Moga Michael-Alice, Nakamura Tomoki, Robbins Jeffrey
Department of Pediatrics, MLC7020, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229-3039, USA.
J Mol Cell Cardiol. 2008 Aug;45(2):148-55. doi: 10.1016/j.yjmcc.2008.06.002. Epub 2008 Jun 14.
The genetic, biochemical and molecular bases of human cardiac disease have been the focus of extensive research efforts for many years. Early animal models of cardiovascular disease used pharmacologic or surgical interventions, or took advantage of naturally occurring genetic abnormalities and the data obtained were largely correlative. The inability to directly alter an organism's genetic makeup and cellular protein content and accurately measure the results of that manipulation precluded rigorous examination of true cause-effect and structure-function relationships. Directed genetic manipulation in the mouse gave researchers the ability to modify and control the mammalian heart's protein content, resulting in the rational design of models that could provide critical links between the mutated or absent protein and disease. Two techniques that have proven particularly useful are transgenesis, which involves the random insertion of ectopic genetic material of interest into a "host" genome, and gene targeting, which utilizes homologous recombination at a pre-selected locus. Initially, transgenesis and gene targeting were used to examine systemic loss-of-function and gain-of-function, respectively, but further refinements in both techniques have allowed for investigations of organ-specific, cell type-specific, developmental stage-sensitive and dose-dependent effects. Genetically engineered animal models of pediatric and adult cardiac disease have proven that, when used appropriately, these tools have the power to extend mere observation to the establishment of true causative proof. We illustrate the power of the general approach by showing how genetically engineered mouse models can define the precise signaling pathways that are affected by the gain-of-function mutation that underlies Noonan syndrome. Increasingly precise and modifiable animal models of human cardiac disease will allow researchers to determine not only pathogenesis, but also guide treatment and the development of novel therapies.
多年来,人类心脏病的遗传、生化和分子基础一直是广泛研究工作的重点。早期的心血管疾病动物模型采用药物或手术干预,或利用自然发生的基因异常,所获得的数据大多具有相关性。由于无法直接改变生物体的基因组成和细胞蛋白质含量,也无法准确测量这种操作的结果,因此无法对真正的因果关系和结构 - 功能关系进行严格检验。小鼠中的定向基因操作使研究人员能够修改和控制哺乳动物心脏的蛋白质含量,从而合理设计出能够在突变或缺失的蛋白质与疾病之间提供关键联系的模型。两种已被证明特别有用的技术是转基因技术,即将感兴趣的异位遗传物质随机插入“宿主”基因组,以及基因打靶技术,该技术利用在预先选择的位点进行同源重组。最初,转基因技术和基因打靶技术分别用于研究全身性功能丧失和功能获得,但这两种技术的进一步改进使得能够研究器官特异性、细胞类型特异性、发育阶段敏感性和剂量依赖性效应。小儿和成人心脏病的基因工程动物模型已经证明,适当地使用这些工具能够将单纯的观察扩展到建立真正的因果证据。我们通过展示基因工程小鼠模型如何定义受努南综合征基础功能获得性突变影响的精确信号通路,来说明这种一般方法的作用。越来越精确且可修改的人类心脏病动物模型将使研究人员不仅能够确定发病机制,还能指导治疗和新疗法的开发。