoattractant protein 1; myocardial ischemia /hypoxia; myocardium; polymerase chain reaction; terminal nick end labeling; tissue /cell culture

DESCRIPTION (provided by applicant): Many recent reports provide strong evidence that MCP- 1 plays a major role in the development of ischemic heart disease (IHD) that is responsible for the majority of the 5 million human heart failure cases in the US, but the underlying mechanism is not known. We postulate that MCP-1-induced gene expression changes in the monocytes and the consequent production of cytokines and other biologically active molecules, as well as possible direct effects of MCP- 1 on the major cell types in the myocardium can lead to the development of IHD. IHD probably develops as a net consequence of the interplay of these events which involve many cell types and many biologically active molecules. Therefore in vitro studies on isolated cells cannot accurately reflect the interactions involved in IND. On the other hand a genomic approach can discover the genes whose altered expression in the heart leads to the development of IHD. Such an approach, although not possible on humans, can be done on a suitable animal model. We have developed a transgenic murine model that faithfully recapitulates most features of human IRD. Use of this model has a high likelihood of discovering novel genes as illustrated by our discovery of a previously unknown MCP-1-induced protein (MCPIP). With this overall objective we propose to pursue the following specific aims: 1.) Identify gene expression changes that occur in the ventricle during IHD development induced by targeted expression of MCP 1 in the CMC of transgenic mice using gene chip technology. 2.) Determine the function of the novel MCPIP: a) Determine whether a) MCPIP expression causes cell death, b) this death is enhanced by MCP-l and its receptor CCR2, c) whether the cell death shares features characteristic of apoptosis, d) MCPIP has a transcription factor-like activity and whether structural features required for this activity are also required for the cell death-inducing activity. 3.) Assess the role of apoptosis in the development of MCP 1-induced cardiovascular disease by testing whether a) prevention of monocyte apoptosis by targeted expression of Bc12 or b) prevention of apoptosis by absence of Fas or expression of sFas in the myocytes would delay or rescue the development of the disease. The approach proposed here will identify MCP-l induced alterations in the expression of known genes not previously implicated in IHD and novel genes of unknown function that are involved in IHD. These genes are likely to reveal novel targets for intervention in IND and genes with prognostic value.

描述（由申请人提供）：许多最近的报告提供了强有力的证据，表明 MCP-1 在缺血性心脏病 (IHD) 的发展中发挥着重要作用，而缺血性心脏病 (IHD) 是美国 500 万人类心力衰竭病例中的大多数的原因，但其根本机制尚不清楚。我们假设 MCP-1 诱导的单核细胞基因表达变化以及随后产生的细胞因子和其他生物活性分子，以及 MCP-1 对心肌主要细胞类型可能的直接影响可能导致 IHD 的发生。 IHD 的发生可能是这些事件相互作用的最终结果，涉及多种细胞类型和多种生物活性分子。因此，对分离细胞的体外研究无法准确反映 IND 中涉及的相互作用。另一方面，基因组方法可以发现心脏中表达改变导致 IHD 发生的基因。这种方法虽然不可能在人类身上实现，但可以在合适的动物模型上实现。我们开发了一种转基因小鼠模型，它忠实地再现了人类 IRD 的大部分特征。使用该模型很有可能发现新基因，正如我们发现以前未知的 MCP-1 诱导蛋白 (MCPIP) 所示。在此总体目标下，我们建议追求以下具体目标： 1.) 使用基因芯片技术鉴定转基因小鼠 CMC 中 MCP 1 的靶向表达诱导的 IHD 发育过程中心室中发生的基因表达变化。 2.)确定新型MCPIP的功能：a)确定a)MCPIP表达是否引起细胞死亡，b)这种死亡被MCP-1及其受体CCR2增强，c)细胞死亡是否具有细胞凋亡的特征，d)MCPIP具有转录因子样活性以及该活性所需的结构特征是否也是细胞死亡诱导活性所需要的。 3.) 通过测试 a) 通过 Bc12 的靶向表达预防单核细胞凋亡或 b) 通过肌细胞中缺乏 Fas 或 sFas 表达来预防细胞凋亡是否会延迟或挽救疾病的发展，评估细胞凋亡在 MCP 1 诱导的心血管疾病发展中的作用。这里提出的方法将鉴定MCP-1诱导的先前未涉及IHD的已知基因的表达改变以及涉及IHD的功能未知的新基因。这些基因可能会揭示 IND 干预的新靶点和具有预后价值的基因。