Production of Cloned Miniature Pigs Expressing High Levels of Human Apolipoprotein(a) in Plasma.
Production of Cloned Miniature Pigs Expressing High Levels of Human Apolipoprotein(a) in Plasma.
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DOI：
10.1371/journal.pone.0132155
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Yoshida M
中科院分区：
文献类型：
--
作者：
Ozawa M;Himaki T;Ookutsu S;Mizobe Y;Ogawa J;Miyoshi K;Yabuki A;Fan J;Yoshida M
高脂蛋白(a) [Lp(a)]水平是动脉粥样硬化发展的主要危险因素。然而，由于载脂蛋白(a)的独特成分载脂蛋白(a) [apo(a)]仅存在于灵长类动物和人类中，因此由于缺乏适当的动物模型，对人类Lp(a)的研究一直受到阻碍。利用体细胞核移植（SCNT）技术，我们培育了在血浆中表达人载脂蛋白(a)的转基因小型猪。首先，我们将血凝素（HA）标记的人载脂蛋白(a) cDNA置于β-肌动蛋白启动子和巨细胞病毒增强子的控制下，然后将其导入肾上皮细胞。用抗ha抗体对细胞进行免疫染色，可以鉴定稳定表达载脂蛋白的细胞(a)；其中一个阳性克隆被用来为SCNT提供供体细胞，产生表达载脂蛋白(a)的囊胚。克隆仔猪各器官组织切片的免疫组化分析和总RNA的RT-PCR分析显示，apo(a)在心脏、肝脏、肾脏和肠道等多个组织/器官中均有表达。更重要的是，转基因品系的血浆中Lp(a)含量高（约400 mg/dL），并将转基因载脂蛋白(a)基因传递给后代。因此，我们产生了一个人类载脂蛋白(a)转基因的微型猪，可以作为模型系统来研究与人类疾病相关的晚期动脉粥样硬化。猪和人类心血管系统在解剖学和生理学上的相似性将使该猪模型成为研究载脂蛋白(a)在动脉粥样硬化形成中的作用以及血管健康和疾病的潜在机制的宝贵信息来源。
High lipoprotein(a) [Lp(a)] levels are a major risk factor for the development of atherosclerosis. However, because apolipoprotein(a) [apo(a)], the unique component of Lp(a), is found only in primates and humans, the study of human Lp(a) has been hampered due to the lack of appropriate animal models. Using somatic cell nuclear transfer (SCNT) techniques, we produced transgenic miniature pigs expressing human apo(a) in the plasma. First, we placed the hemagglutinin (HA)-tagged cDNA of human apo(a) under the control of the β-actin promoter and cytomegalovirus enhancer, and then introduced this construct into kidney epithelial cells. Immunostaining of cells with anti-HA antibody allowed identification of cells stably expressing apo(a); one of the positive clones was used to provide donor cells for SCNT, yielding blastocysts that expressed apo(a). Immunohistochemical analysis of tissue sections and RT-PCR analysis of total RNA from organs of cloned piglet revealed that apo(a) is expressed in various tissues/organs including heart, liver, kidney, and intestine. More importantly, a transgenic line exhibited a high level (>400 mg/dL) of Lp(a) in plasma, and the transgenic apo(a) gene was transmitted to the offspring. Thus, we generated a human apo(a)–transgenic miniature pig that can be used as a model system to study advanced atherosclerosis related to human disease. The anatomical and physiological similarities between the swine and human cardiovascular systems will make this pig model a valuable source of information on the role of apo(a) in the formation of atherosclerosis, as well as the mechanisms underlying vascular health and disease.
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影响因子：
8.7
作者：
Fan, JL;Shimoyamada, H;Watanabe, T
通讯作者：
Watanabe, T
DOI：
10.1006/bbrc.1999.0242
发表时间：
1999-02-24
影响因子：
3.1
作者：
Fan, JL;Araki, M;Watanabe, T
通讯作者：
Watanabe, T
影响因子：
17.1
作者：
Al-Mashhadi, Rozh H.;Sorensen, Charlotte B.;Bentzon, Jacob F.
通讯作者：
Bentzon, Jacob F.
影响因子：
1.6
作者：
Himaki, Takehiro;Mori, Hironori;Yoshida, Mitsutoshi
通讯作者：
Yoshida, Mitsutoshi
影响因子：
--
作者：
Mastromonaco, Gabriela F.;D Perrault, Steve;King, W. Allan
通讯作者：
King, W. Allan