Novel Transgenic Rabbit Model for Cardiovascular Disease

描述(申请人提供)：心血管疾病(CVD)是美国最常见的单一死亡原因。虽然心血管疾病的研究大大得益于基因靶向转基因小鼠模型的使用，但这些小型啮齿动物并不能准确反映人类的心血管生理。考虑到兔在心血管解剖学、生理学和脂代谢方面与人类有许多相似之处，它将是研究心血管疾病的优秀动物模型。 Evergen的长期目标是建立一个设施，为生物医学和制药界的研究人员提供基因靶向转基因兔子。在这个项目中，Evergen BioTechnologies，Inc.建议使用一种方法，将专有的体细胞核移植和基因打靶技术相结合，生产出第一只基因靶向的转基因兔，作为CVD和其他人类疾病模型，用于研究疾病机制和开发治疗方法。 在本项目中，我们提出了培育胆固醇酯转移蛋白(CETP)基因敲除兔。到目前为止，包括辉瑞、默克、罗氏和日本烟草在内的几家公司已经开发出针对人类CETP的药物。辉瑞公司CETP抑制剂torcetRapib最近的临床试验结果不利，促使该领域的研究人员提供更好的动物模型系统，以了解CETP的功能和靶向药物对CETP的影响。 在第一阶段，我们建议在兔成纤维细胞中靶向CETP基因并产生CETP基因敲除细胞(CETP)。然后，我们将使用这些细胞进行核移植，以产生CETP KO(CETP)胚胎。 在第二阶段，我们将首先关注CETP基因敲除(CETP)兔的生产。我们将用肝脂酶转基因兔饲养CETP KO兔，以产生人源化的CETP KO兔。正常兔的肝脂酶活性明显低于人。最后对人源化CETP KO兔进行病理学评价。我们相信，通过拟议的项目，将开发出一种可行的方法来生产基因靶向转基因兔，我们将首次向世界各地的研究人员提供基因敲除兔模型。

DESCRIPTION (provided by applicant): Cardiovascular disease (CVD) is the single most common cause of death in the United States. Although the study of the CVD has greatly benefited from the use of gene-targeted transgenic mouse models, these small rodents do not accurately reflect human cardiovascular physiology. The Rabbit would be an excellent animal model for CVD research, considering its many similarities to humans in cardiovascular anatomy, physiology, and lipid metabolism. The long term goal of Evergen is to establish a facility that provides researchers in the biomedical and pharmaceutical community with gene targeted transgenic rabbits. In this project, Evergen Biotechnologies, Inc. proposes to use an approach by combining the proprietary somatic cell nuclear transfer and the gene-targeting technologies to produce the first gene-targeted transgenic rabbits as CVD and other human disease models for the study of the disease mechanisms and development of therapies. In this project, we propose to produce Cholesterol Ester Transfer Protein (CETP) knockout rabbits. So far, several companies including Pfizer, Merck, Roche, and Japan Tobacco have developed drugs targeting on human CETP. The recent unfavorable clinical trials of Pfizer's CETP inhibitor, Torcetrapib urged researchers in this field to provide a better animal model system to understand the CETP functions and effects of drugs targeting on CETP. In Phase I, we propose to target CETP gene in rabbit fibroblast cells and produce CETP knockout cells (CETP). We will then use these cells for nuclear transfer to produce CETP KO (CETP) embryos. In Phase II, we will first focus on the production of CETP knockout (CETP) rabbits. We will breed CETP KO rabbits with Hepatic Lipase transgenic rabbits to generate humanized CETP KO rabbits. Hepatic Lipase activity is much lower in normal rabbits than in human. Finally we will perform the pathology evaluation of the humanized CETP KO rabbits. We are confident that through the proposed project, a feasible method to produce gene targeted transgenic rabbits will be developed and we will for the first time provide a knockout rabbit model to the researchers worldwide.