Functional analysis of a novel apoptosis-inducing gene upregulated in atherosclerotic lesions

动脉粥样硬化病变中上调的新型凋亡诱导基因的功能分析

• 批准号: 15603003
• 负责人: FUKUO Keisuke
• 金额: $ 2.3万
• 依托单位: Mukogawa Women's University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15603003/
• 关键词: Apoptosis; Mitochondria; Gene; Cytochrome C; Atherosclerosis; 遺伝子解析; SNP; 疾患感受性遺伝子

Apoptosis, a programmed cell death, regulates many physiological homeostasis. Its deregulation is known to link to many human diseases such as cancer, neurodegenerative disorders, and atherosclerosis (Science 258:468,1992). There are two major pathways for induction of apoptosis. One is the extrinsic or receptor-mediated pathway represented by Fas and tumor necrosis factor α receptor. The other is the intrinsic pathway, in which mitochondria play a crucial role by releasing cytochrome C from the inter-membrane space into the cytoplasm. In this study, we have identified a novel gene upregulated in atherosclerotic lesions in apolipoprotein E-deficient mice, which are a model of atherosclerosis caused by hyperlipidemia. We compared the gene expression profile of vascular smooth muscle cells(VSMC) cultured from atherosclerotic plaque (P) to that from non-plaque in these mice. We succeeded to clone this novel gene and designated it as Apop-1. In order to clarify the function of this gene, A … More pop-1 expression vector, pcDNA3Apop-1, was introduced into cultured VSMC. Interestingly, introduction of this gene induces apoptosis of VSMC by releasing cytochrome C from mitochondria. The conditioned medium of the transfected cells had no capability to induce apoptosis, indicating that Apop-1-induced apoptosis is not mediated through secretary factors. We also demonstrated that Apop-1 gene has a mitochondria localization signal and Apop-1 protein is actually localized in mitochondria in Apop-1-transfected cells. Bcl-2 family proteins are known to regulate the release of cytochrome C from mitochondria. Importantly unlike Bcl-2 family members, Apop-1 possesses neither Bcl-2 homology (BH) nor carboxy terminal transmembrane (TM) domains. In addition, overexpression of Bcl-2 did not suppress Apop-1-induced apoptosis, suggesting that Apop-1 is a novel pro-apoptotic protein that release cytochrome C from mitochondria. We are now generating Apop-1 knockout mice and an adenoviral gene transfection system to analyze the function of this novel gene. Less

细胞凋亡是一种程序性细胞死亡，调节着许多生理稳态。已知其失调与许多人类疾病如癌症、神经变性疾病和动脉粥样硬化有关（Science 258：468，1992）。诱导细胞凋亡有两种主要途径。一种是以Fas和肿瘤坏死因子α受体为代表的外源性或受体介导的途径。另一种是内源性途径，其中线粒体通过将细胞色素C从膜间隙释放到细胞质中发挥关键作用。在这项研究中，我们已经确定了一个新的基因上调，在动脉粥样硬化病变的载脂蛋白E缺陷小鼠，这是一个模型，动脉粥样硬化引起的高脂血症。我们比较了这些小鼠动脉粥样硬化斑块（P）和非斑块培养的血管平滑肌细胞（VSMC）的基因表达谱。我们成功克隆了这个新基因，并将其命名为Apop-1。为了阐明该基因的功能，A ...更多信息 将pop-1表达载体pcDNA 3Apop-1导入培养的VSMC中。有趣的是，引入该基因通过从线粒体释放细胞色素C诱导VSMC凋亡。转染细胞的条件培养基没有诱导凋亡的能力，表明Apop-1诱导的凋亡不是通过分泌因子介导的。我们还证明了Apop-1基因具有线粒体定位信号，并且Apop-1蛋白在Apop-1转染的细胞中实际上定位于线粒体。已知Bcl-2家族蛋白调节细胞色素C从线粒体的释放。重要的是，与Bcl-2家族成员不同，Apop-1既不具有Bcl-2同源性（BH），也不具有羧基末端跨膜（TM）结构域。此外，Bcl-2的过表达不抑制Apop-1诱导的细胞凋亡，表明Apop-1是一种新的促凋亡蛋白，从线粒体释放细胞色素C。我们现在正在产生Apop-1敲除小鼠和腺病毒基因转染系统，以分析这种新基因的功能。少

项目成果

Takemura Y, Fukuo K, et al.: "Fas signaling induces Akt activation and upregulation of endothelial nitric oxide synthase expression"Hypertension. (in press). (2004)

Takemura Y、Fukuo K 等人：“Fas 信号传导诱导 Akt 激活并上调内皮一氧化氮合酶表达”高血压。

Increased plasma levels of the soluble form of Fas ligand in patients with acute myocardial infarction and unstable angina pectoris.

• DOI: 10.1016/s0735-1097(01)01800-9
• 发表时间: 2002-02
• 期刊: Journal of the American College of Cardiology
• 影响因子: 24
• 作者: M. Shimizu;K. Fukuo;S. Nagata;T. Suhara;M. Okuro;K. Fujii;Y. Higashino;M. Mogi;Y. Hatanaka;T. Ogihara

Fas Signaling Induces Akt Activation and Upregulation of Endothelial Nitric Oxide Synthase Expression

• DOI: 10.1161/01.hyp.0000120124.27641.03
• 发表时间: 2004-04
• 期刊: Hypertension: Journal of the American Heart Association
• 作者: Y. Takemura;K. Fukuo;O. Yasuda;Takahito Inoue;N. Inomata;T. Yokoi;H. Kawamoto;T. Suhara;T. Ogihara

Fukuo K, et al.: "Nifedipine upregulates manganese superoxide dismutase expression in vascular smooth muscle via endothelial cell-dependent pathways"Hypertension Research. 26. (2003)

Fukuo K 等人：“硝苯地平通过内皮细胞依赖性途径上调血管平滑肌中锰超氧化物歧化酶的表达”高血压研究。

Nifedipine indirectly upregulates superoxide dismutase expression in endothelial cells via vascular smooth muscle cell-dependent pathways

• DOI: 10.1161/01.cir.0000021924.02006.ba
• 发表时间: 2002-07-16
• 期刊: CIRCULATION
• 影响因子: 37.8
• 作者: Fukuo, K;Yang, J;Ogihara, T
• 通讯作者: Ogihara, T